Searchable abstracts of presentations at key conferences on reproductive biology and medicine
Volume 1 | WCRB2014 | Next issue

World Congress of Reproductive Biology 2014

Edinburgh, UK
02 Sep 2014 - 04 Sep 2014

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World Congress on Reproductive Biology 2014, 02 - 04 September 2014; Edinburgh, UK

ra0001p031 | (1) | WCRB2014

Mitochondrial dynamics controlled by mitofusins direct organelle positioning and movement during meiotic division

Wakai Takuya , Harada Y , Miyado K , Kono Tomohiro

Introduction: Mitochondria are abundant in fully-grown mammalian oocytes with a unique spherical morphology, but the manner in which the behavior of mitochondria is controlled is not well understood. We explored the spatiotemporal control of mitochondrial morphology by mitochondrial fusion and fission mechanisms and the functional impact of that control on meiotic division, using mouse oocytes.Materials and methods: Immature oocytes were collected from t...

ra0001p032 | (1) | WCRB2014

The detailed localization of meiotic cohesin subunits, RAD21L and REC8, in mouse spermatocytes

Rong Mei , Matsuda Atsushi , Hiraoka Yasushi , Lee Jibak

Introduction: In meiosis, homologous chromosomes pair, synapse, and recombine with their partners in parallel with the formation of the synaptonemal complex, a tripartite structure with two axial elements connected by transverse filaments. It has been demonstrated that two meiosis-specific cohesin subunits, RAD21L and REC8 are essential for the formation of the axial elements and homologous chromosome recombination. However, it is unknown how they are involved in synapsis and ...

ra0001p033 | (1) | WCRB2014

Atrazine exposure of adult mice affect spermatogenesis

Gely-Pernot Aurore , Hao Chunxiang , Kervarec Christine , Stuparevic Igor , Chalmel Frederic , Evrard Bertrand , Priming Michael , Jegou Bernard , Smagulova Fatima

Declining human fertility has become a serious public health problem in many countries around the world. There is a rapidly growing number of evidence that human reproductive health is negatively affected by various environmental factors including life style and exposure to chemical compounds such as pesticides, phthalates, and bisphenol A (BPA). The aim of our study is to understand how commonly used pesticides can affect meiosis, the central event of mammalian gametogenesis....

ra0001p034 | (1) | WCRB2014

C-type natriuretic peptide stimulates resumption of meiosis via a cGMP-dependant mechanism in porcine oocytes

Rose Ryan D , Sugimura Satoshi , Ritter Lesley J , Brown Hannah M , Thompson Jeremy G , Gilchrist Robert B

Research conducted in recent years has led to great advances in our understanding of the participation of cGMP in meiosis. It is clear that increased intra-oocyte concentrations of cGMP inhibit meiosis in mouse models. Like cAMP, cGMP may also have a meiotic stimulatory function, possibly via cGMP/PKG. Abattoir derived gilt porcine ovaries were collected, antral follicles aspirated and oocytes collected and cultured in TCM-199+3 mg/ml BSA. Natriuretic peptide receptor 2 (N...

ra0001p035 | (1) | WCRB2014

Zinc regulate meiotic resumption and metaphase arrest in porcine oocyte

Zhao Ming-Hui , Kwon Jung-Woo , Liang Shuang , Kim Seon-Hyang , Kim Teoan , Kim Nam-Hyung , Cui Xiang-Shun

Introduction: Zinc is an extremely important trace element that play important roles in several biological processes. In this study, we investigated the role of zinc during meiotic resumption and metaphase arrest in in vitro-matured porcine oocytes.Materials and methods: Oocytes which arrest at GV or MII stage were treated with TPEN, a Zn2+ chelator, respectively. Meiotic resumption and activation were assayed. Effect of PMA, a PKC ac...

ra0001p036 | (1) | WCRB2014

Morphological markers to select populations of oocytes with different cultural needs for dedicated pre-maturation systems

Dieci Cecilia , Labrecque Remi , Lodde Valentina , Tessaro Irene , Baruffini Valentina , Lodi Giorgia , Modina Silvia , Sirard Marc-Andre , Luciano Alberto M

Introduction: Several studies support the notion that in vitro pre-maturation treatments aimed to improve the developmental capability of immature oocytes have a different effect depending on the oocyte metabolic status at the time of its removal from the follicle. We demonstrated that changes in large-scale chromatin configuration within the germinal vesicle (GV) of fully-grown bovine oocytes are related to the acquisition of developmental competence. In particular, ...

ra0001p037 | (1) | WCRB2014

Abstract withdrawn....

ra0001p038 | (1) | WCRB2014

The gametic synapse; transferring RNA to the oocyte

Macaulay Angus D , Gilbert Isabelle , Caballero Julieta , Scantland Sara , Fournier Eric , Tossou Prudencio , Sirard Marc-Andre , Clarke Hugh , Khandjian Edward , Richard Francois , Hyttel Poul , Robert Claude

Introduction: Oocytes must effectively grow, accumulate reserves, and mature prior to fertilization. Errors in any of these processes affecting nuclear, cytoplasmic, or molecular maturation of the oocyte will result in poor embryonic development.Materials and methods: Immunofluorescence, direct fluorescent staining, and autoradiography were combined with confocal, epifluorescent, and electron microscopy to study the transzonal projections (TZPs) between ...

ra0001p039 | (1) | WCRB2014

Dissecting the meiotic gene network in female embryonic germ cells

Abby Emilie , Tourpin Sophie , Guerquin Marie-Justine , Le Bouffant Ronan , Habert Rene , Livera Gabriel

Context: In mammalian female germ cells, meiotic entry occurs during fetal life. The current model places retinoic acid (RA) as a key signal inducing meiotic initiation by activating the expression of stimulated by retinoic acid 8 (Stra8), a necessary factor for pre-meiotic chromosomal replication. However, the requirement of RA for meiotic initiation has been recently questioned in the female germ cells.Material and method: To clarify whether R...

ra0001p040 | (1) | WCRB2014

The oocyte influences cumulus expansion and glucose metabolism during IVM in cattle

Lima Paula F , Price Christopher A , Buratini Jose

Oocyte–cumulus communication is essential for COC metabolism and oocyte developmental competence. Although the oocyte appears not to be absolutely required for cumulus expansion in cattle, it is not clear whether it influences this process. We tested the effects of oocytectomy on expansion and glucose metabolism of bovine COCs submitted to in vitro maturation (IVM). Follicles 3–8 mm were aspirated from abattoir ovaries and grades 1 and 2 COCs were selected. ...

ra0001p041 | (1) | WCRB2014

Plk1 plays a role in cortical actin polymerisation during meiosis I

Yuen Wai Shan , Zhang Qing-Hua , Carroll John

Female meiosis involves a highly asymmetrical division to form a large secondary oocyte and a small polar body. Polo-like kinase I (Plk1) is a serine/threonine kinase which is highly conserved from yeast to human and is a potent regulator of mitosis including cytokinesis. Plk1 is known to regulate myosin via the activation of RhoA which leads to the contraction of the cleavage furrow in mitotic cells. Indirect evidence has also shown that Plk1 may regulate Cdc42 activity throu...

ra0001p042 | (1) | WCRB2014

Cyclin A2 is essential for the chromosome segregation during the meiosis I of the mouse oocyte maturation

Zhang Qing-Hua , Yuen Wai Shan , Carroll John

Cyclin A, the first cyclin ever cloned, is thought to be an essential component of the cell-cycle engine. Mammalian cells encode two A-type cyclins, a testis-specific cyclin A1 and a ubiquitously expressed cyclin A2. Cyclin A2 is an essential mitotic CDK regulatory partner and is attributed with a wide range of effects early in the G2–M transition. Although its role in mitosis has been extensively investigated, research into the role of cyclin A2 in meiosis is lacking. In...

ra0001p043 | (1) | WCRB2014

Effect of cAMP regulators on bovine cumulus–oocyte communication and embryo development in vitro

Nogueira Marcelo G , Machado Mariana F , Li Haijun , Razza Eduardo M , Gilchrist Robert B , Sutton-McDowall Melanie L , Thompson Jeremy G

Introduction: Preventing spontaneous maturation in vitro by adding cAMP regulators is thought to maintain the communication between oocyte and cumulus cells via gap junction communication (GJC), hence promoting developmental competence. We aimed to assess the integrity of gap junction maintenance in cumulus oocyte complexes (COCs) after pre-maturation with IBMX and Forskolin (FSK) and the influence on bovine embryo development.Materials...

ra0001p044 | (1) | WCRB2014

Insight into progesterone receptor membrane component 1 action during bovine oocyte meiosis by means of siRNA-mediated gene silencing

Lodde Valentina , Tessaro Irene , Raucci Franca , Franciosi Federica , Marchese Federica , Modina Silvia , Peluso John J. , Luciano Alberto M

Introduction: Previous studies suggest that progesterone receptor membrane component 1 (PGRMC1) plays an essential role during bovine oocyte meiosis, since it i) localizes to the centromeres at metaphases I and II and ii) concentrates between the separating chromosomes at ana/telophase I. Moreover, injection of an antibody to PGRMC1 significantly impairs completion of meiosis. The aim of the present study is to expand these findings by using siRNA (RNAi)-mediated gene silencin...

ra0001p045 | (1) | WCRB2014

Maturation conditions do not affect Myst1, Hat1, and Sirt1 mRNA abundance in horse oocytes

Tessaro Irene , Franciosi Federica , Papillier Pascal , Goudet Ghylene , Dalbies-Tran Rozenn , Douet Cecile , Reigner Fabrice , Deleuze Stefan , Lodde Valentina , Luciano Alberto M.

Introduction: We recently demonstrated that in vitro maturation (IVM) is associated with defects in histone H4 lysine 16 (H4K16) acetylation in horse oocytes, together with a higher incidence of spindle anomalies and aneuploidy. In the present study we investigated whether maturation conditions can alter the abundance of transcripts involved in histone acetylation and deacetylation. The specific H4K16 acetyl-transferase Myst1, the general acetyl-transferase <...

ra0001p046 | (1) | WCRB2014

Inhibition of mTOR signaling induces cumulus expansion and stimulates meiotic maturation of oocytes in mice

Hoshino Yumi , Sato Eimei

The mammalian target of rapamycin (mTOR) signaling pathway functions as a central regulator of cell growth, proliferation, and survival. We previously reported that during meiotic maturation, the expression levels of mTOR in oocytes remain similar from the germinal vesicle (GV) stage to metaphase II (MII). To investigate the role played by mTOR during meiotic resumption, we cultured murine cumulus – oocyte complexes (COCs) in the presence of mTOR inhibitors. mTOR expressi...

ra0001p047 | (1) | WCRB2014

Inhibition of DMRTA2 impairs human female germline development in xeno-grafted ovaries

Poulain Marine , Tourpin Sophie , Muczynski Vincent , Messiaen Sebastien , Moison Delphine , Duquenne Clotilde , Benachi Alexandra , Rouiller-Fabre Virginie , Habert Rene , Frydman Nelly , Livera Gabriel

Context: DMRTA2 belongs to a family of genes coding for proteins containing a DM-domain that are conserved among vertebrates and widely involved in gonadal differentiation. We recently identified Dmrta2 gene expression through transcriptome analysis performed in murine female embryonic germ cells and retrieved its expression in human fetal ovaries. The role of DMRTA2 is poorly documented and we thus conducted this study to clarify its implication in ...

ra0001p048 | (1) | WCRB2014

Abnormal behavior of lysine acetylation during one-cell stage mouse cloned embryos

Kishigami Satoshi , Reum Lee Ah , Ha Le Thanh , Morimoto Koichi , Hosoi Yoshihiko

It has been proven that treatment of cloned embryos with a histone deacetylase inhibitor (HDACi) such as trichostatin A (TSA) following somatic-cell nuclear transfer (SCNT), significantly improves subsequent development to a blastocyst as well as full-term development (Kishigami et al. BBRC 2006). Recently, we have shown that lysine acetylation levels in both the nucleus and cytoplasm are significantly increased after oocyte activation (Matsubara et al.</...

ra0001p049 | (1) | WCRB2014

Nucleoli are formed in developing mouse embryos without nucleolus precursor bodies

Kyogoku Hirohisa , Fulka Josef , Kitajima Tomoya S , Wakayama Teruhiko , Miyano Takashi

Introduction: The large, compact oocyte nucleoli, sometimes referred to as nucleolus precursor bodies (NPBs), are essential for embryonic development in mammals. It has been convincingly documented that zygotes inherit the oocyte nucleolar material and form NPBs again in pronuclei. During early embryonic development, the compact zygote NPBs gradually transform into reticulated nucleoli of somatic cells type. Here we show that zygote NPBs are not required for embryonic and full...

ra0001p050 | (1) | WCRB2014

Fingerprints on oocyte and embryo lipid profiles caused by in vitro embryo production system and fatty acid diet supplementation in bovine

Gonzalez Serrano Andres Felipe , Ferreira Christina R , Pirro Valentina , Lucas-Hahn Andrea , Heinzmann Julia , Hadeler Klaus-Gerd , Aldag Patrick , Baulain Ulrich , Piechotta Marion , Danicke Sven , Cooks Robert Graham , Niemann Heiner

Introduction: Lipid content of oocytes and early embryos is critical for embryonic and fetal development. Indeed, the success of assisted reproductive techniques (ARTs) such as cryopreservation and in vitro embryo production is heavily affected by the lipid content of preimplantation embryos.Materials and methods: Lipid profiling of single in vitro (IVTB) and in vivo (IVVB) produced bovine embryos was performed by desorption el...

ra0001p051 | (1) | WCRB2014

Degradation of estrogen receptor α in activated blastocysts is associated with implantation in the mouse

Matsumoto Hiromichi , Saito Kyosuke , Furukawa Etsuko , Kobayashi Mitsuru , Fukui Emiko , Yoshizawa Midori

Introduction: Implantation of a blastocyst into a receptive uterus involves a series of highly coordinated cellular and molecular events directed by ovarian estrogen and progesterone. In particular, estrogen is essential for on-time uterine receptivity and blastocyst activation in mice. Although estrogen receptor α (ERα) is expressed in blastocysts, its targeted disruption leaves embryonic development and implantation unaffected. Therefore, the role of ERα in im...

ra0001p052 | (1) | WCRB2014

Production of prion gene knockout cow to prevent spontaneous bovine spongiform encephalopathy

Manabe Noboru , Onoyama Ichiro , Li Junyou , Sendai Yutaka , Aoyagi Yoshito

Introduction: In 1986, typical borine spongiform encephalopathy (BSE) was found in UK. Cattle were orally given abnormal prion protein in the brain of sheep with Scrapie. Orally administered abnormal prion protein causes variant type Creutzfeldt-Jakob disease infection in human. In 2003, spontaneous BSE was found in many countries. Spontaneous BSE is transmissible from cattle to monkey. In 2012, United States Department of Agriculture reported that spontaneous BSE cow was foun...

ra0001p053 | (1) | WCRB2014

The effect of lysophosphatidic acid during in vitro maturation of bovine cumulus–oocyte complexes: cumulus expansion, glucose uptake and expression of expansion-related genes

Boruszewska Dorota , Sinderewicz Emilia , Kowalczyk-Zieba Ilona , Woclawek-Potocka Izabela

The role of cumulus cells during in vitro maturation is essential for proper oocyte maturation and acquisition of its developmental competence. During development of ovarian follicle, cumulus cells undergo expansion, leading to extracellular matrix production (mainly hyaluronic acid), regulated by various intracellular signaling cascades. Moreover, cumulus cells metabolize glucose to pyruvate or lactate, substrates used by oocyte for its growth and maturation. We demo...

ra0001p054 | (1) | WCRB2014

Essential roles of condensins in chromosome organization during mouse early embryogenesis

Lee Jibak , Shimizu Moeko , Nishide Kenji , Hirano Tatsuya

Introduction: Multi-subunit protein complexes, called condensin I and condensin II, play a pivotal role in construction and segregation of mitotic chromosomes in many organisms. In mammals, however, previous studies examining condensins’ function are limited to siRNA-mediated knockdown experiments using some established cell lines. Hence the roles of both condensins in mammalian development have remained to be solved. In the present study, we have investigated defective p...

ra0001p055 | (1) | WCRB2014

Effect of downregulating CDX2 transcript by RNA interference on early development of bovine embryos

Sakurai Nobuyuki , Takahashi Kazuki , Hashizume Tsutomu , Sawai Ken

In Cdx2-deficient mouse embryos, Oct-4 and Nanog expression are not restricted to the inner cell mass (ICM), and trophectoderm (TE) development is impaired. These results suggest that Cdx2 regulates Oct-4 and Nanog expression in mouse embryos, and plays a key role in TE development. The objective of this study was to investigate the role of CDX2 in the early development of bovine embryos. We attempted CDX2 downregulation in bovine embryos by short interfering RNA (siRNA), and ...

ra0001p056 | (1) | WCRB2014

Fine cryopreservation method of porcine blastocysts produced by in vitro fertilization

Min Sung-Hun , Kim Jin-Woo , Lee Yong-Hee , Ahn Jae-Hyun , Do Geon-Yeop , Chae Sung-Kyu , Park Humdai , Koo Deog-Bon

Introduction: Cryopreservation has been applied successfully in many mammalian species. Nevertheless, pig embryos have shown a reduced ability to be a lower developmental competence, because of their greater susceptibility to cryoinjuries. The aim of this study was to evaluate the survival status of vitrified–warmed porcine blastocysts.Materials and methods: Forced collapse blastocoel (FBC) and non-FBC blastocysts are vitrified and concomitantly cul...

ra0001p057 | (1) | WCRB2014

In vivo embryo production in queens treated with deslorelin acetate (Suprelorin)

Louise Ackermann Camila , Ferrari Crocomo Leticia , Trevisol Eduardo , Silva Rascado Tatiana , Volpato Rodrigo , Lopes Carlize , de Freitas Guaitolini Carlos Renato , de Almeida Costa Talita , Lopes Maria Denise

Introduction: Reversibility after use of deslorelin acetate as a contraceptive for queens has been described. However no data concerning the ability of in vitro embryo production after treatement was published.Objective: This study aimed to evaluate the in vitro embryo production in queens treated with deslorelin acetate (Suprelorin).Methodology: Twenty queens and one tomcat were used. Ten queens were treated with...

ra0001p058 | (1) | WCRB2014

O-GlcNAcylation in pig embryos during preimplantation development

Shibutani Mihiro , Miyano Takashi , Miyake Masashi

Protein modification with O-linked β-N-acetylglucosamine (O-GlcNAcylation) is essential for eukaryotic cells. There are many reports concerned with O-GlcNAcylateion, but little is known about it in preimplantation development. The objective of this study was to examine the presence of O-GlcNAcylation and the role of O-GlcNAc cycling in pig preimplantation development using parthenogenetic diploids. In-vitro matured ooc...

ra0001p059 | (1) | WCRB2014

Importance of amino acids in the development of preimplantation mouse embryo

Zamfirescu Radu , Shreedharan Salini , Zada Mark , Morris Michael , Day Margot L

The composition of the external environment can have a significant impact on intracellular signaling pathways during early embryonic development. Components of the culture media, especially amino acids, are known to improve embryo development. We have shown that L-proline and L-glutamine improve development to the blastocyst stage by acting in a growth factor-like way. The aim of this study is to investigate the effect of amino acids on signaling pathways in the...

ra0001p060 | (1) | WCRB2014

Thermoprotective action of IGF1 in embryos hamster 4-cells subjected to heat stress in vitro: effect of the redistribution of E-cadherin

Cordova Alfredo Trejo , Benitez Ismael Abad , Victor Manuel Meza Villalvazo , del Carmen Navarro Maldonado Maria , Garcia Demetrio Ambriz

Introduction: IGF1 has been used as a thermoprotective during in vitro culture, however, its function is dependent on the cleavage embryos. Furthermore, the localization of E-cadherin dependent cleavage embryos. It is unknown whether there is a relationship between the thermoprotective action of the IGF1 and localization of E-cadherin.Materials and methods: Ten female hamster young were used. Hamster embryo 4-cells were obtained. Groups of embry...

ra0001p061 | (1) | WCRB2014

Effect of blastocyst artificial collapse prior to vitrification on pluripotency-specific genes expression in mouse embryos

Dashtizad Mojtaba , Shamsara Mehdi , Daliri Morteza , Zandi Ghazaleh , Fathalizadeh Parisa , Hashemi Ehsan , Hajarian Hadi

Introduction: Cryopreservation is possible for all stages of pre-implantation embryos. It has been reported that survival rate of blastocyst is comparably lower than other stages. There is a high volume of fluid in blastocoel cavity that can be a good ground for ice crystals formation, resulting in damage to the cell structure. In this study, the effects of artificial collapseand reduction of the fluid volume in blastocyst cavity before vitrification process on the survival ra...

ra0001p062 | (1) | WCRB2014

The role of histone H3 variants in the alteration of chromatin structure after fertilization

Kawamura Machika , Aoki Fugaku

After fertilization, differentiated oocytes become totipotent embryos. In this transition, a drastic change in chromatin structure is known to occur, although the mechanism underlying this change remains to be elucidated. Recent studies have revealed that one of the major factors that contribute to the change in the chromatin structure is the exchange of histones with their variants. In mammals, there are three main non-centromeric histone H3 variants: H3.1, H3.2, and H3.3. Us...

ra0001p063 | (1) | WCRB2014

Roles of the untranslated regions in the translational regulation of cyclin A2 in murine oocytes and embryos

Mori Yuuka , Aoki Fugaku

After the resumption of meiosis and fertilization, differentiated oocytes dramatically change their nature to generate totipotent zygotes. However, since transcription does not occur in oocytes and early 1-cell stage embryos, gene expression is controlled only by post-transcriptional regulation. There are huge mRNAs stored in full-grown oocytes (FGO). A part of them is not translated in FGO and MII stage oocytes, and become translated after the resumption of meiosis and fertil...

ra0001p064 | (1) | WCRB2014

The effects of PHE mixture, theophylline, and sperm concentrations on fertilization and development of bovine oocytes in vitro

Kang Sung-Sik , Yanagawa Yojiro , Nagano Masashi

Introduction: We aimed to establish an efficient in vitro fertilization (IVF) system without optimization of sperm concentration of individual bulls, which can obtain stable fertilization and development. We examined the effects of PHE mixture (20 μM D-penicillamine, 10 μM hypotaurine, and 1 μM epinephrine), theophylline (2.5 mM), and sperm concentrations (1, 2, or 5×106 spermatozoa/ml) on fertilization and development of bovine oocytes ...

ra0001p065 | (1) | WCRB2014

The effects of mesenchymal stem cell- conditioned medium on the fertilization rate, embryo development and OCT-4 gene expression of 8-cell embryo in NMRI mice

Azizi Alireza , Shahbazi Majid , Haidari Kamran

Normal growth of oocyte, embryo and also successful Implantation depend on environmental factor such as the secretion of cumulus cells and composition of follicular and tubular fluids.Therefore, addition of serum, conditioned medium (CM) to the culture medium and also co-culture with somatic cells improve mammalian embryo development.So, in the respect to the secretion of different growth factors and cytokines by mesenchymal stem c...

ra0001p066 | (1) | WCRB2014

The regulatory roles of let-7 in embryo dormancy in mice

Liu Weimin , Pang Ronald T K , Yeung William S B

Introduction: MicroRNAs interact with multiple mRNAs resulting in their degration and/or translational repression. Embryo diapause is a widespread phenomenon in which temporarily arrest occurred in embryo development. Our previous data showed that the levels of let-7 are relatively high in diapause embryos compared to reactivated embryos by E2 in mice. However, it is still not clear whether or not let-7 is involved in embryo diapause.Materials...

ra0001p067 | (1) | WCRB2014

Plasma growth hormone decline during early mammalian development is the result of expanding blood volume

Menzies Brandon R , Hetz Jennifer , Rao Alexandra , Clarke Iain , Shaw Geoff , Renfree Marilyn B

Growth hormone (GH) is necessary to grow to normal adult size. While GH plasma concentrations are highest during early development, there is no evidence that GH influences somatic growth until after birth when GH-receptors are upregulated in peripheral tissues. The gradual decline in plasma GH during fetal life in a range of mammals suggests that negative feedback to the developing pituitary occurs progressively to decrease pituitary GH output. We have tested this hypothesis u...

ra0001p068 | (1) | WCRB2014

Single-step generation of rabbits carrying a targeted allele using CRISPR/Cas9

Honda Arata , Hirose Michiko , Sankai Tadashi , Yasmin Lubna , Yuzawa Kazuaki , Honsho Kimiko , Izu Haruna , Ikawa Masahito , Ogura Atsuo

Introduction: The type II bacterial clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated protein (Cas) have been proven to be an effective gene targeting system. Genome editing of non-rodent mammalian species is a promising strategy for generation of animal models for human diseases. Mashiko et al. reported a high efficacy of direct injection of a plasmid DNA, encoding humanized Cas9 and sgRNA (single-guide RNA) in mice. Here we show successf...

ra0001p069 | (1) | WCRB2014

Supplementation of 6-diazo-5-oxo-l-norleucine improves in vitro maturation and in vitro developmental competence in pigs

Kim Ji-Su , Yang Hae-Jun , Jeong Pil-Soo , Mun Seong-Eun , Choi Seon-A , Yoon Seung-Bin , Park Young-Ho , Song Bong-Seok , Sim Bo-Woong , Kim Sun-Uk , Chang Kyu-Tae

Activators of protein kinase A (PKA) signaling pathway have been frequently used to transiently stall meiosis at early phase of in vitro maturation (IVM) for balancing between cytoplasmic and nuclear maturation of mammalian embryos. Despite of the tight association of 6-diazo-5-oxo-l-norleucine (DON), an inhibitor of hyaluronan synthesis, with PKA cascade, few evidences has been provided concerning the role of DON as an IVM supplement with meiosis-stalling activity du...

ra0001p070 | (1) | WCRB2014

Treatment of fetal bovine serum improves early development of porcine embryos by alleviating oxidative stress

Choi Seon-A , Mun Seong-Eun , Jeong Pil-Soo , Yang Hae-Jun , Yoon Seung-Bin , Park Young-Ho , Song Bong-Seok , Sim Bo-Woong , Kim Ji-Su , Kim Sun-Uk , Chang Kyu-Tae , Huh Jae-Won

Despite of the application of numerous supplements to improve in vitro culture (IVC) of mammalian cells, few studies have been conducted concerning the effect of fetal bovine serum (FBS) on early development of mammalian embryos. Thus, the current study was carried out to determine the effect of FBS on the developmental competence of porcine embryos and to establish the optimal treatment conditions. Unlike the treatment during full-term (1–6 days) or early phase ...

ra0001p071 | (1) | WCRB2014

Hollow fiber vitrification of in vitro produced bovine embryos at early developmental stages

Uchikura Ayuko , Matsunari Hitomi , Nakano Kazuaki , Matsumura Yukina , Hatae Shota , Asano Yoshinori , Nagashima Hiroshi

The hollow fiber vitrificaiton (HFV) method, we developed (Matsunari et al., JRD 58, 2012) has been shown to be very effective in the cryopreservation of highly cryosensitive embryos, such as in vitro matured-fertilized pig morulae. The objective of this study was to verify the effectiveness of the HFV method to the in vitro matured-fertilized bovine embryos in early developmental stage. Crossbred (Holstein×Japanese Black) in vitro matured-ferti...

ra0001p072 | (1) | WCRB2014

Treatment of histone deacetylase inhibitor increases in vitro developmental competence of bovine cloned embryos through suppression of endoplasmic reticulum stress

Kim Sun-Uk , Jeong Pil-Soo , Yang Hae-Jun , Mun Seong-Eun , Choi Seon-A , Yoon Seung-Bin , Park Young-Ho , Song Bong-Seok , Sim Bo-Woong , Kim Ji-Su , Chang Kyu-Tae

Although the beneficial roles of HDAC inhibitors (HDACis) in somatic cell nuclear transfer (SCNT) were known, few studies have conducted concerning the role of valproic acid (VPA) as an efficient HDACi for SCNT and its associated developmental event(s). Thus, the present study was carried out to determine the effect of VPA on developmental competence of bovine SCNT embryos and the underlying mechanism(s). We showed that the VPA treatment restored the histone acetylation level ...

ra0001p073 | (1) | WCRB2014

Early development of bovine embryos depends on the cooperative action between oxidative and endoplasmic reticulum stresses

Yoon Seung-Bin , Yang Hae-Jun , Mun Seong-Eun , Jeong Pil-Soo , Choi Seon-A , Park Young-Ho , Song Bong-Seok , Sim Bo-Woong , Kim Ji-Su , Kim Sun-Uk , Chang Kyu-Tae , Lee Sang-Rae

The coupling of reactive oxygen species (ROS) and endoplasmic reticulum (ER) stress has been explored using a variety of biological systems, but little is known regarding their role in the early development of mammalian embryos. Here, we demonstrated that early embryonic development of in vitro-produced (IVP) bovine embryos was governed by the cooperative action between ROS and ER stress. Compared to 5% O2 tension, 20% O2 significantly decreased ...

ra0001p074 | (1) | WCRB2014

Novel porcine OCT4 variants identification in blastocyst and discriminative expression analysis in adult tissues

Hwang Jae Yeon , Oh Jong-Nam , Lee Dong-Kyung , Lee Chang-Kyu

Introduction: OCT4 has been known to master gene for maintaining pluripotency in mouse inner cell mass and embryonic stem cell. However, the variant of OCT4 was false-positively detected in somatic cells and this confuses the result in stem cell and embryo development research. So in this study, we aimed to identifying novel OCT4 variants in pig and examined their expression in various adult tissues.Material and method: By comparing the sequence between ...

ra0001p075 | (1) | WCRB2014

Chromatin remodelling gene Cecr2 in murine gametogenesis, fertilization, and early embryonic development

Norton Kacie A , McDermid Heather E

Mammalian reproduction is dependent on a myriad of genes, all of which must be carefully regulated spatially and temporally to ensure successful fertilization and embryonic development. Chromatin remodellers are able to affect nuclear processes such as gene transcription, DNA replication, recombination and repair through modulation of chromatin structure. Mice with hypomorphic mutations in the chromatin remodelling gene Cecr2 have normal reproductive histology and the...

ra0001p076 | (1) | WCRB2014

Are cumulus cells additional players in calcium signalling during cattle oocyte fertilisation?

Sutton-McDowall Melanie L , Thompson Jeremy G

Upon sperm entry within the oocyte, pulsatile calcium release occurs, facilitating activation of the oocyte. While calcium signalling has been characterised in mouse oocytes, patterns within the whole cumulus oocyte complex (COC) and in larger mammals such as cow, are yet to be determined. The aim of this study was to investigate calcium changes in media, cumulus and oocytes during in vitro fertilisation in cattle oocytes.Cattle COCs were insemi...

ra0001p077 | (1) | WCRB2014

Pregnancy of bovine somatic cell nuclear transfer embryos reconstructed by using donor cells: iPS cell vs AID transfected cell*

Kim Tae Suk , Baek Sang Ki , Moon Song Yi , Jin Sang Jin , Ko Yeoung-Gyu , Kim Sung Woo , Park Hae-Geum , Seong Hwan-Hoo , Lee Joon Hee

SCNT technology provides potential applications for biomedical and agriculture. However, the efficiency of SCNT is still low. Incomplete epigenetic reprogramming of transferred somatic cell is believed to be one of main causes of developmental problems. Hanwoo is considered as only bovine species with mainly reddish color but quite a few of mini-, white-colored, black-colored, striped cows. The objective of this study was to produce cloned offspring from endangered mini-Hanwoo...

ra0001p078 | (1) | WCRB2014

The effect of cRNA concentration of artificial nuclease microinjected cytoplasmically to pronuclear porcine embryos on survival and development in vitro

Kamoshita Maki , Kato Tsubasa , Sagara Eri , Hisamatsu S , Sakaue M , Sakuma Tetsushi , Yamamoto Takashi , Ito Junya , Kashiwazaki Naomi

Although GH receptor (GHR) gene deletion causes a hereditary dwarfism and also shows a longer lifespan, the fact was shown only in mice. Because it was difficult to generate knockout (KO) animals other than mice. Recently, the generation of KO animals became possible without difficulty due to advances in genome editing technologies such as transcription activator-like effector nuclease (TALEN). In the present study, we examined the effect of cRNA concentration microinjected cy...

ra0001p079 | (1) | WCRB2014

Abstract withdrawn....

ra0001p080 | (1) | WCRB2014

Isolation and characterization of porcine mesenchymal stem cell as a donor for efficient nuclear transfer

Lee Jun Sung , Jung Sung Han , Park Young Bin , Kim Sang Eun , Lee Hoon Taek

Introduction: Nuclear transfer (NT) has used for generating cloned animals or genetically modified animals. However, the efficiency has remained low, because of epigenetic errors that occur during donor cell reprogramming. Mesenchymal stem cells (MSCs) were known as undifferentiated state cell compared to somatic cells. Thus MSCs can reduce the chance of error that can be occurred during reprogramming process and can be easily isolated from adult, while embryonic stem cells ar...

ra0001p081 | (1) | WCRB2014

Establishment of efficient system in the DNA microinjection into porcine in vitro embryos

Malaweera Don Buddika Oshadi , Ramachandra Sisitha , Kim Ghangyong , Kim Seokjoong , Cho Jongki

To investigate the efficient laboratory techniques for the injection of DNA, pEGFP-N1 commercial plasmid were microinjected into porcine parthenogenetic and IVF embryos to explore the injection time, volume and concentration, for the efficient blastocyst production. In experiment 1, to investigate injection time, compared four different time durations (2, 4, 6 and 8 h) after post activation and 6 h of co-incubation with sperms. There were no significant difference (P&...

ra0001p082 | (1) | WCRB2014

The effect of development in vitro

Obata Hitomi , Kamoshita Maki , Kato Tsubasa , Ito Junya , Kashiwazaki Naomi

Although mammalian embryo culture is important for research in early development as well as reproduction engineering, rat embryo culture is still infant other than that of mice. The aim of the present study was to improve rat embryo culture in vitro, we compared modified rat-one cell embryo cuture medium (R1ECM) with potassium simplex optimized medium with amino acids (KSOMaa) and that effect of addition of EDTA (0.013 mM) and glutamine (1 mM) to mR1ECM on develop to ...

ra0001p083 | (1) | WCRB2014

Comparison of survivin gene expression between porcine SCNT and iSCNT-derived normal and arrested embryos

Choi Kyu Hee , Kim Young Kyu , Choi Seong Deok , Shin Sang Tae , Lee Hoon Taek

Introduction: Interspecies somatic cell nuclear transfer (iSCNT) is an useful tool to produce cloned embryos using livestock oocytes instead of the animal with peculiar reproductive system to produce endangered animal and rescue incurable disease. There are many researches about iSCNT such as porcine-bovine, mouse-porcine, mouse-bovine and so on, in contrast, canine-porcine SCNT especially have been almost no reports.Materials and methods: Porcine fetal ...

ra0001p084 | (1) | WCRB2014

Involvement of poly(ADP-ribosyl)ation and autophagy in porcine pre-implantation development

Lee Hye Ran , Hwang Jeongho , Park Hyo Chang , Han Yong Man , Lee Hoon Taek

Introduction: The poly(ADP-ribosyl)ation (PARylation) has involvement with pro-survival autophagy. However, there has been no report about PARylation and autophagy during pre-implantation development. Here, we investigated the mechanism between PARylation and pro-survival autophagy in pre-implantation development.Materials and methods: IVF embryos were cultured in the medium containing 3-aminobenzamide (3ABA), parp inhibitor. Autophagy and PARylation wer...

ra0001p085 | (1) | WCRB2014

Transporters for L-proline in the pre-implantation mouse embryo

Day Margot L , Zada M , Bailey Charles , Treleaven Tamara , Ozsoy Sukran , Rasko John EJ , Morris Michael B

The inclusion of amino acids in pre-implantation embryo culture media is known to improve the rate of development and embryo viability. Our studies have shown that embryos cultured in L-proline from the zygote to the blastocyst stage develop better than embryos cultured in the absence of amino acids. This study aimed to identify the pre-implantation stage(s) at which L-proline transport is required for development to be improved and then to charac...

ra0001p086 | (1) | WCRB2014

Treatment of antioxidant (acteoside) in fetal fibroblats improves the efficiency of canine cell cloning

Lee Ji Hye , Kim Keun Jung , Kim Eun Young , Xia Li Xiao , Park Kang Sun , Han Kil Woo , Lee Jin Hee , Lee Bo Myeong , Yu Jeong , Zhuang Li Li , Kim Dong Hee , Lee Kyung Bon , Kim Dong-Hoon , Kim Min Kyu

This study was designed to investigate whether acteoside (antioxidant) improves canine SCNT efficiency. Cell cycle of canine fetal fibroblasts was synchronized by culturing to contact inhibition, serum starvation and treating with acteoside. Cell cycle synchronization, apoptosis and reactive oxygen species (ROS) detection were analyzed using flow cytometry. The fibroblasts, prepared by confluent-cell culture or treating with 30 μM acteoside for 48 h, were reconstructed in...

ra0001p087 | (1) | WCRB2014

Semi-quantification and elucidation of bovine embryo biomarkers by mass spectrometry imaging

Goncalves Roseli F , Ferreira Monica S , de Oliveira Diogo N , Canevarolo Rafael R , Achilles Marcos A , Killian Gary J , Bols Peter E , Visintin Jose A , Catharino Rodrigo R

In the field of ‘single cell analysis’, many classical strategies like immunofluorescence and electron microscopy are the fundamental choices. However, these methodologies are time-consuming and do not permit direct identification of specific molecular classes such as lipids. A novel mass spectrometry-based analytical approach has been applied to bovine oocytes and embryo. This new metabolomics-based application uses mass spectrometry imaging (MSI), efficient data pr...

ra0001p088 | (1) | WCRB2014

Artificial activation enhances pre-implantation embryo development after intracytoplasmic injected with evaporative-dried sperm

Li Xiao Xia , Lee Ji Hye , Kim Keun Jung , Kim Eun Young , Park Kang Sun , Han Kil Woo , Lee Jin Hee , Lee Bo Myeong , Kim Dong Hee , Yu Jeong , Zhuang Li Li , Kim Min Kyu

Evaporative-drying is one alternative to cryopreservation. It has the advantages that during sample preparation it does not require liquid nitrogen and a freeze-dryer machine; however, the results have not been as successful as freeze-drying. The objectives of this study were to determine whether artificial activation (electrical and chemical activation) could improve the development of porcine embryos in vitro after intracytoplasmic injected with evaporative-dried sp...

ra0001p089 | (1) | WCRB2014

Construction of porcine FoxN1 knockout system by RNA-guided endonucleases

Park Jae-Kyung , Jang Hoon , Kim Donghwan , Sun Wu Sheng , Kim Hyoung-Joo , Yeo Jaehun , Hwang Seongsoo , Lee Jeong-Woong

Introduction: Forkhead box protein N1 (FoxN1) regulates development, differentiation, and function of thymus epithelial cells (TECs), both in the prenatal and postnatal thymus. In mice, knockout of the FoxN1 results in two well-known defects as hairlessness and athymia. However, knockout of FoxN1 in other species has not been discovered yet. RNA-guided endonucleases (RGENs), derived from the prokaryotic type II CRISPR-Cas system, enable targeted specific genes. We successfully...

ra0001p090 | (1) | WCRB2014

Expression pattern and function of claudins during tight junction maturation in the mouse pre-implantation embryo

Sheth Bhavwanti , Cox Andrew L , Greenslade Karen J , Alexopoulos Orpheas , Fleming Tom P

Introduction: The pre-implantation embryo has two distinct cell lineages at the blastocyst stage. An outer epithelial monolayer called trophectoderm and the non-polar inner cell mass. Tight junctions (TJ) form continuous intercellular contacts between neighbouring trophectoderm cells and are crucial in establishing a blastocoele cavity. TJs are composed of several transmembrane proteins including Claudins, a large family of proteins that play a central role in the formation of...

ra0001p091 | (1) | WCRB2014

Human granulocyte–colony stimulating factor enhances viability of porcine embryos in defined oocyte maturation medium

Cai Lian , Hyun Sang-Hwan

Granulocyte colony-stimulating factor (G-CSF) enhances the proliferation, differentiation and survival of cells. In addition, G-CSF is a non-invasive biomarker of human oocyte developmental competence for embryo implantation. In human follicle G-CSF concentration increased during the ovulatory. At present experiment, the cumulus oocyte complexes (COCs) were aspirated from superficial follicles (1–3 and 3–6 mm). COCs from small and medium follicle were matured in prot...

ra0001p092 | (1) | WCRB2014

Chromatin remodelling and histones mRNA accumulation in bovine germinal vesicle oocyte

Labrecque Remi , Lodde Valentina , Dieci Cecilia , Tessaro Irene , Luciano Alberto M , Sirard Marc-Andre

In several mammalian species, a major remodelling of the germinal vesicle chromatin is known to occur towards the end of the oocyte growth. Various chromatin configurations have been identified. However, the mechanisms involved in this remodelling process are yet not completely understood. In the bovine species, four distinct and progressive states of chromatin compaction have been characterized and are linked to a gradual acquisition of the developmental potential. Germinal v...

ra0001p093 | (1) | WCRB2014

Effect of ganglioside GT1b treatment during porcine in vitro maturation on embryonic development and mRNA expression pattern in cumulus cell

Hwang Seon-Ung , Hyun Sang-Hwan

Ganglioside is an acidic glycosphingolipid with sialic acid residues. The b-series ganglioside GT1b was reported that suppressing damage of mtDNA by reactive oxygen species (ROS) in mouse brain. The purpose of this study is to investigate the effect of exogenous addition of GT1b on in vitro maturation (IVM) of porcine oocytes and to confirm the related bradykinin 2 receptors (B2R). GT1b were treated on IVM that concentration was 0 (control), 5, 10 and 20 nM. After IVM...

ra0001p094 | (1) | WCRB2014

The antioxidative effect of carboxyethylgermaniumsesquioxide (Ge-132) on in vitro maturation of porcine oocytes and subsequent embryonic development after in vitro fertilization and parthenogenetic activation

Kim Eunhye , Hyun Sang-Hwan

The carboxyethylgermaniumsesquioxide (Ge-132) is an organogermanium compound known to have anti-oxidative effect. In this study, we examined the effect of Ge-132 on in vitro maturation (IVM) of porcine oocytes analyzing intracellular glutathione (GSH) and reactive oxygen species (ROS) levels, mRNA expression levels and subsequent embryonic development after in vitro fertilization (IVF) and parthenogenetic activation (PA). After 40 h of IVM, intracellular GSH ...

ra0001p095 | (1) | WCRB2014

The effect of vitrification on subsequent mouse embryo development and postnatal health

Khalif Raja Ili Airina Raja , Smyth Neil , Fleming Tom , Velazquez Miguel Abraham , Sheth Bhav , Smith Stephanie

Short-term effects of cryopreservation on embryo development, survival and after embryo transfer (ET), resulting pregnancy rate have been documented. However, long-term effects on offspring remain relatively unexplored.Materials and methods: Using a mouse model, we investigated the effect of vitrification on blastocyst trophectoderm (TE) and inner cell mass (ICM) cell numbers by differential staining and postnatal growth and cardiovascular and metabolic ...

ra0001p096 | (1) | WCRB2014

Insulin and branched-chain amino acid depletion during mouse in vitro preimplantation development alters postnatal growth and cardiovascular physiology

Velazquez Miguel Abraham , Sheth Bhav , Marfy-Smith Stephanie , Eckert Judith , Fleming Tom

Introduction: Maternal depletion of insulin in serum and branched-chain amino acids (BCAA) in uterine luminal fluid occurs in response to dietary protein undernutrition during the periconceptional period which leads to adverse offspring growth and cardiovascular phenotype (e.g. high blood pressure). This study aimed to determine whether insulin and/or BCAA depletion might be inductive in vivo factors for postnatal phenotype using an in vitro embryo culture mo...

ra0001p097 | (1) | WCRB2014

Supplementation with sunflower seed increases conception rates in recipient beef heifers

Membrive Claudia Maria Bertan , Cordeiro Mariangela Bueno , de Souza Johnny Maciel , Filho Manoel Francisco Sa , Pugliesi Guilherme , de Paula Nogueira Guilherme , Santin Tiago , Binelli Mario

In a previous study (Peres et al., 2008 Acta Scientiae Veterinariae 36 639), sunflower supplementation during 22 days improved conception rates by 20.4% in Timed AI programs in beef cows. We aimed to evaluate the effects of sunflower seed supplementation on circulating cholesterol (CHOL) and progesterone (P4) concentrations, and the conception rate of recipient beef heifers after timed embryo transfer (TET). Heifers were synchroniz...

ra0001p098 | (1) | WCRB2014

The role of vitamin C in in vitro early embryonic development: verification using aldehyde reductase gene knockout mice

Hoshino Yuki , Watanabe Ren , Takahashi Motoko , Miyata Satoshi , Yoshida Yasukazu , Fujii Junichi , Kimura Naoko

Introduction: Supplementation of appropriate amount of vitamin C (VC) improves oocyte fertilization and subsequent embryo development while high concentrations of VC rather reduce the developmental potency. To clarify the role of VC, oocytes were collected from aldehyde reductase (AKR1A) knockout mice that are unable to synthesize VC and subjected to analyses from view point of developmental potency and gene expression involved in VC metabolism.Materials...

ra0001p099 | (1) | WCRB2014

Selection of reference genes in mouse preimplantation embryos of different ploidies at various developmental stages

Gu Yanli , Shen Xinghui , Zhou Dongjie , Wang Zhendong , Lei Lei

Real-time RT quantitative PCR (qPCR) has become the most frequently used system for studies of gene expression. Many studies have provided reliable evidences that the transcription levels of reference genes are not constant at different developmental stages and in different experimental conditions. However, suitable reference genes which are stably expressed in polyploid preimplantation embryos of different developmental stages have not yet been identified. In the present stud...

ra0001p100 | (1) | WCRB2014

Competence of porcine first polar body for normal development

Jin Dong Il , Lin Tao , Diao Yun Fei , Won Kang Jung , Kim Dong Kyo

To determine whether chromosomes in porcine first polar body (PB1) can complete the second meiotic division and participate in subsequent normal embryo development, we examined developed competence of chromosome in PB1 that had been injected into enucleated MII stage oocytes by nuclear transfer method (chromosomes replacement group, CR group). After parthenogenetic activation (PA) and in vitro fertilization (IVF), the cleavage rate of reconstructed oocytes in IVF grou...

ra0001p101 | (1) | WCRB2014

Association between birth weight and age at menarche: a meta-analysis

Yermachenko Anna , He Hao , Dvornyk Volodymyr

Low weight at birth may affect the onset of sexual maturation. Early puberty timing in those being small for gestational age might be explained by fetal reprogramming that helps to adapt for the poor-resource postnatal life and be ready earlier for reproduction. Biological plausibility for this association prompted for many epidemiological studies aimed to test a relationship between low birth weight and age at menarche (AAM) as a reliable indicator of puberty timing.<p cl...

ra0001p102 | (1) | WCRB2014

Effect of downregulating GLIS1 transcript on early development and gene expression of bovine embryos

Takahashi Kazuki , Sakurai Nobuyuki , Hashizume Tsutomu , Sawai Ken

GLIS1 markedly enhances the generation of iPS cells from both mouse and human fibroblasts. In mouse, GLIS1 is enriched in unfertilized oocytes and 1-cell stage embryos. Therefore, it is possible that GLIS1 plays a critical role in early development through epigenetic regulation. We investigated the role of GLIS1 during the early development of bovine embryos using siRNAs targeted for GLIS1. GLIS1 transcript levels in unfertilized oocytes and early embryos at ...

ra0001p103 | (1) | WCRB2014

Genome-wide reprogramming by DNA demethylation during mouse oocyte growth and early development

Sakashita Akihiko , Iseki Yosuke , Nakajima Mei , Wakai Takuya , Kobayashi Hisato , Kono Tomohiro

Introduction: Genome-wide demethylation reprograms the genome after fertilization and re-establishes totipotency: 5-methylcytosine (5 mC) in the paternal pronucleus is rapidly converted to 5-hydroxymethylcytosine (5 hmC) by the dioxygenase 10–11 translocation (TET) 3, while 5 mC in the maternal pronucleus is protected by the binding of developmental pluripotency-associated (DPPA) 3 to histone H3 dimethyl Lys9 (H3K9me2). This process is essential for normal development, be...

ra0001p104 | (1) | WCRB2014

Expression profile of imprinted genes and DNA methyl transferases in uniparental sheep placenta

Zacchini Federica , Toschi Paola , Czernik Marta , Loi Pasqualino , Ptak Grazyna Ewa

Epigenetic programming has a crucial role in mammalian development. It is well described that disarrangement of epigenetic programming is associated with impaired embryonic development, compromised pregnancy (ie. abortion or preeclampsia) and several imprinting disorders (i.e Prader Willi and Beckwith-Wiedemann syndromes). A useful tool to better understand the etiology of compromised pregnancy is represented by uniparental embryos, characterized by only maternal (parthenogeno...

ra0001p105 | (1) | WCRB2014

Genome editing technology using ZFN and TALEN is effective on cultured porcine ST cells knockout pig

Kato Tsubasa , Sakaue M , Hisamatsu S , Kamoshita M , Obata H , Sakuma Tetsushi , Yamamoto Takashi

In recent years, advances in molecular biology and reproductive engineering make generation of knockout (KO) animals possible. KO animals are useful and powerful tools for agriculture as well as biomedical research. Zinc finger nuclease (ZFN) and transcription activator-like effector nuclease (TALEN) genome editing technology enables site directed engineering of the genome. The aim of the present study was to ZFN and TALEN system could work on editing genome of cultured porcin...

ra0001p106 | (1) | WCRB2014

Genome-wide DNA methylation analysis of bovine clones derived from the same donor cells

Kaneda Masahiro , Watanabe Shinya , Hirako Makoto , Akagi Satoshi , Kiefer Helene , Jouneau Luc , Martin-Magniette Marie-Laure , Balzergue Sandrine , Nagai Takashi

Introduction: What is nature versus nurture? To what extent do genetic inheritance and non-genetic factors contribute to one’s character? This is one of the oldest issues in psychology. To answer this historic question, cloned animals are good models because they have identical genetic information to the donors. Previous studies indicated that cloned animals showed different coat color patterns, noseprints and characters compared to the donors. However, no molecular work ...

ra0001p107 | (1) | WCRB2014

DNA methylation analysis in sperm from boars exhibiting normal and altered spermograms

Congras Annabelle , Yerle-Bouissou Martine , Vignoles Florence , Lhuillier Emeline , Foissac Sylvain , Pinton Alain , Ferchaud Stephane , Bouchez Olivier , Riquet Juliette , Acloque Herve

In the last decade, more and more studies have reported aberrant pattern of methylation in sperm DNA of patients with an altered spermogram. It is also admitted that epigenetic reprogramming during germ cell development is a key mechanism for the production of functional gametes and the proper development of the embryo. As poor semen quality is also a key issue for farm animals’ productivity, we studied the methylation profile of pig sperm DNA of fertile and poor-quality ...

ra0001p108 | (1) | WCRB2014

Effect of the way in which ewes were handled during mid-gestation on testicular and ovarian development in their lambs

de Jonge Suzanne , Hogg Charis O. , Baxter Emma , Hall Sarah , Dwyer Cathy M. , Ashworth Cheryl Joy

Introduction: This study determined whether the way in which pregnant ewes were handled affected gonad development in their offspring.Methods: Twin-bearing Scottish Blackface × Border Leicester ewes were either handled minimally (control, C, n=20), positively (predictable timings, soft vocal tones etc. P, n=20) or negatively (random timings, shouting and whistling etc. N, n=20) between day 65 and 100 of pregnancy. Followin...

ra0001p109 | (1) | WCRB2014

Postovulatory ageing influences methylation marks and development in bovine oocytes and early embryos

Heinzmann Julia , Mattern Felix , Aldag Patrick , Lucas-Hahn Andrea , Wrenzycki Christine , Haaf Thomas , Niemann Heiner

Introduction: Oocytes that are not fertilized in a specific time-frame after ovulation undergo a time-dependent process of degradation and loss of developmental potential that can be referred to as postovulatory ageing. This process occurs in vivo and under specific in vitro settings. Here, we studied epigenetic changes related to postovulatory ageing.Materials and methods: Bovine oocytes were collected from slaughterhouse ovaries and m...

ra0001p110 | (1) | WCRB2014

Epigenetic regulation of Gata-6 transcription factor expression in mouse primitive endoderm extra-embryonic lineage induced by maternal protein restricted diet

Sun Congshan , Sheth Bhav , Smyth Neil , Cox Andrew L. , Denisenko Oleg , Fleming Tom

Introduction: Maternal low protein diet exclusively during preimplantation mouse development (Emb-LPD) induces compensatory changes in extra-embryonic lineages including primitive endoderm (PE) to enhance nutrient delivery during later gestation, for example, stimulation of endocytosis. Whilst such responses protect fetal growth, adult offspring exhibit increased cardiometabolic disease risk. Here, we investigate epigenetic mechanisms underlying maternal dietary programming, u...

ra0001p111 | (1) | WCRB2014

Lactation litter size differentially affects satiety hormone concentrations and gut adaptations in Wistar rat dams

Johnson Michelle L , Saffrey M. Jill , Taylor Vicky J.

Introduction: Peptide-YY (PYY) is a satiety hormone secreted by the colon. We have previously shown that PYY significantly increases in early lactation in rats, despite hyperphagia. Increased PYY may be associated with significantly increased gut growth by late lactation in dams with average-sized litters. Our aim was to determine whether feeding different litter sizes influences postpartum changes occurring during the maternal adaptation to lactation.Ma...

ra0001p112 | (1) | WCRB2014

TAF4b promotes oocyte survival and proper ovarian chromatin state in the mouse

Grive Kathryn Jennifer , Ribiero Jennifer R. , Seymour Kimberly A. , Freiman Richard N.

Introduction: The condition of primary ovarian insufficiency (POI) affects 1% of women worldwide under the age of 40, and is associated with premature ovarian follicle depletion. TAF4b is a gonadal-enriched TFIID subunit that promotes healthy ovarian aging and fertility. Female TAF4b-deficient mice experience POI hallmarks including infertility, poor oocyte quality, and dramatic gene expression changes. Understanding TAF4b’s role in promoting healthy ovarian aging is crit...

ra0001p113 | (1) | WCRB2014

Combinations of 17β-estradiol and androgen support bovine oocyte growth in vitro by maintaining physical connections with granulosa cells

Makita Miho , Miyano Takashi

Introduction: Recent studies have revealed the contribution of steroid hormones to in vitro growth of oocytes in different species. However, exactly how these steroid hormones support oocyte growth is not well understood. This study was conducted to examine the effect of various steroid hormones on in vitro growth of bovine oocytes and to determine the contribution of steroid hormones to oocyte growth by assessing the connections between oocytes and granulosa...

ra0001p114 | (1) | WCRB2014

Anti-Mullerian Hormone (AMH) regulates the growth and maturation of primate antral follicles during the menstrual cycle

Xu Jing , Lawson Maralee S. , Bishop Cecily V.

Introduction: Studies in primates indicated that AMH promotes preantral follicle growth, but inhibits antral follicle estradiol (E2) production in vitro. Thus, experiments were designed to investigate the role of AMH on primate follicular development in vivo during the spontaneous menstrual cycle.Materials and methods: Hemi-ovariectomized, adult rhesus macaques (n=5) first received vehicle (control) treatment by intraovarian infusion from cycle ...

ra0001p115 | (1) | WCRB2014

Critical role of decoy receptor 3 in regulation of granulosa cell apoptosis in pig ovarian follicle

Fukumoto Yoshinosuke , Sugimoto Miki , Onoyama Ichiro , Wongpanit Kannika , Manabe Noboru

Introduction: During follicular development in mammalian ovaries, the majority of follicles undergo atresia. One of the characteristics of this process is apoptotic cell death in granulosa cells. Death ligands and receptors, including Fas ligand (FasL) and Fas, have been detected in follicles and also demonstrated to be capable of inducing apoptosis in follicular cells. Decoy receptor 3 (DcR3) competes with Fas to bind FasL but lacks intracellular death domains, thus inhibitin...

ra0001p116 | (1) | WCRB2014

Expression and function of lysophosphatidic acid in theca cells of the bovine ovarian follicle

Sinderewicz Emilia , Boruszewska Dorota , Kowalczyk-Zieba Ilona , Staszkiewicz Joanna , Grycmacher Katarzyna , Woclawek-Potocka Izabela

In cow, lysophosphatidic acid (LPA) stimulated synthesis and secretion of luteotropic factors in ovary, which affected growth and development of ovarian follicles. Theca cells, among other components of ovarian follicle, play an essential role in ovulation, which depends on intrafollicular prostaglandin synthesis. In bovine reproductive tract, modulation of prostaglandin synthesis occurs under influence of LPA. Thus, the aim of this study was to determine possible correlation ...

ra0001p117 | (1) | WCRB2014

Expression and function of lysophosphatidic acid in granulosa cells of the bovine ovarian follicle

Woclawek-Potocka Izabela , Sinderewicz Emilia , Boruszewska Dorota , Kowalczyk-Zieba Ilona , Staszkiewicz Joanna , Grycmacher Katarzyna

Mural granulosa cells participate in development and differentiation of ovarian follicle. In bovine reproductive system, LPA is involved in autocrine and paracrine signaling, through LPA receptors (LPAR)s. We demonstrated before that LPA modulated prostaglandin (PG) synthesis in uterus and ovary and stimulated estradiol (E2) synthesis in granulosa cells in bovine follicle. As the ovarian follicle development and differentiation leading to oocyte ovulation depend on ...

ra0001p118 | (1) | WCRB2014

Evaluation of oocyte quality and ovarian function in aged female cystine-glutamate transporter gene-deficient mice

Watanabe Ren , Fujii Junichi , Sato Hideyo , Kimura Naoko

Introduction: Cystine-glutamate transporter (xCT) regulates cysteine levels by limiting uptake of cystine and consequently determines glutathione (GSH) contents in the cells. This study was performed to clarify effects of xCT deficiency (KO) on oocyte quality and ovarian function during maternal aging.Materials and methods: Ovaries and plasma were collected from young (2 months) and aged (12 months) C57BL/6 xCT-KO and WT mice at 44 h after PMSG treatment...

ra0001p119 | (1) | WCRB2014

Ovarian and hormonal responses of female goats to feed restriction and intermittent administration of neurokinin B analog

Endo Natsumi , Tanaka Tomomi

The aim of this study was to determine the effect of feed restriction on follicular dynamics and hormone secretion patterns in goats, and to examine the effect of neurokinin B analog (senktide) under this experimental setting. Goats were administered 2 mg of dinoprost during the luteal phase, and CIDR was inserted for 10 days. Feed was provided at the level of 50% (feed-restricted; n=9) or 100% maintenance (control; n=4) for 15 days, beginning on the day of C...

ra0001p120 | (1) | WCRB2014

Differential changes in LH secretion after administration of investigational metastin/kisspeptin analog TAK-683 to goats

Tamesaki Chie , Endo Natsumi , Ohkura Satoshi , Wakabayashi Yoshihiro , Matsui Hisanori , Tanaka Akira , Watanabe Tatsuya , Okamura Hiroaki , Tanaka Tomomi

The aim of the present study was to evaluate ovarian and hormonal responses to the administration of metastin/kisspeptin analog, TAK-683, under the endocrine environments of luteal and follicular phases in goats. At 7–10 days after the confirmation of ovulation by ultrasonography, six goats received a prostaglandin F2α (PGF2α) injection (Day 0) followed by 10 days of CIDR-G® treatment. PGF2α was injected agai...

ra0001p121 | (1) | WCRB2014

Endocrine responses and ovarian dynamics in goats treated with low dose of investigational metastin/kisspeptin analog TAK-683 in follicular phase

Gomi Nahoko , Endo Natsumi , Ohkura Satoshi , Wakabayashi Yoshihiro , Matsui Hisanori , Matsumoto Hirokazu , Ishikawa Kaori , Tanaka Akira , Watanabe Tatsuya , Okamura Hiroaki , Tanaka Tomomi

The aim of the present study was to evaluate ovarian and hormonal responses to follicular phase administration of a metastin/kisspeptin analog, TAK-683, with a low dose, determined as the minimally effective dose for LH stimulation in ovariectomized goats. At 7–10 days after ovulation, goats received a prostaglandin F2α (PGF2α) injection (day 0) followed by 10 days of CIDR-G® treatment. A PGF2α was injected a...

ra0001p122 | (1) | WCRB2014

TLR4-mediated signaling pathway is modulated by melatonin through MyD88-dependent pathway in ovarian carcinoma of ethanol-consuming rats

Chuffa Luiz Gustavo , Fioruci-Fontanelli Beatriz , Mendes Leonardo , Pinheiro Patricia Fernanda , Martinez Marcelo , Martinez FranciscoEduardo

Introduction: Toll-like receptors (TLRs) are active molecules expressed on the surface of ovarian cancer (OC) cells, but the consequences of TLR2/TLR4 signaling pathways in these cells remain unclear. Because melatonin (mel) act as an immunomodulatory hormone and has been reported to modulate TLRs in some aggressive tumor cells, we investigated the effective role of long-term mel therapy on TLR2- and TLR4-mediated molecule myeloid differentiation factor 88 (MyD88)-dependent si...

ra0001p123 | (1) | WCRB2014

Relative transcript abundance in porcine cumulus cells collected from different size follicles

Nicolas Carla Moros , Rico Maria Jose Izquierdo , Li Y , Romar Rakel , Funahashi H

Introduction: Bi-directional communication between the oocyte and cumulus cells (CCs) is essential for the production of competent oocytes. Previous studies analyzed the relative transcript abundance in oocytes from small follicles (SF) and medium follicles (MF) to determinate the likely use of oocytes from SF in artificial reproductive techniques. The aim of this study was to examine the relative transcript abundance of CCs from cumulus–oocyte complexes (COCs) from SF an...

ra0001p124 | (1) | WCRB2014

Real-time observation of hormone-secretion using CFP, YFP-fused gonadotropin expression vector

Takanashi Haruka , Nishimura Naoto , Yoshida Saishu , Higuchi Masashi , Kawai Kohei , Ueharu Hiroki , Kato Takako , Kato Yukio

Introduction: FSH and LH are glycoprotein hormones and are synthesized in the anterior lobe of the pituitary gland and plays critical roles for maintenance of vital functions. FSH and LH are secreted to pituitary capillary vessel and exert their effects on reproductive organs of both sexes to stimulate steroidogenesis and gametogenesis. Secretion of FSH and LH is controlled mainly by GnRH released from hypothalamus. Many investigations on secretion of FSH and LH were performed...

ra0001p125 | (1) | WCRB2014

BMP15 plus GDF9 and cAMP modulator promote EGF receptor signaling to increase oocyte developmental competence in porcine cumulus–oocyte complexes from small follicles

Sugimura Satoshi , Ritter Lesley J , Rose Ryan D , Thompson Jeremy G , Mottershead David G , Gilchrist Robert B

Introduction: Oocytes acquire developmental competence with progressive folliculogenesis. Cumulus–oocyte complexes (COCs) from small antral follicles are commonly used in IVM but have inherent low competence and are poorly responsive to amphiregulin (AREG) which normally mediates oocyte maturation. Here we examined effects of bone morphogenetic protein 15 (BMP15), growth differentiation factor 9 (GDF9), and dbcAMP on the maturation and subsequent developmental competence ...

ra0001p126 | (1) | WCRB2014

TGFβ regulation of the human fetal ovarian matrix

Bayne Rosemary A L , Roussi Kalliopi , Childs Andrew , Rodgers Ray , Anderson Richard

Introduction: Human germ cells develop in synchronised nests in the fetal ovary, interspersed with somatic cells prior to reorganisation to form primordial follicles. We have isolated several somatic cell lines from disaggregated human fetal ovaries as a model for understanding their contribution to the germ cell niche. Fetal dysregulation of TGFβ signalling has been linked with fibrosis in polycystic ovary syndrome (PCOS): effects of TGFβ on extracellular matrix (EC...

ra0001p127 | (1) | WCRB2014

Abstract withdrawn....

ra0001p128 | (1) | WCRB2014

Effects of the mycotoxin, deoxynivalenol, on bovine granulosa cells

Guerrero-Netro Hilda Morayma , Chorfi Younes , Price Christopher A

Introduction: Deoxynivalenol (DON) is a major mycotoxin found in animal feed, one of its major actions is to activate the ribotoxic stress response (RSR) in different cells types. DON has been shown to inhibit progesterone secretion from porcine granulosa cells, but its effect in the cow is unknown. The objective of this study was to determine the effects of DON on bovine granulosa cells in vitro.Materials and methods: Bovine granulosa cells fro...

ra0001p129 | (1) | WCRB2014

Transcriptome in small antral follicles of monkeys on a western-style diet with/without testosterone

Xu Fuhua , Xu Jing , Bishop Cecily V , Cameron Judy L , Stouffer Richard L

Introduction: Recent studies indicate that dietary habits and hyperandrogenemia influence ovarian function in women. However, data are limited regarding the effects of western-style diet (WSD) and androgen (e.g. T) on follicle development, especially during the early follicular stage, in primates. The current study investigated the impact of WSD alone and with chronic T exposure on gene expression in small antral follicles (SAFs) of macaques.Materials an...

ra0001p130 | (1) | WCRB2014

The effects of estradiol and ECG combination on ZP3 gene in mouse oocyte

Mohammadzadeh Masoomeh , Shahbazi Majid , Haidari Kamran

Introduction: The ovarian stimulators have a crucial role in steriodogenesis, follicular development, oocyte maturation and embryo maintenance by activating some genes in ovarian follicles. As, one of the most important cellular and molecular mechanisms involved in successful fertilization is the expression of sperm receptor binding protein or ZP3 in zona Pellucida, therefore our attempts have been made to delineate the effects of combined Estradiol and eCG on ZP<su...

ra0001p131 | (1) | WCRB2014

Inhibin α induced porcine granulosa cells apoptosis through mitochondrial apoptotic pathway

Li Wan-Hong , Chen Shu-Xiong , Chen Lu , Li Chunjin , Zhou Xu

Inhibin is a heterodimeric gonadal glycoprotein hormone belonging to the TGFβ superfamily that named for its ability to suppress FSH synthesis and secretion from anterior pituitary via a negative feedback. It can regulate proliferation, differentiation, and steroidogenesis of granulosa cells (GCs) through paracrine and autocrine manners. The objective of this study was to investigate the interaction between inhibin α subunit (INHA) and apoptosis of porcine ovarian GC...

ra0001p132 | (1) | WCRB2014

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ra0001p133 | (1) | WCRB2014

The effect of bisphenol A on Notch signaling pathway in the follicular development of neonatal rat ovary

Altunbas Korhan , Celik Sefa , Yagci Artay , Akkaya Ozlem Ozden

Introduction: Neonatal exposure to bisphenol A (BPA) can impair reproductive physiology, but the specific mechanisms by which this occurs remain largely unknown. We aimed to study whether mRNA levels of Notch receptor and ligands are altered following BPA treatment during postnatal days.Materials and methods: Neonatal rats were divided into two main groups as groups I and II according to treatment time intervals consisting of postnatal day (P)0–P3 a...

ra0001p134 | (1) | WCRB2014

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ra0001p135 | (1) | WCRB2014

Interaction between galectin-1 and glycoconjugates is involved in hCG-mediated gene expression in cultured human luteinized granulosa cells

Nio-Kobayashi Junko , Iwanaga Toshihiko , Duncan W Colin

Introduction: The corpus luteum (CL) is a temporal endocrine organ secreting large amount of progesterone essential for the establishment and maintenance of pregnancy. The CL undergoes luteolysis in a non-fertile cycle, while the CL is rescued from luteolysis by human chorionic gonadotrophin (hCG) secreted from the conceptus during pregnancy. We recently revealed that a β-galactoside-binding lectin, galectin-1, plays important roles in the regulation of luteal rescue in t...

ra0001p136 | (1) | WCRB2014

Expression of steriod hormone receptor in the ovary and oviduct during the canine estrus cycle

Yang Byoung-Chul , Yoo Jae Gyu , Kim Dong-Hoon , Kim Dong-Kyo , Kim Hyun-Min , No Jin-Gu , Yeom Dong-Hyeon , Choi Mi-Kyung , Nam Yoon-Seok

Introduction: Accurate measurement of ovulation timing is difficult in the dog. In this study, we investigate the expression of steroid hormone receptors in the ovary and oviduct before and after ovulation in the dog.Material and methods: Estrus and ovulation period were measured by serum concentrations of progesterone hormone and LH. LH surge was defined as day 0. The ovary and oviduct were isolated at the day before LH surge (day-1), 2 day after ovulat...

ra0001p137 | (1) | WCRB2014

Effect of progesterone levels and parity on the maturity of in vivo canine oocytes

Kim Dong-Hoon , Choi Mi-Kyung , No Jin-Gu , Yeom Dong-Hyeon , Kim Hyun-Min , Kim Dong-Kyo , Yang Byoung-Chul , Yoo Jae Gyu

Somatic cell nuclear transfer (SCNT) in dogs has been used in vivo-matured oocytes due to low efficiency of in vitro maturation system. Actually, more information is needed to acquire good quality in vivo oocytes for the dog research. The objective of present study was to evaluate whether the progesterone (P4) on estimated ovulation time and the parity of donor bitches have any effect on the maturity of oocytes. In vivo oocytes were retrieve...

ra0001p138 | (1) | WCRB2014

The development of a novel mouse embryonic ovary culture

Stefansdottir Agnes , Adams Ian , Spears Norah

Introduction: In vitro cultures are a widely used tool to study ovary development and assess reproductive toxicology of chemicals. However, establishing a culture system whereby mouse ovaries can be cultured from a pre-meiotic stage to a mature oocyte has proved challenging. We have developed a novel culture system that spans meiotic entry to meiotic arrest, germ cell nest break-down, follicle formation, and initiation of follicle growth.Methods...

ra0001p139 | (1) | WCRB2014

Regulatory role of resistin on ovarian function

Rak-Mardyla Agnieszka , Wrobel Anna , Drwal Eliza , Gregoraszczuk Ewa

Introduction: We recently reported expression of resistin in porcine ovarian follicles and stimulatory on basal why inhibitory on gonadotropin-stimulated ovarian steroidogenesis by direct influence on 3β-HSD and 17β-HSD expression. The question is whether resistin, beyond the control of steroidogenesis, regulates apoptosis in ovarian follicles. The present in vitro study was designed to asses effect of resistin on basal and FSH or IGF1 stimulated caspases ac...

ra0001p140 | (1) | WCRB2014

The effects of bisphenol A in in vitro neonatal rat ovary

Altunbas Korhan , Yagci Artay , Celik Sefa , Akkaya Ozlem Ozden , Zik Berrin

Introduction: Neonatal bisphenol A (BPA) exposures affect the developing reproductive tract of females, several distinct stages of oogenesis in the developing ovary. Notch signaling also plays an important role during oogenesis. We aimed to explain whether mRNA levels of Notch receptor and ligands are altered following BPA treatment during neonatal days in vitro.Materials and methods: Wistar rats were sacrificed and ovaries were isolated on post...

ra0001p141 | (1) | WCRB2014

Acquisition of meiotic competence in porcine secondary follicles following long-term culture

Tasaki Hidetaka , Itami Nobuhiko , Iwata Hisataka

The development of culture methods to produce oocytes from immature ovarian follicles in vitro would greatly contribute to livestock production and assisted human reproduction. However, successful in vitro culture to produce fully mature oocytes is very difficult in the case of large domestic animals. In the present study, we investigated the growth properties of in vitro-cultured porcine oocytes derived from early developmental stage follicles. Oocy...

ra0001p142 | (1) | WCRB2014

Isolation of primordial and primary follicles from porcine ovarian medulla tissue

Duda Malgorzata , Grzesiak Malgorzata , Tabarowski Zbigniew , Tomanek Milan

Introduction: The ovarian follicle has a three-dimensional structure in which the oocyte is surrounded by tightly connected follicle cells that mediate the action of external signals and nourish the gamete during its maturation. In vitro follicle techniques provide a tool to model follicle development in order to investigate basic biology and are further being developed as a technique to preserve human fertility or to product transgenic animals for cell therapies. Mai...

ra0001p143 | (1) | WCRB2014

Analysis of programmed cell death of porcine granulosa cells treated with vinclozolin

Slomczynska Maria , Knet Malgorzata , Tabarowski Zbigniew , Duda Malgorzata

Introduction: Androgens: testosterone, dihydrotestosterone (DHT), and a fungicide with antiandrogenic activity – vinclozolin (Vnz) served to study agonism and antagonism of the androgen receptor.Material and methods: Granulosa cells (GC) isolated from pig follicles were cultured for 48 h with testosterone (10−7 M), DHT (10−7 M), and Vnz (1.4×10−5 M) separately or in combinations. Cells were the...

ra0001p144 | (1) | WCRB2014

Etoposide has a detrimental impact on mouse ovarian development when exposure occurs during early meiotic prophase

Johnston Zoe C , Stefansdottir Agnes , Adams Ian , Spears Norah

Introduction: The use of the chemotherapeutic agent etoposide in pregnancy is considered to be relatively safe during the second and third trimesters. However, the drug does have detrimental effects on oocytes undergoing meiosis II. Similar effects on oocytes in meiosis I may have a clinical impact on the fertility of women exposed to the drug in utero, during critical stages of ovarian development. This study aims to examine the effects of etoposide exposure during e...

ra0001p145 | (1) | WCRB2014

The age-associated deterioration in the follicular fluid induces a decline in bovine oocyte quality

Takeo Shun , Kuwayama Takehito , Iwata Hisataka

Introduction: In general, the maternal age affects the quality of the oocytes as well as that of embryos. This study aims to examine whether changes in the follicular fluid (FF) can affect oocyte quality in aged cows.Materials and methods: Oocytes were collected from ovaries of young (<120 months) or aged cows (≥120 months), and oocytes from animals in these two age groups were cultured in a maturation medium that contained 10% FF derived from ...

ra0001p146 | (1) | WCRB2014

Resveratrol enhances the in vitro development of oocytes derived from early antral follicles

Itami Nobuhiko , Kuwayama Takehito , Iwata Hisataka

Introduction: The present study aims to investigate the effect of resveratrol supplementation on the in vitro development of porcine oocytes derived from early antral follicles.Materials and methods: Oocyte-granulosa cell complexes (OGCs) were collected from early antral follicles that were 0.5–0.7 mm in diameter and cultured in the absence or present of resveratrol (2 μM) for 14 days. The effect of resveratrol on SIRT1 expression was ...

ra0001p147 | (1) | WCRB2014

Expression of pentraxin 3 transcript and protein in porcine preovulatory follicles: comparison of in vivo and in vitro conditions

Nagyova Eva , Nemcova Lucie , Kalous Jaroslav , Salustri Antonietta , Camaioni Antonella

It has been previously shown that multimeric pentraxin 3 (PTX3) is a key component of the cumulus oophorus extracellular matrix. Noteworthy, cumuli from PTX3−/− mice were defective in matrix organization and fertility was impaired. PTX3 binds to both tumor necrosis factor alpha-induced protein 6 (TNFAIP6) and inter-alpha-trypsin inhibitor thereby likely serving as a cross linker for multiple hyaluronan molecules, stabilizing in this way the cumulus oopho...

ra0001p148 | (1) | WCRB2014

Cell viability in ovarian follicles exposed to cisplatin and doxorubicin

Kirkwood Phoebe Maud , Lopes Federica , Spears Norah

Introduction: This work adapts a novel ovary culture technique, devised by De Felici’s group, for assessing the ovotoxicity of two chemotherapeutic drugs, cisplatin and doxorubicin. Previous work from our laboratory used cultures of intact neonatal ovaries to determine the effects of the two drugs, showing that both cisplatin and doxorubicin are moderately ovotoxic.Methods: Here, ovaries collected from newborn WT mice were cultured as by De Felici. ...

ra0001p149 | (1) | WCRB2014

Ghrelin, like adipokines produced by adipose tissue, acts on accelerate puberty, while in contrast to adipokines, during sexual maturity prevents from excessive secretion of steroids by ovarian follicles

Rak-Mardyla Agnieszka , Wrobel Anna , Gregoraszczuk Ewa

Introduction: In our previously published study we focused on actions of ghrelin on ovarian steroidogenesis in prepubertal period and indicated that as adipokines produced by adipose tissue (such as leptin or resistin), ghrelin acts on accelerate puberty. This data had been performing to determine whether also in cycling animals ghrelin acts on ovarian steroidogenesis similarly to leptin and resistin.Material and methods: Small (SFs), medium (MFs), and l...

ra0001p150 | (1) | WCRB2014

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ra0001p151 | (1) | WCRB2014

Detection of ZP-autoantibody in the sera from infertile women using a human multiepitope peptide as antigen

Xu Wan Xiang , Wang Jian , Zhu Weijie , Chang Xiaoxue , Sun Xiaoxi , Gupta Satish K , He Yaping , Tang Haiping , Shi Huijuan , Gu Shaohua , Ji Chaoneng , Xie Yi

Background: The role of autoantibodies against zona pellucida (ZP) in infertility and success of IVF is debatable. In the present study, a novel recombinant human zona protein is used to address this issue.Materials and methods: Fourteen serum samples positive for porcine ZP, six from infertile patients, and 20 from healthy women were analysed. Sera were tested for their reactivity against a recombinant multi-epitope peptide (named ZPCP7) composed of ten...

ra0001p152 | (1) | WCRB2014

Occurrence of rare three types of chromosome configurations in a Murrah buffalo (Bubalus Bubalis)

Sen Sulochana , Tonk Rakesh , Yadav B R

Introduction: Chromosomal abnormalities are deviations in normal genetic architecture and lead to disorders in bearer individuals. These can be both in number and structure of autosomes or sex chromosomes, usually inflict adverse effects on reproductive performance in domestic animals. Present report is on a female Murrah buffalo (Bubalus bubalis), which was among animals with inefficient reproductive performance and cytogenetic screening programme.<p class="abste...

ra0001p153 | (1) | WCRB2014

Effect of different diet fat sources on in vitro maturation of Anglo-Nubian goats oocytes

Nociti Ricardo P , Padilha Luciana C , Rola Luciana D , Barros Felipe F F P C , Uscategui Ricardo R , Mariano Renata S G , Feliciano Marcus A R , Vicente Wilter R R , Oliveira Maria Emilia F , Lima Vera F M H

There is a lack of information about effects of the diet fat source on goat reproduction. The aim of the present study was to evaluate the fat source effect on in vitro maturation (IVM) of oocytes. Eighteen goats were divided into three groups, on diets (40% concentrate/60% corn silage) with 4% (dry matter) of ether extract from different fat sources [soy oil (SG) and linseed (LG), Megalac® (MG)]. The does passed thru an adaptation period (15 days) and...

ra0001p154 | (1) | WCRB2014

Chronic sympathetic stress during gestation delays puberty and follicular development of female offspring

Barra Rafael , Cruz Gonzalo , Mayerhofer Artur , Paredes Alfonso , Lara Hernan E

It is widely accepted in the literature that the exposure to stress during early stages of development could have permanent and severe effect in the developing organism. We studied the effect of maternal sympathetic stress to pregnant rats, and studied the permanent effects in follicular development of the ovary of the progeny.Methods: Pregnant Sprague – Dawley rats were cold stressed (4 °C 3 h/day) during all pregnancy. Control group was maint...

ra0001p155 | (1) | WCRB2014

Proteins in porcine follicular fluid as potential biomarkers for fertility

Jarrett Selene , Gill Andy C , Kurian Dominic , Hogg Charis O , Ferguson Elizabeth M , Ashworth Cheryl Joy

Introduction: Following IVF, blastocysts produced from oocytes recovered from gilts fed a high fibre diet for the preceding 19 days contained more cells than blastocysts from oocytes of control-fed gilts. Oocytes were collected on day 19 of the gilts’ third oestrous cycle and matured in 10% of their own follicular fluid (FF), suggesting that FF may confer the reproductive benefits. The current study compared the protein composition of pooled FF from six high fibre-fed pig...

ra0001p156 | (1) | WCRB2014

Glucocorticoid metabolism in bovine cumulus–oocyte complex during in vitro maturation

Tetsuka Masafumi , Takagi Ryo , Ambo Nobuhiro , Zempo Yuta , Onuma Asuka

Introduction: Tone of glucocorticoid action in target organs is regulated by relative activity of 11βHSD type1 (11HSD1), a bidirectional oxidoreductase that mainly converts cortisone to active cortisol, and type 2 (11HSD2), an oxidase that inactivates cortisol to cortisone. We have demonstrated that bovine cumulus–oocyte complex (COC) expresses 11HSDs and both reductive and oxidative activities are present in COC undergoing IVM. In the present study we investigated r...

ra0001p157 | (1) | WCRB2014

A high concentration of polyvinylpyrrolidone in culture medium affects the number of tranzonal projections in bovine oocyte–granulosa cell complexes in long-term culture

Hirao Yuji , Somfai Tamas , Akagi Satoshi , Haraguchi Seiki , Watanabe Shinya

Introduction: A high concentration of polyvinylpyrrolidone (PVP, average molecular weight 360 000) supplementation in culture medium exerts a positive influence on the growth and morphological organization of bovine oocyte–cumulus/granulosa cell complexes in a long-term culture. The objective of the present study was to determine the effect of PVP supplementation on the number of transzonal projections (TZPs) traversing the zona pellucida from cumulus cells to the oocyte....

ra0001p158 | (1) | WCRB2014

Mouse CD11c+ dendritic cells are crucial in folliculogenesis, luteinization, and luteal maintenance

Nagai Miwako , Koga Kaori

Introduction: The ovary shows cyclical events, such as folliculogenesis, ovulation, and luteinization. During these events, leukocytes are known to infiltrate in the ovary. However, the role of each type of leukocytes in the ovary is not fully determined. In order to determine the roles of dendritic cells (DC) in the ovary, we conducted following experiments.Materials and methods: i) To analyze DC population, immature mice were injected with PMSG followe...

ra0001p159 | (1) | WCRB2014

Investigating whether Kiss1 KO mice can be used as a model for PCOS and age-onset diabetes

Kyle Victoria , d'Anglemont de Tassigny Xavier , Colledge William H

Polycystic ovary syndrome (PCOS) is generally thought to be a genetic disease, which affects 5–10% of women of reproductive age (approximately 11–47 years old). Typically, PCOS is characterized by the formation of cysts on the ovaries and ovulatory failure. Characteristically PCOS results in high levels of androgens such as testosterone and insulin resistance within the blood. The ovaries of Kiss1 mutant mice have been shown to frequently form multiple cysts...

ra0001p160 | (1) | WCRB2014

Androgen action within the preantral follicle; a role for the epidermal growth factor receptor?

Thomson Kacie , Laird Mhairi , Franks Stephen , Hardy Kate

Introduction: Polycystic ovary syndrome (PCOS) is the most common cause of anovulatory infertility affecting over 5% of the female population. PCOS is characterized by aberrant early follicle development in which hyperandrogenism is thought to play a key role. However, the molecular mechanisms of androgen action within the ovary remain largely unknown. Recent evidence suggests androgens may be acting, in part, through modulation of growth factor signalling. We have recently sh...

ra0001p161 | (1) | WCRB2014

Differential role of TGFβ2 during follicle activation and preantral follicle growth

Oliver Elizabeth , Laird Mhairi , Franks Stephen , Hardy Kate

Introduction: Female mammals are born with a finite reserve of primordial follicles which dictates the reproductive lifespan. Despite this the factors responsible for primordial follicle activation remain largely unknown. The transforming growth factor beta (TGFβ) superfamily has been implicated in early follicle development however little attention has been given to the effects of TGFβ2, expression of which has been demonstrated in the ovary. This study aimed to exa...

ra0001p162 | (1) | WCRB2014

Mycotoxin-contaminated diet affects mouse oocyte quality

Hou Yan-Jun , Zhu Cheng-Cheng , Cui Xiang-Shun , Kim Nam-Hyung , Sun Shao-Chen

Introduction: Mycotoxins, such as deoxynivalenol (DON), zearalenone (ZEN), and aflatoxin (AF), are commonly found in many food commodities and may impair the growth and reproductive efficiency of animals and humans. We investigated the effects of a mycotoxin-contaminated diet on mouse oocyte quality.Materials and methods: Maize contaminated with DON (3.875 mg/kg), ZEN (1,897 μg/kg), and AF (806 μg/kg) was incorporated into a mouse diet at three...

ra0001p163 | (1) | WCRB2014

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ra0001p164 | (1) | WCRB2014

Obesity depresses Toll-like receptor superfamily members in the ovary

Galvao Antonio , Socha Barbara , Jura Magdalena , Nowialis Pawel , Skarzynski Dariusz

Introduction: Obesity and its associated metabolic disorders lead to ovarian failure and infertility. Nonetheless, the hierarchy of pathways leading to ovarian dysfunction is unknown. Presently, we studied the impact of obesity on ovarian innate immune response and steroidogenesis throughout the time.Materials and methods: An in vivo study was conducted on C57Bl/6J mice (n=8/group) fed a chow-diet versus high-fat diet during 4- (C-4w; H...

ra0001p165 | (1) | WCRB2014

Space pup project: sperm preservation in space station

Wakayama Sayaka , Yano Sachiko , Kasahara Haruo , Osada Ikuko , Shimizu Toru , Suzui Hiromi , Mizutani Eiji

Sustaining life beyond earth either on space stations or other planets will require a clear understanding of how the environment in space affects key phases of mammalian reproduction. However, because of the difficulty of conducting such experiments in live animals, most studies on reproduction in space have thus far been carried out in species such as fish or amphibians. Mammalian embryos can only be cultured for a few days and could pose technical challenges for the astronau...

ra0001p166 | (1) | WCRB2014

Amino acid and glucose profiles of bovine oviduct epithelial cell secretions in response to 17β-oestradiol and progesterone

Simintiras Constantine A , Sathyapalan Thozhukat , Leese Henry J , Sturmey Roger

Oviduct luminal fluid supports early embryo development. The composition this fluid has been debated despite studies using post mortem samples and direct sampling under anesthesia. The routine availability of such data would benefit in vitro embryo culture in addition to our understanding of the influences of maternal physiology on in vivo oviduct fluid composition.We have devised an in vitro model for the determination of bovi...

ra0001p167 | (1) | WCRB2014

Generation of rats from vitrified oocytes with surrounding cumulus cells via IVF with cryopreserved sperm

Kashiwazaki Naomi , Fujiwara Katsuyoshi , Kato Tsubasa , Kamoshita M , Obata H , Takakusa Syouta , Ito Junya

The aim of the present study was to evaluate fertility of vitrified rat matured-oocytes with surrounding cumulus cells after IVF with cryopreserved sperm. Oocytes with surrounding cumulus cells and cumulus cells denuded oocytes were vitrified with 30% (v/v) ethylene glycol+0.5 M sucrose+20% (v/v) FCS in PBS (PB1) by using Cryotop. After warming, oocytes were co-cultured with fresh or cryopreserved epididymal sperm for 10 h. Although the fertilization (two pronuclei formation: ...

ra0001p168 | (1) | WCRB2014

Comparative effects of administration of kisspeptin-10 and GnRH on LH secretion in buffalo cows

Chaikhun-Marcou Thuchadaporn , Sotthibandhu Pongsiwa , Suthikrai Wanwipa , Jintana Ratree , Makoom Pussadee , Suadsong Siriwat

Introduction: Previous studies have proven the efficiency of GnRH for stimulating both the pulse and amplitude of LH releasing in buffaloes, but not study of kisspeptin efficiency.Materials and methods: There were three treatments: i) kisspeptin-10 (K-10; a single i.v. of 1000 pmol/kg b.w. dosage), ii) GnRH (a single i.m. injection of 10 μg/cow dosage), and iii) distilled water (DW; a 2 ml single i.v. injection). All six animals were treated and the...

ra0001p169 | (1) | WCRB2014

Ovulation induction in prepubertal Sahiwal×Friesian cross bred heifers by the use of clomiphene citrate and hCG

-ur-Rehman Ubaid , Ali Shujait , Ahmad Nazir , Khanum Shahnaz Adeeb

Introduction: Clomiphene citrate belongs to group of drugs known as selective estrogen receptor modulators. In this study, an attempt was made to induce ovulation in crossbred prepubertal heifers by the use of clomiphene citrate and hCG. It was hypothesized that clomiphene citrate, being antiestrogen, would remove negative feedback effect of estrogen on the release of pituitary gonadotropins. Elevated gonadotropins would support follicular growth to a point where hCG would ind...

ra0001p170 | (1) | WCRB2014

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ra0001p171 | (1) | WCRB2014

Evaluation of the cyclophosphamide gonadotoxicity and the protective effect of GnRH analogues in mice model

Horicks Florence , Bockstaele Laurence , Houben Sarah , Van den Steen Geraldine , Englert Yvon , Demeestere Isabelle

Introduction: While gonadotoxicity of cyclophosphamide (Cy) has been well demonstrated in mice model, the kinetic and effect at different follicular stages are still not well described. Understanding these phenomena is however essential to further develop pharmacological protective approaches. Here, we studied the effect of Cy at different doses and timing on follicular development as well as the potential gonadoprotective effect of the GnRH analogues (GnRHa) during chemothera...

ra0001p172 | (1) | WCRB2014

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ra0001p173 | (1) | WCRB2014

Fibrotic remodeling and duct obstruction are major patho-physiological mechanisms in bacterial epididymitis in mice

Michel Vera , Bhushan Sudhanshu , Middendorff Ralf , Loveland Kate L , de Kretser David D , Hedger Mark P , Meinhardt Andreas

Introduction: Bacterial epididymitis is a common disorder in urological outpatient clinics and usually results from ascending infections originating in the urinary tract often involving uropathogenic E. coli (UPEC) as causative pathogen. In 40% of patients with acute epididymitis, impaired semen parameters persist even after antibiotic treatment. Members of the TGFβ cytokine superfamily, such as activin A, have recently been identified in the epididymis and are c...

ra0001p174 | (1) | WCRB2014

Model for differentiation of mouse embryonic stem cells into steroidogenic cells

Kang Hee Young , Lee Jae-Hwan , Jeung Eui-Bae

Introduction: Steroidogenic factor 1 (SF-1) is essential for the development and function of steroidogenic tissues. Stable incorporation of SF-1 into embryonic stem cells has been reported to prime the cells for steroidogenesis.Materials and methods: In this study, we established SF1 transgenic mouse embryonic stem cell (SF1-mES cells) and analyzed expression of steroidogenesis-related genes and gonadal lineage-markers. We measured the secreted progester...

ra0001p175 | (1) | WCRB2014

Early life programming of adult Leydig cell function

Kilcoyne Karen R , Smith Lee , Jobling Matthew , McKinnell Chris , MacPherson Sheila , van den Driesche Sander , De Gendt Karel , Verhoeven Guido , Anderson Richard , Sharpe Richard

Introduction: Adult male testosterone levels are influenced by fetal events, but how is unknown, as adult Leydig cells (ALC) do not differentiate until puberty. We hypothesised that adult Leydig stem cells are present in the fetal testis, and are susceptible to programming by fetal androgen exposure. We have shown that ALC derive from interstitial cells that express chicken ovalbumin upstream promoter transcription-factor II, essential postnatally for ALC development, and andr...

ra0001p176 | (1) | WCRB2014

Characterization of pituitary-derived cell lines, Tpit/E, Tpit/F1 and TtT/GF

Nishimura Naoto , Yoshida Saishu , Higuchi Masashi , Yako Hideji , Ueharu Hiroki , Chen Mo , Kato Takako , Kato Yukio

Introduction: The pituitary anterior lobe consists of five types of hormone-producing cells and non-hormonal cells. In non-hormonal cells, Sox2-expressing cells exist as stem/progenitor cells and play a role in the regeneration of hormonal cells. However, elucidation of the differentiation mechanism is yet uncovered. Established cell lines are frequently used as a model system not only for cellular physiology and gene expression but also for cell differentiation. In t...

ra0001p177 | (1) | WCRB2014

Functional and molecular features of the ID4+ germline stem cell population in mouse testes

Oatley Melissa , Chan Frieda , Kaucher Amy , Oatley Jon

Continual spermatogenesis relies on the actions of an undifferentiated spermatogonial population that is comprised of stem cell and progenitor fractions. Self-renewal by spermatogonial stem cells (SSCs) maintains a foundational pool from which progenitor spermatogonia arise that transiently amplify in number before committing to a pathway of terminal differentiation. At present, molecular features that distinguish the stem cell state in mammalian spermatogonia are undefined. I...

ra0001p178 | (1) | WCRB2014

Presence of the juxtacrine factor EphrinB2 in a rat pituitary stem/progenitor cell niche

Yoshida Saishu , Kawai Kohei , Kato Takako , Kato Yukio

Introduction: The anterior pituitary lobe is a key endocrine tissue composed of five types of endocrine-cells and non-hormonal cells. Among non-endocrine-cells, Sox2-expressing cells exist as stem/progenitor cells and play a role in the regeneration of endocrine-cells in the adult pituitary. Recently, we have reported that transcription factors, Prop1, Prrx1 and Prrx2, are expressed in the pituitary stem/progenitor cells and these cells cont...

ra0001p179 | (1) | WCRB2014

Krüppel-like factor 6 (klf6) is expressed in rat pituitary stem/progenitor cells and regulates the PRRX2 gene

Ueharu Hiroki , Higuchi Masashi , Nishimura Naoto , Yoshida Saishu , Nishihara Hiroto , Kato Takako , Kato Yukio

Introduction: Paired-related transcription factors, PRRX1 and PRRX2, which are expressed in mesenchymal tissues and participate in mesenchymal cell differentiation, were recently found in stem/progenitor cells of the pituitary gland of ectodermal origin. The present study aimed to clarify the role of PRRX1 and PRRX2 in the pituitary gland by identifying factors that regulate the expression of both genes.Materials and methods: Immunohistochemistry, promot...

ra0001p180 | (1) | WCRB2014

Concentration levels of TSH evaluation with species-specific MABs due to the dog reproduction disorders assessment in the course of hypothyroidism

Aniolek Olga , Gajewski Zdzislaw , Domino Malgorzata

Introduction: Hypothyroidism is the disorder influences on reproduction of many species, including dogs. However the results are different, even among the same species.Materials and methods: The concentration of TSH with species-specific monoclonal anti-TSH in dogs hypothyroidism was evaluated. TSH concentration was measured with a chemiluminescent ELISA solid phase two-point (TSH canine Immulite, DPC, LA, CA). Coefficients of variation within one were 5...

ra0001p181 | (1) | WCRB2014

Invasion of S100β-positive cells into pituitary gland during embryonic period

Yako Hideji , Horiguchi Kotaro , Higuchi Masashi , Fujiwara Ken , Yoshida Saishu , Chen Mo , Naoko Kanno , Ueharu Hiroki , Kato Takako , Yashiro Takashi , Kato Yukio

Introduction: Pituitary cells positive for S100 protein are non-hormone-producing cells and are composed of heterogeneous populations, including folliculo-stellate cell. They are argued for having various functions and are detectable only after birth, but their origin is not yet clarified. Recently, we had detected S100βmRNA in the cDNA library of embryonic pituitary by real-time PCR and observed invasion of non-hormone-producing cells from surrounding mesenchyme...

ra0001p182 | (1) | WCRB2014

Neuronatin is first expressed in pituitary stem/progenitor cells

Naoko Kanno , Higuchi Masashi , Yako Hideji , Yoshida Saishu , Kato Takako , Chen Mo , Kato Yukio

Introduction: Neuronatin (NNAT) is first identified in the neonatal neural tissue and is assumed to be involved in embryonic neurogenesis. Although defect of PROP1, a pituitary specific transcription factor important for pituitary organogenesis, causes reduced expression of the Nnat, the cellular localization and the vital roles of NNAT in the pituitary are still unclear. The present study examined the population of the NNAT-positive cells as to understand the role of NNAT in ...

ra0001p183 | (1) | WCRB2014

Early aging of mesenchymal stem/stromal cells derived from the bone marrow of α-1,3-galactosyltransferase knockout pig

Ock Sun A , Oh Keon Bong , Hwang Seongsoo , Im Seo Ki , Kim Youngim

Introduction: The α-1,3-galactosyltransferase knockout (GalT KO) pig is used as the major model for xenotransplantation studies since the knockout of the α-1,3-galactosyl epitope makes pig organ transplantation successful in primates. The homozygous GalT KO pig is produced by a combination of nuclear transfer technique and mating with a heterozygous GalT KO pig. Since most studies using GalT KO pigs have analyzed only the immunological features of their cells, we...

ra0001p184 | (1) | WCRB2014

Derivation of naive-type induced pluripotent stem cells in cattle using piggyBac transposition of doxycycline-inducible transcription factors

Kawaguchi Takamasa , Tsukiyama Tomoyuki , Minami Naojiro , Yamada Masayasu , Matsuyama Shuichi , Kimura Koji , Imai Hiroshi

Pluripotent stem cells (PSCs) in large domestic animals including cattle are expected to be used as means for genetic improvement and production of disease-model animals. There are two types of PSCs: naive and primed. Ontogenetically, naive PSCs correspond to an earlier developmental state compared to other types of PSCs and have the capacity for producing chimeric offspring. Previously reported PSCs in large domestic animals are classified as primed type. In this study, putat...

ra0001p185 | (1) | WCRB2014

Serum-free culture of mouse trophoblast stem cells

Sun Shuai , Yano Shota , Nakanishi Momo O , Hirose Michiko , Nakabayashi Kazuhiko , Hata Kenichiro , Ogura Atsuo , Shiota Kunio , Tanaka Satoshi

Introduction: Trophoblast stem cells (TSC) maintain their undifferentiated status under the presence of fibroblast growth factor 4 (FGF4), heparin and feeder cell-conditioned medium (CM). It has been reported that activin A can replace CM. However, even in such stem cell condition, a portion of TSC spontaneously undergoes differentiation. In addition, when TSC are induced to differentiate, all trophoblast subtypes autonomously appear without the addition of any exogenous facto...

ra0001p186 | (1) | WCRB2014

Incomplete reprogramming of bovine fetal fibroblasts by the stimulus of low pH*

Jin Sang Jin , Kim Tae Suk , Baek Sang Ki , Moon Song Yi , Ko Yeoung-Gyu , Kim Sung Woo , Seong Hwan Hoo , Lee Joon Hee

Stem cells can provide a useful tool for studying on mechanisms of embryonic development and production of transgenic animal. Although somatic cells are able to convert into reprogrammed cells by introduction of exogenous reprogramming factors, however, the efficiency of iPSC is still low. Murine somatic cells without transfection of exogenous transcription factors were easily able to be reprogrammed by stimulus of low pH. Here we attempted whether to convert bFFs into pluripo...

ra0001p187 | (1) | WCRB2014

Leukemia inhibitor factor (LIF) is essential for long term maintenance of pluripotency of porcine induced pluripotent stem cells*

Baek Sang Ki , Kim Tae Suk , Moon Song Yi , Jin Sang Jin , Ko Yeoung Gyu , Kim Sung Woo , Seong Hwan Hoo , Lee Joon Hee

piPS cells are divided into naïve and primed states. These states may depend on culture conditions with/without cytokines. Only piPS cells of naïve state have the capacity for producing chimeric offspring and long term maintenance of pluripotency. The objective of this study was to generate piPS cells of naïve state under culture conditions with 2i of the Erk/2 and of GSK3. PFFs were transduced with SIX reprogramming factors. After the transduction piPS cells we...

ra0001p188 | (1) | WCRB2014

Function of leukemia inhibitory factor in spermatogenesis, revealed by using medaka spermatogonial culture system

Satoh Ryuichi , Yamashita Masakane

Introduction: Spermatogenesis is a dynamic process. Its control mechanisms are very complex and many molecules are concerned. Our previous study showed that spermatogonia are abnormally proliferated when p53 (a tumor-suppressor gene) was knocked out in medaka (p53-KO medaka). The p53-KO medaka provides a hint about the regulatory mechanisms of spermatogonial proliferation and differentiation.Materials and Methods: Since some cy...

ra0001p189 | (1) | WCRB2014

RNA binding protein Lin28a interacts with various mRNA in the mouse trophoblast stem cells

Sakamaki Yuji , Hayakawa Koji , Tanaka Satoshi

Introduction: Trophoblast stem (TS) cells are capable of differentiating into all subtypes of trophoblast cells. Lin28a, an RNA-binding protein involved in the post-transcriptional regulation of gene expression, is highly expressed in undifferentiated TS cells but is greatly reduced in its expression in differentiated trophoblast cells. Lin28a is known as a major factor controlling growth and metabolism through regulation of the maturation of let-7 microRNA. Consideri...

ra0001p190 | (1) | WCRB2014

Derivation of porcine iPS-like cells from fibroblast of a translocated azoospermic boar

Congras Annabelle , Barasc Harmonie , Vignoles Florence , Delcros Chantal , Pinton Alain , Canale-Tabet Kamila , Feraud Olivier , Turhan Ali , Yerle-Bouissou Martine , Acloque Herve

Chromosomal rearrangements have a crucial impact on the proper proceedings of meiosis and can lead by several mechanisms to the production of unbalanced gametes or to the complete arrest of gametes production. To assess the impact of these rearrangement in the early development of pig germ cells, we proposed to generate a library of stem cells from an azoospermic boar carrying a reciprocal translocation t(Y:14), as a new tool for the development of an in vitro differe...

ra0001p191 | (1) | WCRB2014

Derivation of bovine induced pluripotent stem cells by a transposon approach

Rao Talluri Thirumala , Kumar Dharmendra , Glage Silke , Garrels Wiebke , Debowski Katharina , Behr Rudiger , Niemann Heiner , Kues Wilfried

Introduction: Induced pluripotent stem (iPS) cells are considered as a seminal breakthrough in stem cell research and are promising for the development of advanced regenerative therapies. Considering the potential of this technology for both basic and clinical research, it is tempting to extend this research to important livestock species, such as cattle, in which ES cell lines are yet not available.Materials and methods: Here, we describe a non-viral me...

ra0001p192 | (1) | WCRB2014

Effect of melatonin supplementation during in vitro maturation on intracellular ROS levels in porcine oocytes

Cruz Maria Helena Coelho , Cruz Jurandir Ferreira , Reiter Russel J , Schwarz Katia Lancellotti , Castro Fernanda Cavallari , Cunha Maria Carolina R V , Verde Leal Claudia Lima

Oocytes of domestic mammals present high concentrations of lipids, and this is particularly evident in porcine oocytes that exhibit higher levels than other species, becoming more susceptible to oxidative stress and lipid peroxidation. Considering that melatonin is an effective antioxidant for protecting macromolecules against oxidative stress caused by reactive oxygen species (ROS), the purpose of the present study was to examine the effect of melatonin supplementation during...

ra0001p193 | (1) | WCRB2014

X chromosome dosage affects completion of pluripotential reprogramming

Kim Jong Soo , Choi Hyun Woo , Hong Yean Ju , Scholer Hans R , Do Jeong Tae

Direct reprogramming of somatic cells to pluripotent stem cells entails the obliteration of somatic cell memory and the reestablishment of epigenetic events. Induced pluripotent stem (iPS) cells have been created by reprogramming somatic cells through the transduction of reprogramming factors. During cell reprogramming, female somatic cells must overcome at least one more barrier to enter a pluripotent state compared to male somatic cells, that is, the reactivation of inactive...

ra0001p194 | (1) | WCRB2014

High-throughput screening revealed a clinically relevant drug to induce sperm motility

Cochrane Clair , Ruso Halil , Hope Anthony , Clarke Rosemary G , Barratt Christopher , da Silva Sarah Martins

Introduction: Sperm dysfunction is the commonest cause of infertility, yet there is currently no drug a man can take, or be added to his sperm in-vitro, to improve fertility. The rationale of drug discovery research is to find a drug which increases sperm motility and success of ART. Despite limitations in understanding of sperm physiology, it is acknowledged calcium is central to motility and function. Validated high-throughput screening of compounds from University ...

ra0001p195 | (1) | WCRB2014

Steps forward canine primordial germ cell isolation and characterization

de Souza Aline Fernanda , Pieri Naira Caroline Godoy , Cassals Juliana Barbosa , Mancanares Ana Carolina Furlanetto , Roballo Kelly Cristine Santos , Ambrosio Carlos Eduardo , Perecin Felipe , Martins Daniele Santos

Introduction: Primordial germ cells (PGCs) are specified during early mammalian post implantation development. They are precursors of adult animals gametes and are considered stem cells of germline. Most of PGCs studies were performed in rodents or human with limited studies in other mammals such as canine. We have recently demonstrated a model to derive embryonic germ cells (EGC) in canine species.Materials and methods: We isolated canine embryos betwee...

ra0001p196 | (1) | WCRB2014

Effects of early embryo environment on embryonic stem cell derivation and behaviour

Cox Andrew L , Sun Congshan , Sheth Bhav , Fleming Tom , Smyth Neil

The embryonic environment can induce permanent changes in metabolism and increase disease risk in adults. Rodent models show that modifications in maternal diet exclusively during pre-implantation development can induce metabolic disease in adults. Changes must therefore occur within the early embryo and be maintained throughout development. Determining adaptive mechanisms is challenging due to the size of the early embryo. We generated ES cells from inbred C57BL/6 mice as a m...

ra0001p197 | (1) | WCRB2014

Trophectoderm stem cells to model the effect of altered periconceptional diet on embryos

Chang Yi-Lung , Fleming Tom , Smyth Neil

Introduction: Poor maternal diet during development induces physiological, metabolic and anatomical alterations in offspring. These may increase immediate survival chances, but can enhance the risk of disease in later life. Maternal low protein diet (LPD) exclusively in pre-implantation development induces changes in endocytosis, postnatal hypertension, cardiovascular dysfunction and alters behaviour in our mouse model. We have previously shown changes in the embryonic stem ce...

ra0001p198 | (1) | WCRB2014

Effect of storage time on Atlantic salmon (Salmo salar) sperm motility, viability and calcium levels, first communication

Parodi Jorge

Few studies have evaluated sperm function and intracellular calcium levels when storage of semen for a long period of time. We have assessed sperm viability, morphology, motility and intracellular calcium levels in fish semen after a prolonged storage period. Semen samples from Atlantic salmon were preserved at 4 °C for 25 days and then evaluated for viability. The motile sperm was determined using the Image J-CASA program. To measure the calcium content, the sperm were l...

ra0001p199 | (1) | WCRB2014

Motility, viability and calcium in the sperm cells

Parodi Jorge

Sperm cells are complicated in vitro models. Its viability is limited, and its physiology is complex. The study of their properties is of great application in the animal production industry. Therefore, it is important to have viable and functional gametes. Consequently, it has been demonstrated that the decrease of sperm cells viability is related with the increase of the reactive oxygen species (ROS) and ROS is secondary to normal metabolic processes of the cell. One...

ra0001p200 | (1) | WCRB2014

Reciprocity of bio-stimulation on testicular traits and subsequent fertility of South African Mukota sows

Umesiobi Dennis Onuoha

Introduction: Bio-stimulation is the stimulus provoked by the presence of a male which induces fertility through genital stimulation or other external cues. In animals, courtship and foreplay which precede mating have reciprocal effects on sexual and reproductive milieu of both sexes. An increase in male exposure to females will also tend to evoke increases in reproductive capacity of both sexes. The study evaluated reciprocity of boar exposure on testicular characteristics an...

ra0001p201 | (1) | WCRB2014

Alternative splicing: a mechanism for spermatogonia differentiation, meiosis progression and spermatid maturation

Jamsai Duangporn , Clark Brett , McLaughlin Eileen , O'Bryan Moira

Introduction: The production of functional spermatozoa is a highly complex process that requires tight regulation of gene expression. Pre-mRNA splicing is an essential post-transcriptional process. Over 95% of multi-exon human genes undergo alternative splicing to create diverse arrays of mRNAs from a single gene. Disturbance of splicing regulation is associated with many human diseases.Materials and methods: A genome-wide ENU mutagenesis screen was carr...

ra0001p202 | (1) | WCRB2014

Different roads to fertility: distinct molecular and physiological strategies warrant the phenotype ‘high fertility’ in two outbred mouse models

Weitzel Joachim , Sobczak Alexander , Michaelis Marten

Animal models are valuable tools in fertility research. Worldwide >800 transgenic or knockout mouse models are available showing a reproductive phenotype; almost all of them exhibit an infertile or at least subfertile phenotype. In contrast, animal models revealing an improved fertility phenotype are barely described. Here, we present data on two outbred mouse models showing a ‘high fertility’ phenotype. These mouse lines have been generated via selection over a ...

ra0001p203 | (1) | WCRB2014

Abstract withdrawn....

ra0001p204 | (1) | WCRB2014

Recovery of porcine sperm from fetal testicular tissue xenografted into nude mice

Kikuchi Kazuhiro , Noguchi Junko , Kaneko Hiroyuki

Porcine spermatogonia can develop to sperm in neonatal testicular tissue that were cryopreserved and grafted into nude mice. Live piglets were born from zygotes produced by intracytoplasmic sperm injection using these sperm (Kaneko et al. PLoS ONE, 2013). Utilization of fetal tissue will give us valuable chances for conservation of genetic resources and also for improvement of testicular xenografting. We aimed to examine whether porcine fetal testis can produ...

ra0001p205 | (1) | WCRB2014

The impact of oxidative stress on chaperone-mediated human sperm–egg interaction

Grace Bromfield Elizabeth , Nixon Brett , Redgrove Kate , Aitken Robert John

Introduction: Defective sperm–egg recognition is recognised as one of the major causes of failed fertilization in IVF programs. Our recent findings have revealed that such defects may be linked to the molecular chaperone, heat shock protein A2 (HSPA2), due to its ability to facilitate the formation of multimeric zona pellucida (ZP)-receptor complexes on the surface of human spermatozoa. This study aimed to examine the impact of oxidative stress on the formation of these Z...

ra0001p206 | (1) | WCRB2014

Prostaglandin J2 added to artificial insemination sperm dosage reduces the expression of the inflammatory genes in uterus sow

Izquierdo Rico Maria Jose , Ross J W , Soriano-Ubeda C , Hernandez-Caravaca I , Vieira L , Matas C , Garcia-Vazquez F A

Introduction: Billions of sperm are used during porcine artificial insemination (AI) but only a few thousand are able to reach ampullary-isthmic junction of the oviduct to facilitate fertilization. One of the mechanisms which reduce this sperm population is the inflammatory reaction that takes place within the uterus following insemination. Prostaglandin J2, also known as 15-deoxy-Delta-12,14-prostaglandin J2 (15d-PGJ2) is a recently discovered...

ra0001p207 | (1) | WCRB2014

Effect of extracellular long-chain fatty acids on mouse sperm motility

Moriyama Ryutaro , Wakasa Ikumi , Nakagawa Masako

Free fatty acids are not only energy fuels but also signaling molecules. In fact, long-chain fatty acids (LCFAs) have been identified as the ligands for G protein-coupled receptor 40 and 120 (GPR40 and 120). We have previously observed that GPR120 mRNA is expressed in the mouse sperm head. Based on the result, we hypothesized that the LCFAs act as signaling molecules to regulate sperm motility through GPR120. The aim of present study is to clarify whether extracellular LCFAs a...

ra0001p208 | (1) | WCRB2014

Peroxisomes are essential for regular spermatogenesis

Brauns Ann-Kristin , Kamalyan Lilit , Baumgart-Vogt Eveline , Luers Georg H

Peroxisomes are cell organelles with important functions in the metabolism of lipids and reactive oxygen species. In germ cells, they have only recently been described by our groups. Their role for spermatogenesis has not been characterized in detail yet. We have established a mouse model with a conditional knockout of Pex13 in pre-meiotic germ cells to analyse the functions of peroxisomes for development and differentiation of male germ cells. The peroxisomal membran...

ra0001p209 | (1) | WCRB2014

Effect of local treatment of seminal vesiculitis on the quality of equine frozen semen

Sancler-Silva Yame F R , Monteiro Gabriel A , Neto Carlos Ramires , Freitas Dell'aqua Camila P , Crespilho Andre M , Papa Frederico O , Oliveira Pedro V

The seminal vesiculitis leads to contamination of the semen with inflammatory cells, causing agglutination, precipitation, decreased sperm motility and increased reactive oxygen species (ROS). This deleterious effect is most pronounced during the cryopreservation process. Therefore, the aim of this study was to evaluate the quality of frozen semen from stallions with seminal vesiculitis after local treatment. Five stallions with seminal vesiculitis were used and local treatmen...

ra0001p210 | (1) | WCRB2014

Proteomic analysis of human testicular interstitial fluid reflects disordered spermatogenesis in Klinefelter’s syndrome

McLachlan Robert I , Stephens Andrew N , Rainczuk Adam , Foo Caroline , Condina Mark R , Ishikawa Tomomoto , Weidner Wolfgang , Stanton Peter G

Introduction: Primary spermatogenic failure is the commonest cause of male infertility. Changes in Sertoli and germ cell function may be reflected in the testicular interstitial fluid (TIF) proteome and its analysis may provide insights into the pathophysiology. Microdissection testicular sperm extraction (microTESE) provides sperm for ICSI in ~50% of non-obstructive azoospermia (NOA) cases. As the first step in constructing a diagnostic approach for broader application in NOA...

ra0001p211 | (1) | WCRB2014

The role of the molecular chaperone HSPA2 in unexplained failure of IVF

Nixon Brett , Bromfield Elizabeth , Redgrove Kate , Baker Mark A , Aitken Robert John

Introduction: A common defect encountered in the spermatozoa of male infertility patients is an idiopathic failure of sperm–egg recognition. The aim of this study was to investigate the molecular basis of this condition.Materials and methods: For the purpose of this study we compared the proteomic profiles of spermatozoa from male infertility patients exhibiting an impaired capacity for sperm–egg recognition with that of sperm from males of pro...

ra0001p212 | (1) | WCRB2014

Androgen receptor signalling in testicular Leydig cells is essential for Leydig cell maturation and survival

O'Hara Laura , McInnes Kerry , Simitsidellis Ioannis , Morgan Steph , Milne Laura , Mitchell Rod , Smith Lee

Androgen receptor (AR) is expressed by testicular somatic cells including steroidogenic Leydig cells (LC). Its role in LC is ill defined, with hypotheses based on evidence from total androgen receptor knockout models implying that LC AR is responsible for final LC number, regulation of steroidogenic enzyme expression and LC maturation.To better define the role of AR in LC, we generated a mouse line with a LC specific ablation of AR (LCARKO) using the Cre...

ra0001p213 | (1) | WCRB2014

Semen collection and preservation in the salt water crocodile Crocodylus porosus

Johnston Stephen Douglas

Successful semen collection and preservation is vital to the development of a crocodile artificial insemination program for the purpose of intensive production or endangered species breeding. We describe a manual cloacal palpation method for the safe and repeatable recovery of semen from Crocodylus porosus along with corresponding seminal characteristics. Experiments were also conducted to investigate fundamental sperm physiology, including the effect of excessive dil...

ra0001p214 | (1) | WCRB2014

The association between spatial distribution patterns of SPACA1 in human ejaculated sperms and outcomes of conventional IVF

Kishida Kazumi , Harayama Hiroshi , Kimura Fuminori , Murakami Takashi

Introduction: The aims of this study were to show the existence of individual differences in the spatial distribution patterns of sperm acrosome-associated 1 protein (SPACA1) in human ejaculated sperms and to reveal a correlation between distribution pattern of SPACA1 and outcome of conventional IVF.Materials and methods: Informed consent was obtained from all the participants for this study. Viable sperms obtained by swim-up method were selected morphol...

ra0001p215 | (1) | WCRB2014

Detection of potassium channel defects in ICSI patients by patch clamp electrophysiology

Brown Sean G , Mansell Steven , Williams Hannah , da Silva Sarah Martins , Publicover Stephen , Wilson Stuart , Barratt Christopher

Introduction: Potassium channels are essential for the physiological regulation of cell excitability through control of membrane hyperpolarisation. In human sperm slo1 and/or 3 have been proposed to mediate this function (Mannowetz et al. 2013, Mansell et al. 2014). However, to date there is no information on dysfunction in men and the consequence of this for their fertility.Materials and methods: Men undergoing ICSI treatment or recall...

ra0001p216 | (1) | WCRB2014

Spermatogenesis in pubertal common eland (Taurotragus oryx, Pallas 1766)

Pintus Eliana , Ros-Santaella Jose Luis , Kotrba Radim

Introduction: Timing of sexual maturity is essential in order to establish when reproductive technologies can successfully be performed. To date, little is known about reproductive biology in the Taurotragus genus, which includes only two species: common eland (Taurotragus oryx) and giant eland (Taurotragus derbianus). Here, for the first time, we explored testicular development throughout puberty in common eland.Materials and ...

ra0001p217 | (1) | WCRB2014

Evaluation of the binding capacity of fresh and frozen semen to the oviductal cells cultured in vitro

Carneiro Joao Alexandre Matos

The binding of sperm to the oviductal sperm reservoir appears to be an important tool to increase the population responsible for the fertilization of the oocyte cells. However, frozen sperm can lose this function due to the injuries resulting from the freezing process, therefore the aim of this study was to evaluate the binding capacity of fresh and frozen sperm to the oviduct epithelial cells (OEC), cultured in vitro. The semen from ten Andalusian stallions were used...

ra0001p218 | (1) | WCRB2014

Abstract withdrawn....

ra0001p219 | (1) | WCRB2014

Antioxidant vitamins supplementation decreases motility in both fresh and refrigerated ram semen

Parraguez Victor H , Palomino Jaime , Arancibia Massiel , Cofre Eileen , Urquieta Bessie , Raggi Luis , Gonzalez-Bulnes Antonio

Introduction: Spermatozoa death is accelerated by reactive oxygen species in fresh, refrigerated or frozen/thawed semen. In sheep, antioxidant therapy may alleviate the effects of oxidative stress by favoring antioxidant status. Our aim was to compare the effect of antioxidant therapy on sperm motility in ram semen conserved at both ambient (AT) and refrigeration (RT) temperatures.Materials and methods: Five rams were daily supplemented with vitamins C 6...

ra0001p220 | (1) | WCRB2014

Crosstalk of TGFβ superfamily with Smad1 in testis

Chan Kai-Hui , Galuska Sebastian P , Loveland Kate L , Lutz Konrad

The TGFβ superfamily consists of more than 30 ligands, which exert their multiple functions during early embryonic development and in adult tissues. During spermatogenesis, several TGFβ superfamily ligands are present simultaneously to regulate critical stages of development through shared receptors and overlapping transduction molecules. However, the understanding of this promiscuity of signalling pathways remains unfortunately incomplete. Our aim is to study the me...

ra0001p221 | (1) | WCRB2014

Meiotic and post-meiotic germ cells up-regulate blood–testis barrier function

Olcorn Justine Samantha , Rainczuk Katarzyna Eliza , Foo Caroline , Haverfield Jenna T , Stanton Peter G

Introduction: The blood–testis barrier (BTB) remains semi-permeable to tracers of increasing molecular weight in an adult rat model of spermatogenic re-initiation. Complete closure of the BTB occurred when steps 2–7 round spermatids re-appeared in the epithelium, and coincided with the localisation of a new tight junction (TJ) protein claudin-12 (Cldn12) at the BTB. We hypothesise that meiotic and/or post-meiotic germ cells up-regulate Sertoli cell TJs, and aim to de...

ra0001p222 | (1) | WCRB2014

The RNA-binding protein Musashi-2 (MSI2) controls mRNA processing and translational regulation via interactions with SFPQ and PIWIL1 during mammalian spermatogenesis

Sutherland Jessie M , Sobinoff Alexander P , Redgrove Kate , Davidson Tara-Lynne , Siddall Nicole A , Koopman Peter , Hime Gary R , McLaughlin Eileen

Introduction: RNA-binding proteins (RBP) are important catalysts of post-transcriptional gene regulation. The RBP MSI2 directs stem cell function and division through established roles in cell cycle dynamics and cell fate determination. During the complex process of mammalian spermatogenesis, the directed uncoupling of transcription and translation is essential for the correct development of male gametes. We have previously established that dysregulation of MSI2 during germ ce...

ra0001p223 | (1) | WCRB2014

GATA4 serine 105 phosphorylation is required for testicular steroidogenesis in the mouse

Viger Robert , Sab Alain Boulende , Bouchard Marie France , Bergeron Francis , Brousseau Catherine , Tremblay Jacques J , Pilon Nicolas

GATA4 is a transcription factor required for testicular differentiation and for male fertility. In spite of its crucial roles in male reproduction, how the activity of this factor is regulated has yet to be fully understood. The GATA4 protein is phosphorylated on at least two serine residues. This includes GATA4 S105, which has been reported to modulate GATA4 transcriptional activity in cell line studies. Its contribution to endogenous GATA4 activity in the testis, however, ha...

ra0001p224 | (1) | WCRB2014

Maintenance and amplification of testicular tumors by subcutaneous grafting in zebrafish

Kawasaki Toshihiro , Sakai Noriyoshi

Introduction: Testicular tumors not only facilitate analyses of tumorigenesis in testes but also are a good resource for culturing spermatogonial stem cells (SSCs) that allow us to manipulate genetic modification. In zebrafish, a hypertrophied tumorous testis happens to be found occasionally, but no method for maintenance of the individual tumorous testis has prevented systematic analyses of the characters and the practical use. We describe here the method for maintenance of t...

ra0001p225 | (1) | WCRB2014

Effect of aromatase on sperm penetration and embryo cleavage rate during IVF in pig

Oh Jong-Nam , Hwang Jae Yeon , Lee Dong-Kyung , Lee Chang-Kyu

Introduction: Marker genes for evaluating sperm fertility have been developed during a few decades in various species. The CYP19A1/Cyp19a1 has been considered to one of the most common factor related to sperm quality regardless to species. In our previous report, CYP19A1 mRNA quantity was suggested to sperm fertility marker in pig. So, in this study, we examined whether CYP19A1 level and activity are related to sperm fertility to confirm importance of CYP19A1 in porci...

ra0001p226 | (1) | WCRB2014

Effects of different concentration of Escherichia coli on boar sperm quality and field fertility

Sa Soojin , Choi Sunho , Kim Hyunjong , Cho Kyuho , Hong Joonki , Kim Duwan , Kim Younghwa , Park Juncheol

One of the most frequently isolated bacteria in porcine semen was Escherichia coli. Overgrowth by contaminant bacteria such as E. coli has a deleterious effect on semen quality and longevity. The objective of this study was to determine the effects of E. coli isolated from porcine semen on sperm motility, viability, semen pH, and field fertility in sow. Semen samples were prepared using BTS extender that did not contain antibiotics. E. coli ...

ra0001p227 | (1) | WCRB2014

Single-cell level dynamics of mouse spermatogenic stem cells after transplantation

Nakamura Yoshiaki , Yoshida Shosei

In mice, a technique was developed in which transplanted spermatogenic stem cells (SSCs) reconstitute complete spermatogenesis in the testes of infertile recipient. However, the dynamics of SSCs during the reconstitution process after transplantation is largely unknown. To dissect post-transplantation dynamics of SSCs, the behavioral process of donor spermatogonia was observed by in vivo live-imaging, and their fate was analyzed at a single-cell resolution. For live-i...

ra0001p228 | (1) | WCRB2014

Effect of ascorbic acid 2-glucoside supplementation on sperm characteristics during semen cryopreservation in Korean native pig

Park Juncheol , Sa Soojin , Choi Sunho , Kim Hyunjong , Cho Kyuho , Hong Joonki , Kim Duwan , Kim Younghwa

Ascorbic acid 2-glucoside (AA-2G), a stable ascorbate derivative, is a prominent antioxidant characterized by its stability in an oxidative environment. The objective of this study was to evaluate the effect of AA-2G on sperm motility, viability and acrosome integrity during semen cryopreservation in Korean native pig. Semen samples were diluted into a freezing extender (LEY) containing lactose hen-egg yolk with glycerol and Orvus Es Paste (OEP) and loaded in 0.5 ml plastic st...

ra0001p229 | (1) | WCRB2014

Trehalose in glycerol-free freezing extender preserves mitochondria membrane potential of post-thaw boar spermatozoa

Athurupana Rukmali , Funahashi H

Introduction: Movement of spermatozoa and the penetration of zona-pellucida depend on the energy produced in mitochondria. Major steps of cryopreservation can exert stress on sperm mitochondria membrane. Better post-thaw motility, viability and acrosome integrity of spermatozoa was observed when extended in trehalose in a previous study. Objective was to evaluate the effect of trehalose on mitochondria integrity of post-thawed boar spermatozoa.Materials ...

ra0001p230 | (1) | WCRB2014

Abstract withdrawn....

ra0001p231 | (1) | WCRB2014

Knocking-down detrimental seminal protein PDC-109 minimizes cryoinjury to bull spermatozoa

Srivastava N , Srivastava S K , Ghosh S K , Kumar Amit , Andonissamy J , Ponraj Perumal

Introduction: Seminal PDC-109, most abundant protein in bulls, destabilizes sperm membrane by causing efflux of cholesterol. This detrimental effect is minimized to some extent by addition of egg yolk (EY) in extender. Knocking-down of free floating PDC-109 protein by antibodies (Ab) can be another method not yet reported. Therefore, a study was designed to assess the effect of antibodies-mediated sequestration of PDC-109 protein as compared to EY on minimization of cryoinjury...

ra0001p232 | (1) | WCRB2014

Ultrastructural analysis of epididymis of agouti (Dasyprocta spp) during sexual development

Arroyo Maria Angelica M , Santos Paulo Ramos Silva , Oliveira Moacir Franco , de Assis Neto Antonio Chaves

The agouti is a rodent found in the South America and one of the most predated species in Brazil, which has shown a significant decrease of population size. The principal function of epididymis is maturation of spermatozoa, in addition to conducting the same for the vas deferens. The study of sperm pathway, especially of epididymis, can promote morphophysiological information necessary for reproduction biotechnologies. The aim of this study was to describe the ultrastructure o...

ra0001p233 | (1) | WCRB2014

Ultrastructural analysis of spermatozoa in agoutis during sexual development

Arroyo Maria Angelica M , Santos Paulo Ramos Silva , Oliveira Moacir Franco , de Assis Neto Antonio Chaves

The study of the sperm is particularly relevant to reproduction of the species and can promote morphological information necessary for reproduction biotechnologies. The agouti is a rodent found in the South America and represents an excellent source of animal protein. In captivity, the establishment of pubescent occurs at 9 months of age. The aim of this study is to describe the ultrastructure of sperm during sexual development of agoutis kept in captivity. The testes of anima...

ra0001p234 | (1) | WCRB2014

Characterisation of Gankyrin expression in testicular germ cell cancer

Moll Maria Elena Camacho , Macdonald Joni , Mitchell Rod Thomas

Introduction: Testicular germ cell cancer (TGCC) has been increasing in incidence over recent decades. We have previously shown that the precursor lesion for TGCC, carcinoma in situ (CIS) cells, have different phenotypes which determine proliferation rate. CIS which express only OCT4, a well-established marker for CIS, proliferate more that those expressing OCT4 and MAGE-A4 (differentiated germ cells marker). Gankyrin is an oncogene that interacts with MAGE-A4 and sup...

ra0001p235 | (1) | WCRB2014

Tamoxifen inducible Cre systems: adverse effects of low dose tamoxifen treatments on the male reproductive tract

Patel Saloni H , Milne Laura , Sharpe Richard , Smith Lee B

Introduction: Tamoxifen inducible Cre systems have been used to study development specific roles of genes in the testis as they allow tight temporal control of genetic manipulation. However, tamoxifen is an anti-estrogen that competitively binds estrogen receptors. Despite the antagonistic properties of tamoxifen, it also acts as a weak estrogen agonist, hence exerting estrogenic effects in a tissue and cell specific manner. Given the duality of tamoxifen function and the impo...

ra0001p236 | (1) | WCRB2014

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ra0001p237 | (1) | WCRB2014

Involvement of peroxisomes in steroid synthesis in Leydig cells and their role for male fertility

Kamalyan Lilit , Vijayan Vijith , Brauns Ann-Kristin , Colasante Claudia , Schuler Gerhard , Luers Georg H , Baumgart-Vogt Eveline

Introduction: Peroxisomes are ubiquitous organelles, which play an essential role in human physiology. Besides other metabolic pathways, peroxisomes are involved in β-oxidation of fatty acids, ROS-metabolism, and cholesterol synthesis, wherefore alterations of peroxisomal metabolism might affect steroid metabolism and the regulation of corresponding signaling pathways. Indeed male patients with peroxisomal dysfunction exhibit either cryptorchidism or a range of testicular...

ra0001p238 | (1) | WCRB2014

Targeting high mobility group box protein 1 ameliorates experimental autoimmune orchitis

Aslani Ferial , Schuppe Hans-Christian , Guazzone Vanesa A , Bhushan Sudhanshu , Lustig Livia , Meinhardt Andreas , Fijak Monika

High mobility group box protein 1 (HMGB1) plays an important role in onset and progression of autoimmune diseases. This nucleosomal protein is actively secreted during inflammation and acts as a late phase cytokine. Targeting HMGB1 by ethyl pyruvate (EP) significantly reduces its pro-inflammatory effects. Experimental autoimmune orchitis (EAO) serves as a rodent model to study autoimmune based chronic testicular inflammation characterized by elevated levels of pro-inflammatory...

ra0001p239 | (1) | WCRB2014

Effect of addition post-thaw of homologous seminal plasma on quality of donkey frozen semen

de Luna Freire Oliveira Pedro Victor , Oliveira Jose Victor , Neto Carlos Ramires , Sancler-Silva Yame F R , Freitas Dell'aqua Camila P , Papa Frederico O

A recent study showed increase of pregnancy rates in donkey frozen semen with addition of seminal plasma post-thaw. The aim of the present study was to evaluate the effect of addition post-thaw of seminal plasma on donkey frozen semen quality. Five ejaculates from three donkeys were used. Semen was diluted (1:1) with a milk-based extender (Botu-Semen). The semen was frozen with Botu-Cryo extender in isothermal box in straws containing 100×106 of total sperm. Th...

ra0001p240 | (1) | WCRB2014

Difference between of homologous and heterologous seminal plasma addition on post-thawn donkey frozen semen

de Luna Freire Oliveira Pedro Victor , Oliveira Jose Victor , Sancler-Silva Yame F R , Neto Carlos Ramires , Freitas Dell'aqua Camila P , Papa Frederico O

One study showed increased fertilization in IVF when added seminal plasma of bulls with high fertility rates in semen of low fertile. The aim of the present study was to evaluate the difference of addition homologous and heterologous seminal on donkey frozen semen. Five ejaculates from two donkeys were used. Semen was diluted (1:1) with a milk-based extender (Botu-Semen). The semen was frozen with Botu-Cryo extender in isothermal box in straws containing 100×106</sup...

ra0001p241 | (1) | WCRB2014

Evaluation of donkey frozen semen quality in different freezing conditions

de Luna Freire Oliveira Pedro Victor , Oliveira Jose Victor , Sancler-Silva Yame F R , Neto Carlos Ramires , Oliveira Marcelo L F , Papa Frederico O

There are few studies with donkey semen in the literature. In horses, is related that no difference are in freezing semen using automated systems or Styrofoam box. The aim of the present study was evaluate donkey frozen semen quality in different freezing conditions. Five ejaculates from there donkeys were used. Semen was diluted (1:1) with a milk-based extender. The pellet was resuspended with Botucrio and the straws were packaged. The straws were kept in a refrigerator at 5 ...

ra0001p242 | (1) | WCRB2014

Acute reduction of Sertoli cell numbers during development leads to a subsequent reduction in sperm numbers in adulthood

Darbey Annalucia L , O'Shaughnessy Peter , Pitetti Jean-Luc , Nef Serge , Smith Lee , Rebourcet Diane

Sertoli cells (SC) are key initiators of testis development and have a fundamental role in spermatogenesis. However, it is uncertain how an acute reduction in SC number, in a hormone independent-manner at key points in development or adulthood, will impact testis function.To induce acute SC-ablation at key points of testicular development, we used Cre-loxP recombinase technology and the expression of Diphtheria toxin and its receptor. We utilized these l...

ra0001p243 | (1) | WCRB2014

Differential expression of proteins from the human sperm head and its relation to infertility

Baruah Binita Basnet , Kola Srujana , Rukmangadachar Lokesh , Chaturvedi Pradeep , Srinivasan A

Introduction: Male infertility is one of the major challenges in reproduction. The nature of sub-fertility due to male is as complex as that of female and contributes to about 40–50% of infertility cases in human.Objective: We propose to identify all the protein molecules that differ in the different region of the human spermatozoa, i.e. the head and the tail. The aim of the study was to identify the differentially expressed protein in the head frac...

ra0001p244 | (1) | WCRB2014

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ra0001p245 | (1) | WCRB2014

Study of spermatogonia stem cells from domestic animals: derivation and in vitro culture

Song Hyuk , Lee Won Young , Lee Kyung Hoon

Introduction: It is necessary to establish and culture of spermatogonia stem cells (SSCs) in vitro to perform the meiotic differentiation study and to generate transgenic spermatozoa in vitro or ex vivo. This study was focused on in vitro derivation and establishment of SSCs from porcine and canine.Materials and methods: Five day-old porcine and 2-month-old canine testes were used to isolated SSCs and testicular somati...

ra0001p246 | (1) | WCRB2014

Proteomic approach of cryo-damage in bovine spermatozoa

Yoon Sung-Jae , Kwon Woo-Sung , Rahman Saidur , Lee June-Sub , Park Yoo-Jin , You Young-Ah , Pang Myung-Geol

Cryopreservation is the most promising approach for storing spermatozoa. However, it could lead substantial damage to spermatozoa. We present here a couple of functional tests and 2DE PAGE proteome map of bovine spermatozoa in order to evaluate the effect of each cryopreservation procedure, e.g. fresh semen, after cooling, adding cryo-protectant (CP) and thawing on spermatozoa. Our result demonstrated that the cryopreservation procedure reduced the motility (%), viability (%) ...

ra0001p247 | (1) | WCRB2014

Effect of Sertoli cell ablation on testicular function in adult mouse

Rebourcet Diane , O'Shaughnessy Peter , Monteiro Ana , Mitchell Rod , Milne Laura , Smith Lee

The aging of western societies, and the associated increase in obesity, brings with it an increase in prevalence of disorders such as metabolic syndrome which are associated with reduced androgen levels in men. As Leydig cells are the source of androgens in the male, establishing the mechanisms which control Leydig cell development and function is crucial to our understanding of aging and male health. Sertoli cells regulate testicular fate in the differentiating gonad and are ...

ra0001p248 | (1) | WCRB2014

Comparision of DNA fragmentantion and plasma membrane integrity between chilled and frozen semen of bulls

de Mello Papa Patricia , Neto Carlos Ramires , Guasti Priscilla Nascimento , Maziero Rosiara Rosaria Dias , Sancler-Silva Yame F R , Monteiro Gabriel A , Dell'aqua Camila P Freitas , Junqueira Cassio Renesto , Martin Ian , Alvarenga Marco Antonio , Papa Frederico O , Aqua Junior Jose Antonio Dell'

In fixed time artificial insemination (FTAI) is frozen semen is commonly used, however the use of cooled semen is a practice alternative to reduce costs. The aim of this study was compare the difference in DNA fragmentation and plasma membrane integrity between chilled and frozen semen of bulls. Two ejaculates from 20 bulls were collected. Semen was diluted in extender with egg-yolk and glycerol (Botu-Bov™) and divided in two groups: chilled group (CG) samples were cooled...

ra0001p249 | (1) | WCRB2014

Acceptability of New Zealand imported semen to the Japanese dairy farmers

Ozawa Takeyuki , Tamagawa Satomi

Introduction: The USA and the Canadian type of Holstein Friesian semens are widely utilized in Japan. However, imported semens from New Zealand (NZ) have started to utilize among some of Hokkaido dairy farmers gradually. The objective of this study is to clarify reasons and acceptability of NZ semen to the Japanese dairy farmers.Materials and methods: Interviews are held to the NZ semen trading company and nine dairy farmers in Hokkaido, Japan, who are t...

ra0001p250 | (1) | WCRB2014

Exome sequencing identified QRICH2 as a candidate gene responsible for stump-tailed sperm defects in Chinese consanguineous male offspring

Yue Huanxun , Sun Dan , Sun Huaqin , Xu Wenming

Introduction: Dysplasia of the fibrous sheath (DFS) and stump-tail sperm is a rare sperm abnormality often associated with parental consanguinity and found to be associated with recessive mode. However, the molecular pathogenesis remain elusive.Material and methods: Whole-exome sequencing was performed in a typical pedygree with two affected brother. Multiple-tissue real time PCR was used to check the expression of QRICH2 in human multiple tissues. Sperm...

ra0001p251 | (1) | WCRB2014

Manipulating the germ stem cell ‘niche’ to elucidate the origins of testicular germ cell cancer

Macdonald Joni , Mitchell Rod

Germ stem cells in the testis reside in a unique microenvironment, known as the ‘niche’. In humans testicular germ cell cancer (TGCC) is thought to arise when the ‘niche’ fails to induce germ cell maturation. Gonocytes that fail to differentiate into spermatogonia during fetal life form pre-neoplastic carcinoma in-situ (CIS) cells. CIS cells then transform into an invasive tumour in adulthood. Despite the lack of an animal model of TGCC it may be p...

ra0001p252 | (1) | WCRB2014

Roles of testicular macrophage in Leydig cell proliferation in colony-stimulating factor treated mice

Tsai Tina , Sauter Kristin , Milne Laura , Hume David , Smith Lee

Introduction: Leydig cells (LCs), the key producer of testosterone (T) in the testis, are affected by various factors such as hormone stimulation. There are various testicular cells that also affect LC function such as testicular interstitial macrophages (TIMs). TIMs are immune factor-producing cells, located adjacent to the LCs, sharing a close physiological and functional relationship. TIMs are associated with development and function of LCs. Ablation of TIMs have shown to i...

ra0001p253 | (1) | WCRB2014

Panel of MABs to sperm surface proteins as tool for monitoring of sperm–zona pellucida receptors localization and identification

Zigo Michal , Dorosh Andriy , Pohlova Alzbeta , Jonakova Vera , Sulc Miroslav , Postlerova Pavla

Introduction: Primary binding of the sperm to the zona pellucida (ZP), or its analogues, is one of the many steps necessary for successful fertilization in all sexually reproducing species. Sperm bind ZP by means of membrane receptors which recognize carbohydrate moieties on ZP glycoproteins according to a well-precised sequential process. Primary-binding receptors are localized throughout the acrosomal region of the sperm surface of which many have been disclosed in various m...

ra0001p254 | (1) | WCRB2014

Buffalo (Bubalus bubalis) spermatozoa based biosensor for the detection of Cereulide toxin from milk powder

Bamnia Meenakshi , Kaul Gautam

Introduction: Mammalian cell based biosensor consists of biological entity which can be whole mammalian cell, i.e. gametic or somatic cells that interacts with an analyte and produces a signal which can be measured fluorimetrically, colorimetrically, etc. Mammalian cell based biosensor not only helps in the detection but also detect physiological changes Spermatozoa based biosensor rapidly interpret the inherent interaction of spermatozoa with cereulide in a physiologically re...

ra0001p255 | (1) | WCRB2014

Canine spermatogonial stem cells transfected with GFP reporter gene transplanted in recipient teste mice

Pieri Naira Caroline Godoy , de Souza Aline Fernanda , Mancanares Ana Carolina Furlanetto , Casals Juliana Barbosa , Roballo Kelly Cristine , Bressan Fabiana , Oliveira Lilian Jesus , Ambrosio Carlos Eduardo , Martins Daniele Santos

Introduction: Spermatogonial stem cells (SSCs) characterized by ability to self-renew and proliferated, differentiated, and transmitted genetic information. In canine the first attempt (1999) of xenotransplantation into mice did not successfully produce spermatozoa. However, there are evidence that xenogeneic transplant of testis cells can engraft in host testis, and generate donor derived sperm; it suggests the SSCs transplantation may offer a similarity to transgenesis in th...

ra0001p256 | (1) | WCRB2014

Mass-specific metabolic rate influences sperm performance through energy production in mammals

Tourmente Maximiliano , Roldan Eduardo R S

Introduction: Mass-specific metabolic rate, the rate at which an organism consumes energy per gram of body weight, is negatively associated with body size in metazoans. Since the basal metabolic rate of a whole organism represents the sum of its cellular metabolic rates, small species have higher cellular metabolic rates and are capable of processing resources at a faster rate than large species. Recent studies have shown that mass-specific metabolic rate positively influences...

ra0001p257 | (1) | WCRB2014

The role of estrogen and the effect of age on porcine testicular extracellular matrix

Irving-Rodgers Helen , James Chantel , Hummitzsch Katja , Schwartz Jeff , Rodgers Ray , Conley Alan , Berger Trish

Introduction: Extracellular matrix (ECM) has both structural and regulatory roles in tissues and influences the activity of local growth factors. In testis, ECM plays an integral role in the differentiation of germ cells and the function of support cells. It is becoming clear that sex steroids influence ECM and growth factors. In swine, suppression of aromatase activity resulted in an estrogen receptor-mediated increase in Sertoli cells (Berger et al. 2013), with sign...

ra0001p258 | (1) | WCRB2014

Testicular chemical pollutants alter sperm vitality, motility, and morphology: comparative studies in the human and dog

Byers Andrew , Sumner Rebecca , Richardson Holly-May , Stratford Claire , Cammack Thomas , Moxon Rachel , Tomlinson Mathew , Naeem Asad , Craigon Jim , England Gary C W , Lea Richard

Introduction: Declining human and canine male fertility has been associated with exposure to environmental chemicals (ECs). We investigated the effects of ECs on canine and human sperm vitality, motility, and morphology.Methods: Mean canine testicular levels (MTL) of PCB153 and DEHP were 0.9±0.14 μg/kg (n=8) and 0.31±0.03 μg/g (n=5) respectively. Spermatozoa from dogs (n=7) and human donations (n=9,...

ra0001p259 | (1) | WCRB2014

SSP411 a sperm factor involved in oocyte cleavage

Liu Miao , Gu Yihua , Wang Jian , Lu Yi , Yuan Yao , Wu Bin , Chen Ching-Ling C , Shi Huijuan

Introduction: Encoding an 88 kDa protein with a thioredoxin-like domain at the N-terminal region, ssp411 is predominantly expressed at several specific stages from round spermatids to spermatozoa in the rat testis at both mRNA and protein levels. SSP411 is conserved in many species, and can be localized in the testis and mature sperm of human, mouse, and rat.Materials and methods: A line of ssp411 knockout mice (ssp411&#87...

ra0001p260 | (1) | WCRB2014

In vitro effects of hydrogen peroxide on ALF expression in male mouse germ cells

Habas Khaled S A , Anderson Diana , Brinkworth Martin H

Introduction: ALF is a germ cell-specific counterpart of the large (α/β) subunit of general transcription factor TFIIA and has a vital role in the meiotic and post-meiotic development of male germ cells. We have found in vitro a variety of effects of hydrogen peroxide (H2O2) on different types of germ cells, including the induction of DNA strand breaks and apoptosis. We therefore examined its effect on the expression of genes such as AL...

ra0001p261 | (1) | WCRB2014

Time-lapse imaging as a tool to evaluate contractile cell function and sperm transport

Mietens Andrea , Tasch Sabine , Eichner Gerrit , Feuerstacke Caroline , Middendorff Ralf

Introduction: Testicular peritubular cells and the smooth muscle cell layer surrounding the epididymal duct are responsible for the transport of spermatozoa and thus contribute to maintaining male fertility. The intracellular second messenger cGMP mediates smooth muscle cell relaxation and components of cGMP signaling were found in smooth muscle cells of the testis and epididymis. A time-lapse video approach was used to study contractility of seminiferous tubules and the epidi...

ra0001p262 | (1) | WCRB2014

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ra0001p263 | (1) | WCRB2014

Comparison between egg yolk-based and coconut water-based extenders to freeze buffalo semen

Zorzetto Mariana , Sancler-Silva Yame , Dell'aqua Camila Freitas , Neto Carlos Ramires , Zocca Saulo , Ramos Alcides , Oba Eunice , Derussi Ana Augusta Pagnano

The aim of this experiment was compare two egg yolk-based (Tris and botu-Bov) and a coconut water-based (ACP-111) extenders to freeze buffalo semen. Four ejaculates of four buffalo bull were collected. Semen was divided in three groups: EY1 group, the semen was diluted with Tris-Yolk extender; EY2 group, the semen was diluted with Botu-Bov TM; CW group, the semen was diluted with ACP-111TM. The samples were packed in 0.5 ml straws containing 30×10<su...

ra0001p264 | (1) | WCRB2014

Placental phenotype of Turner mouse model: differences between XmO and XpO

Lim Shujing Jane , Ojarikre Andrew , Chuva de Sousa Lopes Susana M

Background: Turner syndrome in human arises from the loss of genetic material from a sex chromosome, resulting in the 45,XO genotype, with the remaining X chromosome being maternally-inherited (Xm) or paternally-inherited (Xp). As the gene dosage and expressions of Xp and Xm chromosomes are epigenetically different, this study aims to investigate the different influences of Xm and Xp on placental phenotype, using a mouse model.Methods: 14 placentas from ...

ra0001p265 | (1) | WCRB2014

The tight junction molecules expression and localization in mouse placenta

Ahn Changhwan , Jeung Eui-Bae

Introduction: Tight junctions (TJs) are composed of a branching network of sealing strands. TJs regulate paracellular conductance and ionic selectivity. The established paracellular transport barriers that control transportation of molecules within intercellular space.Methods: We examined expression of mouse placental claudin family. Pregnant C57/BL6 mice were used in this study and TJ proteins including Claudin-1 to Claudin-24 expressions. The pregnant ...

ra0001p266 | (1) | WCRB2014

The acyl-CoA dehydrogenase, very long chain (ACADVL) expression under hypoxic condition in human placental cell line (BeWo)

Shin Eun-Kyeong , Lee Dongoh , Jeung Eui-Bae

Introduction: Pre-eclampsia (PE) is a medical condition characterized by high blood pressure and significant amounts of protein in the urine of a pregnant woman. Hypoxia can result from a failure at any stage in the delivery of oxygen to cells. In peripheral tissues, oxygen again diffuses down a pressure gradient into cells and their mitochondria, where it is used to produce energy in conjunction with the breakdown of glucose, fats, and some amino acids....

ra0001p267 | (1) | WCRB2014

The regulation of calcium transport channels, TRPV5, TRPV6, PMCA1, NCKX3, and CaBP-9k in ro410960-treated mice of pregnancy

Yang Hyun , Choi Young-Kwon , Jeung Eui-Bae

Introduction: Preeclampsia is a pregnancy-specific disease characterized by concurrent development of hypertension, proteinuria, and oxidative stress in the placenta. Preeclampsia-like genetic models were also developed by modification of preeclampsia-related genes, such as catechol-O-methyltranferase (COMT).Materials and methods: In this study, we induced COMT inhibition in mice during pregnancy to reproduce physiological conditions associated ...

ra0001p268 | (1) | WCRB2014

Expression and regulation of N-myc downregulated gene 2 in mouse uterus during the peri-implantation period

Zhang Xuan , He Yaping , Sun Zhaogui , Wang Jian

Introduction: A sucessful implantation depends on the exquisite coordination between embryo and maternal uterus. N-myc downregulated gene 2 (NDRG2) belongs to the NDRG gene family, a new class of Myc-repressed genes, and possesses tumor suppression function. Given the similarities between embryo implantation and the growth of cancer cell, the present study was undertaken to examine the expression and regulation of NDRG2 during peri-implantation period.Ma...

ra0001p269 | (1) | WCRB2014

The influence of aldrin and dieldrin on the function of bovine uterus and ovary in vitro

Wrobel Michal H , Grzeszczyk Marlena , Mlynarczuk Jaroslaw , Kotwica Jan

Introduction: Aldrin and its derivative dieldrin belong to the group of chloroorganic pesticides, which due to their lipophilic properties and resistance to biodegradation are accumulated in tissues of animals and humans, and they can mimic the action of endogenous hormones.Materials and methods: Myometrial strips and endometrial, myometrial, granulosa, and luteal cells obtained from cows on days 8–12 of the estrous cycle, were incubated with aldrin...

ra0001p270 | (1) | WCRB2014

PGRMC1 protein expression in the canine endometrium

Bartel Cordula , Walter Ingrid

Introduction: The sex steroid hormone progesterone (P) is the most important hormone in the canine estrous cycle for initiation and preservation of pregnancy. The responsiveness of the endometrial cells to P depends on progesterone receptors (PGRs). In recent studies alternatives to the classical progesterone receptor were identified as membrane-associated progesterone receptors (MAPGRs). Progesterone receptor membrane component-1 (PGRMC-1) is a representative of the MAPGRs wh...

ra0001p271 | (1) | WCRB2014

Do maternal obesity and mood disorders correlate with changes in placental glucocorticoid transporter gene expression?

Mina Theresia H , Davidson Callam , Taylor Ashley , Norman Jane E , Reynolds Rebecca M

Introduction: Both obesity and mood disorders associate with altered glucocorticoid (GC) secretion and action. During pregnancy placenta regulates foetal exposure to maternal GC through enzymatic clearance (HSD-2) and retrograde transfer (ABC transporters). We and others have shown that maternal obesity and/or mood disorders alter placental HSD-2 mRNA level. We therefore hypothesised that placental ABC transporter gene expression is also altered, and investigated this using sa...

ra0001p273 | (1) | WCRB2014

Estradiol promotes differentiation of uterine natural killer cells during establishment of pregnancy

Gibson Douglas A , Critchley Hilary O D , Saunders Philippa T K

Background: Uterine natural killer (uNK) cells are a phenotypically distinct population of tissue-resident immune cells that are reported to regulate vascular remodelling within the endometrium. The mechanisms that control accumulation of uNK cells in the endometrium remain unresolved although there is evidence to suggest that homing and differentiation of peripheral blood NK (pbNK) cells may contribute to the rapid increase in cell numbers during the secretory phase of the me...

ra0001p274 | (1) | WCRB2014

A role for androgens in the regulation of endometrial breakdown and repair in a mouse model of menstruation

Cousins Fiona L , Murray Alison A , Critchley Hilary O D , Saunders Philippa T K

Background: The human endometrium is a dynamic, multi-cellular sex steroid-dependent tissue subject to cyclical episodes of breakdown (menses), repair and regeneration. The mechanisms responsible for initiation of menses are well characterised; the molecular and cellular changes responsible for rapid repair are poorly understood. Re-epithelialisation of the tissue is thought to involve contributions from both the stromal and epithelial compartments. A role for androgens in end...

ra0001p275 | (1) | WCRB2014

Apoptosis and neutrophil infiltration in the human endometrium at menstruation, recapitulated in mouse model of induced menstruation

Armstrong Gregory M , Murray Alison , Maybin Jacqueline A , Rossi Adriano G , Critchley Hilary O D

Introduction: Menstruation is an inflammatory process characterised by tissue breakdown, bleeding and recruitment of leukocytes. Resolution of this inflammation at menses is critical both to limiting tissue damage and to efficient repair. Apoptosis and clearance of apoptotic cells are necessary for this process, and are thought to involve neutrophils.Our aims were to (i) delineate neutrophil numbers and identify apoptosis in peri-menstrual endometrium an...

ra0001p276 | (1) | WCRB2014

Glucocorticoids trigger macrophage migration inhibitory factor (MIF) secretion by decidualized human endometrial stromal cells in vitro: the modulatory effect of Bisphenol A

Mannelli Chiara , Szostek Anna , Ciaffarafa Sara , Carotenuto Claudiopietro , Ietta Francesca , Romagnoli Roberta , Piotrowska-Tomala Katarzyna , Paulesu Luana , Skarzynski Dariusz

Introduction: Pro-inflammatory cytokines are important mediators of the early phases of gestation. Among all these cytokines, the macrophage migration inhibitory factor (MIF) plays significant roles in pregnancy establishment. In many cell types, low concentrations of glucocorticoids (GCs) are known to enhance MIF secretion, but the effect of GCs on endometrial cells has not been elucidated. Bisphenol A (BPA) is an environmental chemical that can alter reproductive physiology ...

ra0001p277 | (1) | WCRB2014

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ra0001p278 | (1) | WCRB2014

Bovine placental lactogen is cleaved by matrix metalloproteinases and resulted 25k N-terminal fragments inhibit the proliferation of vascular endothelial cells

Sasaki Shobu , Hayashi Ken-Go , Hosoe Misa , Kizaki Keiichiro , Hashizume Kazuyoshi , Takahashi Toru

Bovine placental lactogen (bPL) is a classical member of a prolactin (PRL) gene family expressed in the placenta. bPL exerts lactogenic activity similar to pituitary PRL, however, detailed information about the role of bPL during bovine gestation is still limited. The 16 k N-terminal fragments of PRL, generated by enzymatic cleavage, have angiostatic activities in human and rodents. In the present study, we examined the feasibility of N-terminal fragments of bPL following clea...

ra0001p279 | (1) | WCRB2014

An estradiol metabolite 2-methoxyestradiol exhibits estrogen-like effect in vitro and in vivo

Lee Ji-Sun , Jeung Eui-Bae

Introduction: 2-Methoxyestradiol (2-ME) is an endogenous metabolite of 17-estradiol (E2) that interacts with estrogen receptors (ERs) and microtubules, and it has a low affinity for ERs. It has been identified as a potential novel antitumor agent combining anti-proliferative activity on a wide range of tumor cell types with anti-angiogenic actions. Also, 2-ME attracted considerable interest as a potential anti-cancer therapeutic.Materials and ...

ra0001p280 | (1) | WCRB2014

Prominin-1 and -2 in uterine epithelial cells during early pregnancy

Dowland Samson , Green Charmaine J , Lindsay Laura A , Day Margot L , Murphy Chris

Introduction: In preparation for blastocyst implantation, the microvilli of uterine epithelial cells (UECs) are lost and the apical plasma membrane flattens. Prominin-1 is a membrane glycoprotein restricted to the apical domain of epithelial cells, while the structurally related prominin-2 is distributed throughout the plasma membrane of epithelial cells in a non-polarised way. This study aimed to examine the expression of prominin-1 and -2 in UECs to examine their potential r...

ra0001p281 | (1) | WCRB2014

Trophoblast-decidual crosstalk optimises trophoblast invasion

Menkhorst Ellen , Gamage Thilini , Van Sinderen Michelle , Cuman Carly , Rainczuk Adam , Stephens Andrew N , Dimitriadis Eva

Introduction: During the establishment of pregnancy, extravillous trophoblast (EVT) must invade through the decidua to form a functional placenta. Impaired decidualization is associated with placental insufficiency, including preeclampsia. The mechanisms by which decidual cells interact with EVT remain largely unknown. We hypothesised that decidual–EVT interactions are critical for appropriate invasion.Methods: Primary human endometrial stromal cell...

ra0001p282 | (1) | WCRB2014

Caveolins redistribute in uterine epithelial cells during early pregnancy: role in epithelial polarisation

Madawala Romanthi J , Dowland Samson , Poon Connie E , Murphy Christopher

Introduction: Uterine epithelial cells (UECs) undergo a number of changes to become receptive to blastocyst implantation including the loss of integrin based focal adhesions (FA), and increase in basolateral plasma membrane (PM) curvature; despite these changes UECs maintain a polarised state. Caveolae are lipid rich invaginations of the PM which contribute to membrane curvature. Major caveolae proteins (caveolin 1, and 2, PTRF, and SDPR) were investigated to explore the possi...

ra0001p283 | (1) | WCRB2014

Interleukin-11 is critical for placentation in mice

Winship Amy , Menkhorst Ellen , Van Sinderen Michelle , Dimitriadis Eva

Introduction: Placentation involves trophoblast cell invasion into the decidua to remodel maternal arteries. Interleukin (IL)-11 is critical for human trophoblast cell migration/invasion in vitro, however its role in placentation in vivo has not been investigated. We hypothesised that IL11 plays a critical role in trophoblast function during placentation. The aim of this study was to examine the effect of IL11 inhibition on placentation in mice.<p class="...

ra0001p284 | (1) | WCRB2014

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ra0001p285 | (1) | WCRB2014

Distinct sex-specific gene expression changes in the human placenta in association with childhood allergy at 2 years

Tuck Astrud , Osei-Kumah Annette , Saif Zarqa , Clifton Vicki L

The prevalence of childhood allergic disease has increased dramatically in developed countries. Modern environmental changes are hypothesised to be causing deviations in fetal programming, in which the placenta plays a central role, increasing the prevalence of disease. We hypothesised that susceptibility to childhood allergy is determined by changes in placental function that programs fetal immune function. In this study we aimed to identify candidate genes and pathways in hu...

ra0001p286 | (1) | WCRB2014

The EpCAM-claudin 7 protein complex decreases during early pregnancy in the rat

Poon Connie E , Day Margot L , Murphy Christopher

Introduction: During implantation, uterine luminal epithelial cells (UECs) disassemble cell–cell and cell–matrix adhesions that contribute to the formation of an epithelial barrier, which is refractory to blastocyst implantation. These changes are critical for successful implantation. We have shown previously that claudin 7 is present in UECs and is lost prior to implantation in the rat. Epithelial cell adhesion molecule (EpCAM) is another protein frequently associat...

ra0001p287 | (1) | WCRB2014

Short-term administration of ulipristal acetate modulates endometrial sex steroid receptor expression and cell proliferation markers

Murray Alison A , Shaw Grace G , Nicol Moira , Williams Alistair R W , Saunders Philippa T K , Critchley Hilary O D

Introduction: Uterine fibroids cause pain, pressure symptoms and heavy menstrual bleeding (HMB). Fibroid growth is sex steroid dependent. The selective progesterone receptor (PR) modulator (SPRM), ulipristal acetate (UPA) reduces fibroid size and alleviates HMB in 90% of women. UPA has both agonist and antagonist properties and induces (S)PRM-associated endometrial changes (PAEC). The mechanism of UPA action and aetiology of PAEC are unknown. We propose that modulation of ster...

ra0001p288 | (1) | WCRB2014

Periconceptional alcohol exposure programs sex specific hyperinsulinemia possibly through dysregulation of placental O-linked glycosylation

Gardebjer Emelie M , McMahon Kathryn , Moritz Karen M , Pantaleon Marie

Introduction: Exposure to an adverse environment around the time of conception can program adult onset disease although mechanisms involved are unclear. Recently we showed that periconceptional alcohol (PC:EtOH) consumption causes placental stress demonstrated by elevated levels of 11βHSD-2, and sex-specific increases in placental glucose transporters and glycogen accumulation. O-linked-N-acetylglucosamine transferase (OGT), an X-linked gene product, res...

ra0001p289 | (1) | WCRB2014

Analysis of EMG signal linear synchronization in porcine uterus

Domino Malgorzata , Gajewski Zdzislaw , Pawlinski Bartosz , Zabielski Romuald , Aniolek Olga

Introduction: Understanding the direction and quantity of information flowing in a complex system of myometrium is a fundamental task in signal processing. The new perception of such related functioning systems can be given by the evaluation of synchronization between signals.Materials and methods: Spontaneous uterine activity was recorded in ten mature Polish Landrace sows directly by commercial implants TL10M3-D70-EEE (DSI, USA) surgically positioned b...

ra0001p290 | (1) | WCRB2014

Sex specific effects of maternal dietary protein upon uterine blood flow and fetal growth

Hernandez-Medrano Juan H , Copping Katrina J , Hoare A S , Wapenaar Wendela , Grivell Rosalie , Callaghan Matthew , McMillen Caroline , Rodgers Ray , Perry Viv E A

Introduction: Neonatal and fetal mortality is increased in male fetuses. Evidence is emerging of sex specific differences in fetal and placental perfusion1. This study investigates effects of dietary intervention at specific time points upon blood flow and fetal growth trajectory in cattle.Materials and methods: Primiparous heifers were either individually- (Australia, Au; n=360) or group-fed (UK; n=188) diets containing diff...

ra0001p291 | (1) | WCRB2014

Effect of pentraxin 3 on inflammation-induced fetal loss

Bonfiglio Rita , Camaioni Antonella , Palone Francesca , Bottazzi Barbara , Sironi Marina , Garlanda Cecilia , Salvatori Giovanni , Mantovani Alberto , Salustri Antonietta

Pentraxin 3 (PTX3) is an essential component of the innate immunity with a non-redundant role in conferring resistance to pathogens and in modulating inflammatory reactions in several diseases. Pregnancy is characterized by mild systemic immunosuppression and inflammation. An uncontrolled immune activation at the fetal–maternal interface caused by bacterial infections or inflammatory stimuli is associated with high risk of miscarriage both in mice and in women. In this st...

ra0001p292 | (1) | WCRB2014

Glycoforms of α-2-macroglobulin in uterine fluid, are altered during the menstrual cycle, and differ between fertile and infertile women

Edgell Tracey A. , Fitzgerald Harriet , Salamonsen Lois

Introduction: Our previous 2D-DIGE studies of uterine fluid, identified differing isoform distributions of the serine protease inhibitor α-2-macroglobulin (A2M) resulting from post-translational modification (PTM’s), among fertile and infertile women. Carbohydrate side chains are a key PTM, often conferring recognition between protein partners, e.g. enzyme and inhibitor. Altered PTM’s may potentially change the interactions of A2M with proteases, effectively alt...

ra0001p293 | (1) | WCRB2014

Non-cytopathic BVDV suppressed innate immune response in bovine endometrial cells exposed to bacterial lipopolysaccharide

Oguejiofor Chike Fidelis , Cheng Zhangrui , Anstaett Olivia , Brownlie Joe , Fouladi-Nashta Ali A. , Wathes D Claire

Introduction: Infection with non-cytopathic BVDV (ncpBVDV) is associated with uterine disease and infertility in dairy cows although the underlying mechanisms are poorly understood. This project investigated the influence of ncpBVDV on innate immunity of the bovine endometrium.Materials and methods: Primary bovine endometrial cells from each of six cows were divided into four groups (Control; LPS; BVDV; BVDV LPS). Cells were infected with ncpBVDV for 4 d...

ra0001p294 | (1) | WCRB2014

Remodelling-associated processes during postpartum uterine involution in mice

Menzies Fiona M , Burton Laura , Ahmed Humera , Oldham Rachel S. , Higgins Claire A , Nelson Scott M , Nibbs Robert J B

Introduction: Following labour, the uterus rapidly returns to its pre-pregnancy state. Exact mechanisms underlying labour and postpartum involution are not fully understood but inflammation is understood to play a key role. It has been hypothesised that rather than initiating or propagating labour, inflammation may prime the uterus for extensive postpartum tissue repair and remodelling, in a manner akin to repair in exercising skeletal muscle. This study aims...

ra0001p295 | (1) | WCRB2014

Investigating the role of androgens in endometriosis

Simitsidellis Ioannis , Greaves Erin , Collins Frances , Horne Andrew W , Saunders Philippa T K

Introduction: Androgen receptors (AR) have been detected in the human endometrium throughout the menstrual cycle and are reported to regulate expression of genes associated with cellular proliferation and cell-cycle regulation. Endometriosis is defined by establishment and growth of endometrial tissue in ectopic locations (typically on the peritoneal wall); patients often experience pain and infertility. In the current study we have used human tissue and a new mouse model of e...

ra0001p296 | (1) | WCRB2014

Evaluation of protein profile in uterine secretion on early cyclic and gestational diestrus in bitches

Derussi Ana Augusta Pagnano , Volpato Rodrigo , de Freitas Guaitolini Carlos Renato , Ackermann Camila Louise , Taffarel Marilda Onghero , Cardoso Guilherme Schiess , Villaverde Ana Izabel Silva Balbin , de Souza Fabiana Ferreira , Lopes Maria Denise

Introduction: During peri-implantation period the canines conceptus are nourished by secretions in uterine lumen, a phenomenon known as histotrophic nutrition, in which several proteins are present. The aim of this study was to compare the protein profile in uterine secretion in early pregnancy and early diestrus cyclic using two-dimensional electrophoresis technique.Material and methods: For that, ten bitches were submitted to OHE on the 12th day after ...

ra0001p297 | (1) | WCRB2014

Does decidual macrophage polarisation affect trophoblast behaviour in co-culture studies?

Buckley Rebecca , Dumitriu Ingrid , Whitley Guy S , Cartwright Judith

Introduction: Decidual macrophages constitute the second largest leukocyte cell type in the decidua during early pregnancy, and have been implicated in the regulation of extravillous trophoblast (EVT) invasion. Macrophage cells in the decidua are thought to have an anti-inflammatory M2 phenotype and the presence of macrophages with a more pro-inflammatory M1 phenotype has been associated with preeclampsia. The aim of this study was to determine whether decidual macrophage pola...

ra0001p298 | (1) | WCRB2014

Decidual stromal cell regulation of trophoblast in first trimester pregnancies with normal and impaired spiral artery remodelling

James-Allan Laura , Wallace Alison E , Whitley Guy S , Cartwright Judith

Introduction: Decidualisation of the endometrium is the differentiation of uterine stromal cells into specialised secretory decidual stromal cells (DSC) primed for pregnancy. If pregnancy occurs, DSC control and regulate fetal trophoblast invasion. Pre-eclampsia is a disorder associated with inadequate trophoblast invasion and spiral artery (SA) remodelling. Uterine artery Doppler resistance index (RI) in the first trimester of pregnancy measures the extent of SA remodelling; ...

ra0001p299 | (1) | WCRB2014

Effect of maternal diet on the amino acid composition of human uterine fluid

Kermack Alexandra Jayne , Cheong Ying , Brook Nick , Macklon Nick , Houghton Franchesca D

Introduction: Preimplantation embryos depend on nutrients provided in the reproductive tract for development. Early human embryos have an increasing requirement for amino acids as they develop from the 1-cell through to the blastocyst stage. In vivo, the uterus provides the final environment for preimplantation development, but surprisingly, the nutritional composition of human uterine fluid has been little investigated. This study aims to examine the amino acid compo...

ra0001p300 | (1) | WCRB2014

Reproductive performance, oxidative enzyme and gonadotropins status of rabbit does fed organic selenium as dietary supplement

Omeje Valentine , Machebe Ndubuisi Samuel , Ezekwe Arinze

Introduction: Selenium deficiency is known to affect negatively the reproductive performance of farm animals including rabbits. This study evaluated the effect of feeding organic selenium (OSe) as dietary supplement on the reproductive performance, oxidative enzyme and gonadotropins status of rabbits does.Materials and methods: Sixteen female rabbit does of 6 to 7 months of age and weighing between 1.95 to 2.03 kg were assigned to four treatment groups r...

ra0001p301 | (1) | WCRB2014

Secreted phosphoprotein 1 (SPP1), vascular endothelial growth factor A (VEGFA), heparinase (HPSE), hypoxia-inducible factor (HIF1A) and uteroferrin (ACP5) gene expression in uterine and placental tissues from Large White-Landrace (LW-LD), hyperprolific Large White (hLW) and Meishan (MS) pigs

Hernandez Silvia C , Hogg Charis O , Ait-Ali Tahar , Ashworth Cheryl Joy

Introduction: This study compared uterine and placental expression of genes implicated in placental attachment and angiogenesis in two prolific (hLW and MS) and one control (LW-LD) pig breeds.Methods: Placental and endometrial tissues supplying the smallest and a normal-sized foetus within the same litter were collected from five LW-LD, five hLW and four MS gilts on days 40–45 of pregnancy. RNA was isolated and qPCR for SPP1, VEGFA...

ra0001p302 | (1) | WCRB2014

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ra0001p303 | (1) | WCRB2014

Effect of progesterone supplementation on endometrial receptivity for implantation in the pig

Blitek Agnieszka , Szymanska Magdalena

Progesterone (P4) produced by the corpus luteum (CL) is required for the attainment of endometrial receptivity and further conceptus implantation. Induction of puberty with PMSG/hCG in gilts results in lower P4 concentration in blood serum, that is accompanied by decreased endometrial expression of genes involved in conceptus-maternal interactions. Therefore, the present study aimed to determine the effect of P4 supplementation in pregnant gilt...

ra0001p304 | (1) | WCRB2014

The relaxing effect of dihydrotestosterone on myometrial smooth muscle contraction is mediated via blockage of intracellular calcium increase and inhibition of myosin light chain phosphorylation

Makieva Sofia , Saunders Philippa T K , Norman Jane E

Background: The impact of androgens on myometrial contraction has received less scrutiny, compared to progesterone and estrogens. We sought to i) examine the effect of dihydrotestosterone (DHT; a non-aromatizable androgen) on stretch-induced contraction of human myometrium and ii) gain mechanistic insights by studying changes in intracellular calcium [Ca2+] and phosphorylation of myosin light chain (MLC).Methods: Myometrial strips were recover...

ra0001p305 | (1) | WCRB2014

Detection of circulating miRNAs in the maternal serum during early pregnancy in the pig by qRT-PCR and digital RT-PCR

P Reliszko Zaneta , Gajewski Zdzislaw , M Kaczmarek Monika

Exosomes are recognized as new mediators of intercellular cell signaling between neighboring cells and distant tissues, acting independently but synergistically with growth factors and hormones. Trophoblast and stromal cells of the villi were identified as a source of exosomal miRNAs released into the maternal circulation. In this study we hypothesized that miRNAs participating in the embryo–maternal dialog are released into the maternal blood stream during early pregnanc...

ra0001p306 | (1) | WCRB2014

Inhibition of the polyamine pathway induces an embryonic diapause-like state in the mouse embryo

Catherine Fenelon Jane , Banerjee Arnab , Murphy Bruce D

Introduction: Embryonic diapause is a period of developmental arrest in which the embryo is maintained in a dormant state for an extended period of time. Over 130 species of mammals undergo embryonic diapause, but the molecular control mechanisms remain unknown. In this study, we investigated the potential role of polyamines during embryonic diapause in the mouse. Polyamines play essential roles in cell growth and proliferation, and the regulation of their expression is contro...

ra0001p307 | (1) | WCRB2014

Effect of flax seed (Linum usitatissimum) extract on modulation of prostaglandin E2 production by buffalo endometrial stromal cells cultured in vitro

Kumar Singh Sanjay , Sharma Chethan G , Nongsiej Jessiehun , Singh R P , Kumar Agarwal Sudhir

Different hormones have been tried to increase embryonic survivability with inconsistent results. Plant based treatment plays positive role in enhancing reproductive functions, acting as precursors for different signalling molecules and influencing cell processes in humans as well as animals, but lacks scientific validation. Effect of aqueous and ethanolic extracts of flax seed on PGE2 modulation by buffalo endometrial stromal cells was studied. Buffalo stromal cell...

ra0001p308 | (1) | WCRB2014

Detection of SPAM1 in the bovine oviductal fluid

Salvador Acuna Omar , Jimenez-Movilla Maria , Vilella Ismael , Jara Lourdes , Canovas Sebastian , Coy Pilar , Aviles Manuel , Izquierdo Rico Maria Jose

Introduction: The sperm adhesion molecule 1 (SPAM1) is a glycoprotein present in the sperm membrane and it is involved in degradation of cumulus matrix and ZP binding. In the mouse, SPAM1 is secreted by the oviduct and the uterus and is acquired by the sperm during its transit through the genital tract. Our aim was to determine the presence of SPAM1 in the bovine oviduct.Materials and methods: Ovaries and oviducts were obtained from slaughterhouse cows a...

ra0001p309 | (1) | WCRB2014

Comparison between the effect of hCG and GnRH analogue (buserelin) treatment post laparoscopic insemination on reproductive performance of ewes

Swelum Ayman Abdel-Aziz , Al-Owaimer Abdullah N , Abouheif Mohamed A

The present study aimed to compare between the effect of hCG and GnRH analogue treatment post laparoscopic insemination on reproductive performance of ewes. Oestrus synchronization was performed for 120 ewes (Najdi=60 and Naeimi=60) using intra-vaginal progesterone sponge and equine chorionic gonadotrophin (eCG). Semen was collected from six proven fertile rams by artificial vagina. Ewes were laparoscopic inseminated 48–50 h after sponge withdrawal,and divided randomly in...

ra0001p310 | (1) | WCRB2014

Novel sequence variations in exon 25 of JHDM2A among overweight and obese sub/infertile males in South India

Najafi Mohsen , Malini Suttur S

Objectives: To analyze three exons of JHDM2A gene in obese males associated with sub/infertility and to compare with non-obese infertile and fertile males as controls in Karnataka, South India.Materials and methods: For the candidate gene analysis, 200 cases with obesity/ and infertility and 50 controls (non-obese fertile males) were selected. After 3–5 days of ejaculatory abstinence the semen samples were collected in a sterile plastic con...

ra0001p311 | (1) | WCRB2014

Incerement of proinflammatory cytokine (IL6) and lipid peroxidation in obese infertile males may lead to more impaired seminogram compared to non-obese infertile males

Najafi Mohsen , Aarabi Mahmoud , Yousefi Azin , Nasri Soroush , Ajami Sanaz , Malini Suttur S

Objectives: The present study measures the level of interleukin-6 (IL6) and the lipid peroxidation (LPO) marker, malondialdehyde (MDA), in the semen ofobese and non-obese infertile males vs obese and healthy fertile males and to correlate their level with the semen characteristics.Materials and methods: A total of 100 infertile participants, consisting of equal number of obese infertile and non-obese infertile males (n=50) were selectedas cases....

ra0001p312 | (1) | WCRB2014

Metallothionein gene expression in green mussels (Perna viridis) as a bioindicator for heavy metal on the North Coast of Java and its impact on the development of reproductive organs

Irnidayanti Yulia , Septiana Shofy , Layli Fitri Apriliana

Jakarta Bay with 13 rivers that flow into, have experienced contamination and is the most polluted bays in Asia as a result of industrial and household waste. At this time, there are approximately five million kinds of chemicals that have been identified and have been known to 60 000 types of them have been used, even 1000 of other kinds and new chemicals freely traded every year. One of the B3 waste is heavy metals. Various ways have been conducted to measure the effects of e...

ra0001p313 | (1) | WCRB2014

Allopregnanolone promotes angiogenesis and inhibits apoptosis in the corpora lutea in rat ovarian cycle

Laconi Myriam Raquel , Cabrera Ricardo Jorge , Parborell Fernanda

We report the effect of allopregnanolone (ALLO, 3α-hydroxy-5α-pregnan-20-one), on morphological changes and on angiogenesis in the rat ovary. ALLO, one of the best characterized neurosteroids, has effects on the reproductive female biology. First, we previously demonstrated that ALLO inhibited ovulation, secretion of LH and progesterone, increased the prolactin levels, inhibited the female receptivity and apoptosis in rat corpus luteum (CL). Now, we show the effect o...

ra0001p314 | (1) | WCRB2014

Abstract withdrawn....

ra0001p315 | (1) | WCRB2014

Protein hormone proteolysis in target cell endosomes and lysosomes and release of previously unrecognized signaling information

Campbell Kenneth L , Haspel Nurit , Dominas Barbara , Mason Briana , Calixte Wagner , Bigos Ronald , Taguzem G Florent , Vader Umaben , Palmstrom Noelle , Steinbruck Jeremy , Solchenberger Sherry , Diallo Fatoumata

Organisms use protein hormones even when smaller molecules can trigger identical transduction paths. Why? Could residual peptides from hormone proteolysis extend the mechanisms of action of these hormones? Protein hormones are endocytosed by target cells and digested by cathepsins (A, B, C, D, F, H, L, O, X) acting sequentially as endosomes/lysosomes move centripetally. In silico prediction of multiple cathepsin action on each of 92 hormones usually leaves 3–24 r...

ra0001p316 | (1) | WCRB2014

Deficiency of monoclonal nonspecific suppressor factor beta (MNSFβ) gene leaded to the early loss of pregancy in mice

He Yaping , Zhang Xuan , Wang Jian , Sun Zhaogui

Introduction: Immune tolerance at the feto–maternal interface must be established during the processes of implantation and pregnancy. Monoclonal nonspecific suppressor factor beta (MNSFβ) is a secreted protein that possesses antigen-nonspecific immune suppressive function. The objective of this study was to establish the MNSFβ gene knockout mouse model, observe and analyze its reproductive phenotypes to further explore the roles of MNSFβ played in mouse emb...

ra0001p317 | (1) | WCRB2014

Analysis of marsupial ZP: molecular and phylogenetic approach

Nicolas C Moros , Rico M J Izquierdo , Holt W V , Chevret P , Lopez-Bejar M , Martinez-Nevado E , Ballesta J , Aviles M

Introduction: The zona pellucida (ZP) is an extracellular matrix that surrounds mammalian oocytes. Marsupial ZP has been considered formed by three proteins. However, the recent description of four glycoproteins (ZP1, ZP2, ZP3, and ZP4) in the ZP of some species of eutherian (rat, human, hamster, rabbit) suggests the need for a reanalysis of the ZP composition in marsupials.Material and methods: Complementary DNA was obtained from ovaries from one Austra...

ra0001p318 | (1) | WCRB2014

Transmission of lethal phenotype in a Mendelian fashion by genetically modified pigs that underwent blastocyst complementation

Matsunari Hitomi , Watanabe Masahito , Umeyama Kazuhiro , Nakano Kazuaki , Nagaya Masaki , Nakauchi Hiromitsu , Nagashima Hiroshi

In our previous study, we demonstrated that pigs expressing neonatally lethal phenotype induced by genetic modification could be rescued by induction of chimerism by using blastocyst complementation. Here, we show that the obtained chimeric pigs can faithfully transmit the genotype, which is the cause of the lethal traits, to progenies.Cloned embryos (male) of pigs showing the phenotype of pancreatogenesis deficiency by expression of the Pdx1-Hes1</e...

ra0001p319 | (1) | WCRB2014

Reproductive and toxic effects of methanol extract of Alchornea cordifolia leaves

Ajibade Temitayo Olabisi

Alchornea cordifolia is commonly used as a medicinal plant throughout its area of distribution in Africa including Nigeria and Democratic Republic of Congo. The leaves are traditionally used for the treatment of veneral diseases. Using 36 male albino rats, randomly divided into six groups of six rats each, the effects of oral administration of the methanol extract of the leaves of A. cordifolia on some reproductive and haematological parameters of male rats w...

ra0001p320 | (1) | WCRB2014

Pinacidil, a KATP channel opening drug, protects human oocytes against metabolic stress

Fernandes Goncalo M T , Gall Mandy , Drew Ellen , Barratt Evelyn , Madamidola Oladipo , Brown Sean G , Milne Alison M , da Silva Sarah Martins , Barratt Christopher , Jovanovic Aleksandar

Introduction: The primary treatment of infertility is assisted reproductive technology (ART). However, removal of an oocyte from its natural environment and exposure to in vitro conditions diminishes oocyte quality during the process of ART. In our preliminary studies we have shown that oocytes express cytoprotective ATP-sensitive K+ (KATP) channels. Therefore, the aim of our study was to elucidate whether activation of these channels protects hu...

ra0001p321 | (1) | WCRB2014

Fetal overexposure to androgens, not estrogens, leaves a permanent legacy in terms of adrenal function in offspring

Ramaswamy Seshadri , Grace Cathal , Howie Forbes , Connolly Fiona , Konovalova Julia , McNeilly Alan S , Duncan W Colin , Rae Mick

Introduction: Overexposure to androgens during fetal life creates offspring with a polycystic ovary syndrome (PCOS)-like phenotype. Whether this phenotype is attributable to androgens during fetal life, or estrogens via maternal/placental metabolism, remains uncertain. The potential for in utero steroidal disruption of adrenal development and function is also unclear. We have examined the adrenal during development and adulthood in response to in utero androg...

ra0001p322 | (1) | WCRB2014

Inhibition of the TLR4 signalling pathway protects from intrauterine heat-killed Escherichia coli-induced pre-term delivery

Chin Peck Yin , Dorian Camilla , Hutchinson Mark , Robertson Sarah

The pathophysiology of preterm labour is poorly understood and the causal factors uncertain, but inflammatory mechanisms are clearly implicated. Escherichia coli contain lipopolysaccharide, a pathogen-associated molecular pattern (PAMP) which binds Toll-like receptor 4 (TLR4) and related TLR-associated receptors to activate inflammation and induce preterm delivery (PTD). This project seeks to investigate whether inhibition of the TLR4 signalling pathway using small mo...

ra0001p323 | (1) | WCRB2014

Effect of doses and time post-injection on hormonal induction of spermiation in Bufo calamita (Anura: Bufonidae)

Lopez Elia Alonso , Kouba Andrew J , Arregui Lucia

Introduction: Currently, one-third of amphibians species are threatened with extinction and assisted reproductive technologies (ART) play an important role in their genetic management. The first step in developing ART is to create protocols for obtaining gametes. The aim of this study was to develop a protocol for hormonal induction of spermiation in the Natterjack toad (Bufo calamita).Materials and methods: Males were divided into three groups ...

ra0001p324 | (1) | WCRB2014

Effects of early exposure to bisphenol A, indole-3-carbinol, and/or genistein and the prostatic development in male rats offspring

Brandt Joyce Zalotti , Pinho Cristiane Figueiredo , Silveira Livia Teresa Ribeiro , Georgete Paulo Cesar , Bozano Jose Eduardo , Barbisan Luis Fernando , Scarano Wellerson Rodrigo

Bisphenol A (BPA) has been investigated for its potential carcinogenic activity for prostate. Studies have shown that indole-3-carbinol (I3C) and genistein (GEN) can activate antiproliferative and proapoptotic signaling pathways. The aim of this study was evaluate the influence of I3C (20 mg/kg) and GEN (5 mg/kg) on the development of prostate of offspring rats exposed during gestation and lactation to BPA (10 mg/kg). Pregnant female Sprague–Dawley rats were divided into ...

ra0001p325 | (1) | WCRB2014

Chemical fingerprinting of embryotoxic mineral oil

Morbeck Dean , Gallacher Christopher , Kalin Robert M

Introduction: Mineral oil is widely used to culture embryos in biomedicine, agriculture, and embryo/stem cell research, yet is poorly defined with significant batch variation causing detrimental effects. The highly hydrophobic nature of this petroleum product attracts polycyclic aromatic hydrocarbons (PAH) and other potentially harmful compounds. Industry requirements for screening oil utilize methods that lack the sensitivity necessary to avoid oil that is overtly embryotoxic...

ra0001p326 | (1) | WCRB2014

Abstract withdrawn....

ra0001p327 | (1) | WCRB2014

Establishment of immortalized neuronal cell lines derived from fetal goat hypothalamus

Suetomi Yuta , Tsukamura Hiroko , Ohkura Satoshi , Matsuda Fuko

Neurons essential for reproductive functions are distributed in hypothalamus. The precise functions of these neurons are not fully analyzed at a cellular level, especially in domestic animals. To analyze cellular mechanisms that control ruminant reproduction, we generated neuronal cell lines by immortalizing fetal goat hypothalamus.The hypothalamic arcuate nucleus was collected from a female Shiba goat at E118. The tissue was dispersed and plated. Twenty...

ra0001p328 | (1) | WCRB2014

Development of novel neurokinin-3 receptor agonists that potently stimulate the GnRH pulse generator

Yamamura Takashi , Misu Ryosuke , Wakabayashi Yoshihiro , Ohkura Satoshi , Oishi Shinya , Fujii Nobutaka , Okamura Hiroaki

Introduction: Pulsatile GnRH secretion is a key determinant for normal gonadal activities. Recent emerging evidence suggests that neurokinin B (NKB) plays a pivotal role in generating GnRH pulse. Here, we aimed to develop novel compounds with potent NKB receptor (NK3R) agonistic activity for future therapeutic applications to improve gonadal functions.Materials and methods: On the basis of the structure-activity relationship study of NKB and tachykinin p...

ra0001p329 | (1) | WCRB2014

Changes in gene expressions induced by perinatal estrogen related to the brain sexual differentiation in rodents

Watanabe Youki , Sakakibara Mototsugu , Uenoyama Yoshihisa , Minabe Shiori , Deura Chikaya , Nakamura Sho , Maeda Kei-ichiro , Tsukamura Hiroko

Brain mechanism responsible for LH surge generation shows sexual dimorphism in rodents. The dimorphism is considered to be due to the perinatal estrogen converted from testicular androgen during developing period. The present paper aimed to clarify the mechanisms mediating the estrogen-induced defeminization of LH surge-generating system. The microarray analysis is an attempt to obtain the candidate gene(s) mediating the perinatal estrogen action causing the brain sexual diffe...

ra0001p330 | (1) | WCRB2014

Metabolic reprogramming may underpin the aetiology of endometriosis

Young Vicky J , Brown Jeremy K , Maybin Jacqueline A , Duncan W Colin , Horne Andrew W

Transforming growth factor beta (TGFβ) can induce The Warburg-effect, or aerobic glycolysis, in tumorigenesis. Energy-rich lactate, a by-product of glycolysis, ‘feeds’ tumour cells and increases apoptosis resistance, angiogenesis, and invasion, fuelling tumour progression and metastasis. As TGFβ is increased in the peritoneal fluid of women with endometriosis, we asked, is there a Warburg-like effect at play in endometriosis.Peritonea...

ra0001p331 | (1) | WCRB2014

Deletion of the C-terminal region of OVGP1 affects porcine IVF

Algarra Blanca , Han Ling , Aviles Manuel , Coy Pilar , Jovine Luca , Jimenez-Movilla Maria

Introduction: The female reproductive tract provides the optimal environment for gamete interaction and embryo development. Understanding the oviductal environment is important for reproducing the in vivo conditions in vitro. OVGP1 is the major non-serum oviductal protein with a variable protein activity depending on its regions. To investigate the potential role of the OVGP1 regions on its activity, full-length (prOVGP1, 527aa) and C-terminal truncated (prOV...

ra0001p332 | (1) | WCRB2014

Androgen receptor-signalling in the male pituitary is dispensable for normal LH secretion but suppresses prolactin production

Ferreira Maria Tedim , Curley Michael , Milne Laura , Smith Lee , O'Hara Laura

Production of testosterone by testicular Leydig cells is under tight regulation by the hypothalamic – pituitary – gonadal (HPG) axis. GnRH, secreted by the hypothalamus, stimulates secretion of LH from the pituitary. LH will then stimulate testosterone production in the Leydig cells, which will feedback to the hypothalamic-pituitary element to inhibit LH secretion.Previously it was accepted that testosterone exerted its negative feedback action...

ra0001p333 | (1) | WCRB2014

Selective expression of two DAZL isoforms in human fetal ovary but not testis

Rosario Roseanne , Anderson Richard

Introduction: Deleted in azoospermia-like (DAZL ) is an RNA-binding protein essential for germ cell entry into meiosis and later stages of germ cell maturation, and its absence is associated with infertility in vertebrates. Efforts to identify in vivo mRNA targets of DAZL have mainly been restricted to mouse, thus human-specific investigations are required.Materials and methods: RT-qPCR and western blotting were used to profile DAZL isoform expr...

ra0001p334 | (1) | WCRB2014

Development of knock-in somatic cells to produce human FGF2 protein on the bovine β-casein gene locus using F2A self-processing peptide

Kim Se Eun , Jeong Young-Hee , Kim Yeong Ji , Kang Man-Jong

Introduction: The production of recombinant protein in transgenic domestic animal is one of the major successes of biotechnology. Knock-in system is a more powerful method to produce mammary gland bioreactor. This study is conducted to development of knock-in somatic cells using bovine β-casein genome for produce of biological substance.Material and methods: The knock-in vector was constructed by using 5.9 kb fragment of upstream of bovine β-ca...

ra0001p335 | (1) | WCRB2014

Neurokinin B activates synchronized intracellular Ca2+ oscillations in KNDy neurons obtained from the hypothalamic arcuate nucleus of Kiss1–GFP transgenic mice

Ikegami Kana , Minabe Shiori , Ieda Nahoko , Goto Teppei , Abe Hitomi , Sanbo Makoto , Hirabayashi Masumi , Maeda Kei-ichiro , Tsukamura Hiroko , Uenoyama Yoshihisa

Pulsatile secretion of GnRH/LH is indispensable for puberty onset and normal reproductive functions in mammalian species. A cohort of neurons expressing three neuropeptides, kisspeptin, neurokinin B (NKB), and dynorphin (KNDy neurons), localized in the hypothalamic arcuate nucleus (ARC), are considered to be a source of GnRH pulse generator. A synchronous discharge of KNDy neurons might be obligatory for pulsatile GnRH secretion. The present study aimed to determine whether NK...

ra0001p336 | (1) | WCRB2014

Autophagy supports the survival of the conceptus obtained by assisted reproductive technologies

Toschi Paola , Czernik Marta , Zacchini Federica , Loi Pasqualino , Ptak Grazyna Ewa

Assisted reproductive technologies (ARTs) are generally considered to be safe, but recent studies suggest an excess of negative pregnancy outcomes. The mechanisms responsible for adverse events share similar condition like abnormal placentation, which finally leads to impaired embryo growth due to limited nutrient uptake. High autophagy activation was reported in placentae obtained from normal and compromised pregnancies. Given the evidence of placental abnormalities in pregna...

ra0001p337 | (1) | WCRB2014

Abstract withdrawn....

ra0001p338 | (1) | WCRB2014

Gonadotropin-independent follicle development in the Kiss1/− female rats

Shirota Mariko , Kawashima Jun , Shirota Kinji , Uenoyama Yoshihisa , Nakamura Sho , Ikegami Kana , Maeda Kei-ichiro , Tsukamura Hiroko

Introduction: Kisspeptin, encoded by Kiss1, is a neuropeptide that directly regulates GnRH secretion from the hypothalamus. The present study aimed to examine a relationship between the circulating gonadotropin levels and follicle development using the Kiss1 knockout (Kiss1 KO) adult female rats.Materials and methods: Kiss1 KO and WT animals were obtained by mating of Kiss1+/− males and female...

ra0001p339 | (1) | WCRB2014

Hypothalamic arcuate nuleus-specific enhancer for kisspeptin expression of female mice

Goto Teppei , Tsukamura Hiroko , Tomikawa Junko , Abe Hitomi , Fukanuma Tatsuya , Imaura Takuya , Takase Kenji , Sanbo Makoto , Tomita Koichi , Hirabayashi Masumi , Maeda Kei-ichiro , Uenoyama Yoshihisa

Two populations of kisspeptin neurons, located in the arcuate nucleus (ARC) and anteroventral periventricular nucleus (AVPV), are considered to be involved in generating GnRH pulse and surge respectively. The present study aimed to determine the region-specific enhancer for ARC Kiss1 gene expression by in vivo reporter assay using transgenic (Tg) mice. Three GFP reporter constructs (long-, middle-, and short-length) were generated by insertion of GFP cDNA in ...

ra0001p340 | (1) | WCRB2014

Proteome-wide changes in liver function by continuous exposure to sewage sludge

Filis Panagiotis , Turner Emily Eaton , O'Shaughnessy Peter , Fowler Paul A

Introduction: A complex cocktail of poorly biodegradable chemicals is ubiquitous in the modern environment. Exposure to such chemicals contributes to diseases such as metabolic syndrome and infertility. Since the liver is the primary defence organ against xenotoxicants, we analysed the liver proteome of sheep continuously exposed (from pre-conception until early adulthood) to sewage sludge. Our aim was to identify dysregulated pathways and understand how a chemical cocktail ca...

ra0001p341 | (1) | WCRB2014

Determination of expression of MFGE8 in porcine oviduct and uterus

Guillen-Martinez Ascension , Nicolas Carla Moros , Acuna Omar Salvador , Rico Maria Jose Izquierdo , Aviles Manuel

Introduction: Billions of sperm are deposited in female tract during porcine insemination but only a few thousands are able to reach the oviduct to facilitate fertilization. It has been reported that some of the sperm membrane proteins involved in fertilization are secreted by epididymis (i.e. SPAM-1, CRISP, and MFGE8). It was observed that the mouse oviduct is able to secrete some of these proteins. Some of them are implicated in fertilization since sperm fertility of KO mice...

ra0001p342 | (1) | WCRB2014

Mechanisms underlying pituitary microvascular remodelling in thoroughbred horses during the annual reproductive cycle

Yeomans Angus , Thompson Nichol , Castle-Miller Jennifer , Bates David O , Tortonese Domingo

In photoperiodic species, remodelling of pituitary microvasculature appears to play a key role in the regulation of the annual reproductive cycle. Recent studies have revealed seasonal changes in the vascular loops of the ovine pituitary stalk. Microvasculature remodelling is under the control of pro-angiogenic (VEGF165a) and anti-angiogenic (VEGF165b) isoforms of vascular endothelial growth factor (VEGF). Here, we examined the differential expression of ...

ra0001p343 | (1) | WCRB2014

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ra0001p344 | (1) | WCRB2014

Lack of gonadotropin release in Kiss1 knockout male rats

Uenoyama Yoshihisa , Nakamura Sho , Hayakawa Yuki , Ikegami Kana , Deura Chikaya , Minabe Shiori , Tomikawa Junko , Goto Teppei , Ieda Nahoko , Sanbo Makoto , Tamura Chihiro , Hirabayashi Masumi , Maeda Kei-ichiro , Tsukamura Hiroko

Kisspeptin, encoded by Kiss1 gene, has attracted attention as a key neuropeptide in controlling reproduction in mammals. Earlier studies with Kiss1 or its cognate receptor Gpr54 knockout (KO) mice showed the indispensable role of kisspeptin–GPR54 signaling on reproduction, but detailed analysis of gonadotropin release were still limited. Here we have generated Kiss1 KO rats to evaluate a role of kisspeptin on pulsatile gonadotropin secr...

ra0001p345 | (1) | WCRB2014

The gender-specific hormone INSL3 demonstrates inter-fetal transport of hormones between male and female fetuses

Anand-Ivell Ravinder , Vernunft Andreas , Poeppel Caterina , Ivell Richard

Introduction: The peptide hormone insulin-like peptide 3 (INSL3) is a unique product of the developing fetal testis that can be detected at a very early stage of pregnancy in fetal blood and also in the surrounding amniotic fluid from where bidirectional exchange of components can occur as the fetal skin is still not keratinized. To explore inter-fetal transport processes, the pig, as a multiparous species, provides an excellent model.Materials and metho...

ra0001p346 | (1) | WCRB2014

Does xenotoxicant-disrupted fetal sheep thyroid development persist into adulthood?

Filis Panagiotis , Ramona Lauma , Murray Graeme , Hombach-Klonisch Sabine , Klonisch Thomas , O'Shaughnessy Peter , Fowler Paul A

Introduction: A complex cocktail of poorly biodegradable chemicals is ubiquitous in the modern environment and exposure to such chemicals contributes to diseases including thyroid dysfunction. The thyroid gland regulates growth and function of multiple organs and we previously showed that exposure to xenotoxicants (via maternal grazing on sewage sludge fertilised pastures) disturbed fetal sheep thyroid development. Our aim here was to assess whether the effects seen in fetal s...

ra0001p347 | (1) | WCRB2014

The role of kisspeptin for defeminization and masculinization of sexual behaviors in rats

Nakamura Sho , Uenoyama Yoshihisa , Ikegami Kana , Tamura Chihiro , Sanbo Makoto , Hirabayashi Masumi , Tsukamura Hiroko , Maeda Kei-ichiro

Kisspeptin, encoded by Kiss1 gene, is considered to be a key regulator for GnRH secretion. Kiss1 knockout (KO) rats are infertile due to the absence of gonadotropin secretion in both sexes. Kiss1 KO male rats receiving testosterone during their adult age showed no male sexual behaviors (mount, intromission, and ejaculation), but showed female sexual behavior (lordosis) in Kiss1 KO males. To examine whether kisspeptin is also needed to induce...

ra0001p348 | (1) | WCRB2014

Mating decreases the sulfated HNK1 carbohydrate motif over a 130 kDa glycoprotein in the rat endosalpynx

Zuniga Lidia M , Nunez Teresa A , Andrade Juan C , Croxatto Horacio B , Orihuela Pedro A , Morales Patricio

Mating is a source of signals to the female reproductive tract to ensure reproductive success. Previously, we reported that mating induces the expression of carbohydrate sulfotransferase 10 (CHST10) in the rat endosalpynx. This enzyme participates in the sulfation of the carbohydrate motif named human natural killer 1 (HNK1), synthesized over membrane proteins and lipids. Both molecules, CHST10 and HNK1, are involved in cell-to-cell interactions. Herein, we determined the effe...

ra0001p349 | (1) | WCRB2014

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ra0001p350 | (1) | WCRB2014

Effects of cooling and cryopreservation on sperm mitochondrial membrane and sperm motility

Sicherle Carmen Cecilia , de Paula Freitas-Dell'Aqua Camila , Ackermann Camila Louise , Derussi Ana Augusta Pagnano , Mothe Gabriele , de Souza Fabiana Ferreira , Papa Frederico O , Lopes Maria Denise

The aim of this study was to analyze the effects of refrigeration and cryopreservation on dog sperm special on sperm motility and mitochondrial membrane potential (MMP). A total of 15 ejaculates, first and second fractions, were collected from five dogs. Semen was diluted (80×106 sptz/ml) on Tris-egg-yolk medium with 8% of glycerol (one step), filled into 0.5 ml French straw and refrigerated at 5 °C for 1 h. After straws were suspended 6 cm above liquid ni...

ra0001p351 | (1) | WCRB2014

Defining the role of sry-related HMG box 4 in male differentiation in vivo

Hartwig Sunny

Mammalian gonads arise in both sexes from bilateral urogenital ridges that have the potential to develop as ovaries or testes. In mice, the genital ridges are first evident at embryonic day (E) 9.5. At E10.5, a critical switch in gonad development occurs in which XY gonads express the testis-determining gene (sex-determining region Y chromosome) Sry. Sry belongs to the SOX family of nuclear transcription factors. Male...

ra0001p352 | (1) | WCRB2014

Granulosa cells from human small antral follicles changes gene expression during culture

Jeppesen Janni V , Petersen Tonny Studsgaard , Borgbo Tanni , Andersen Claus Yding

Background: As a consequence of cryopreservation of human ovaries preformed in our laboratory we have access to human small antral follicles.Material: Human granulosa cells (GC) from individual small antral follicles (4–6 mm in diameter) were spilt in two and either cultured or snap-frozen immediately after isolation from the follicular fluid. Microarray analysis was used on both the cultured and the non-cultured sample, to examine their gene expres...

ra0001p353 | (1) | WCRB2014

Effects of common FSH receptor single-nucleotide polymorphisms on the follicular fluid hormone profile and the granulosa cell gene expression in human small antral follicles

Borgbo Tanni , Kluckova Hana , MacEk Milan , Chrudimska Jana , Jeppesen Janni V , Lindgren Ida , Giwercman Yvonne Lundberg , Hansen Lise Lotte , Andersen Claus Yding

Background: FSH is one of the key hormones of mammalian reproduction. FSH signaling may be affected by single-nucleotide polymorphisms residing in the FSH receptor (FSHR) gene. The most pronounced effects of these polymorphisms are likely to be displayed in the follicle fluid, which acts as a reservoir for granulosa cell secreted hormones induced by FSH.Methods: Genetic polymorphisms of FSHR in position -29 (FSHR -29) and in codon 307 and codon ...

ra0001p354 | (1) | WCRB2014

Telomerase reverse transcriptase is an epithelial progenitor marker in the mouse endometrium

Deane James A , Breault David T , Gargett Caroline E

Rare epithelial and stromal stem/progenitor cells (SPC) have been identified in the mouse endometrium. However the role of these SPC in endometrial regeneration is unclear due to the lack of a traceable marker. We used transgenic mice with reporter constructs for the stem cell marker telomerase reverse transcriptase (mTert) to identify and characterize mouse endometrial SPC. Endometrial mTert expression was examined by microscopy and flow cytometry in mice expressing GFP under...

ra0001p355 | (1) | WCRB2014

Cdh1- and Cdc20-independent APC activity in mouse oocytes

Nabti Ibtissem , Carroll John

Introduction: The anaphase-promoting complex (APC) is an E3-ubiquitin ligase responsible for regulated destruction of substrates at specific stages of the cell cycle. Two APC co-activators, Cdc20 and Cdh1, mediate the timing and selectivity of substrates recognition. Progression through meiosis in oocytes utilises the same molecular players, although the start–stop nature of the female meiosis invokes additional levels of regulation. Recently, we reported that Cdk1 and MA...

ra0001p356 | (1) | WCRB2014

In vitro differentiation of spermatogonial stem cells to functional sperm in zebrafish

Sakai Noriyoshi

Introduction: Spermatogenesis is a complex process in which spermatogonial stem cells (SSCs) self-renew, develop to differentiated spermatogonia, and then give rise to meiotic spermatocytes. Functional sperm are ultimately differentiated from postmeiotic haploid spermatids. Cell culture methods representing this process will facilitate analyzing molecular function and imaging molecules. Here we describe consecutive two culture methods by which zebrafish SSCs enable to differen...

ra0001p357 | (1) | WCRB2014

Effects of maternal peri-conception and first trimester protein supplementation on placental development

Edwards Jennifer L , Copping Katrina J , Alibhai Aziza , Graham Holly , Barnett Stephanie , Robinson Robert S , Perry Viv E A , Mongan Nigel P , Rutland Catrin Sian

Introduction: This study investigates the effect of dietary crude protein on the cellular composition of the 98 days bovine placenta.Materials and methods: Primiparous 14-month-old Bos. indicus cross heifers in Australia (n=350) were individually fed isocalorific high (H; 14%) or low (L; 7%) CP from day −60 prior to conception to 23 days post conception (dpc). At 23 days post-conception the two groups were further split into high ...

ra0001p358 | (1) | WCRB2014

High testicular size is related with low intramale variation of sperm design in red deer

Ros-Santaella Jose Luis , Pintus Eliana , Garde Jose Julian

Introduction: Testes investment plays a major role in male reproductive success because larger testes are expected to produce higher sperm number and quality. In birds, comparative studies have shown that testis size – a proxy measure of postcopulatory sexual selection – is negatively related with the intraspecific variation of sperm design. By contrast, links between the intramale variation of sperm morphometry and testicular size in mammals are unknown. Here, for t...

ra0001p359 | (1) | WCRB2014

Improvement of cloned blastocyst quality upon CHIR treatment of donor cells

Czernik Marta , Oikawa Mami , Iuso Domenico , Ogura Atsuo , Loi Pasqualino

Somatic cell nuclear transfer (SCNT) has a broad spectrum of many potential applications but the efficiency is still disappointingly low. It is known that major cause of it, is incorrect or incomplete reprogramming of the transferred donor nuclei.Many report indicates that Wnt/β-catenin signaling pathway plays a pivotal role in the process of somatic cell reprogramming. As CHIR is known as one of the activators of Wnt pathway by inhibiting glycogen ...

ra0001p360 | (1) | WCRB2014

The capacity to reprogramme is an intrinsic property of adult somatic cells

Iuso Domenico , Toschi Paola , Anzalone Debora Agata , Czernik Marta , Ptak Grazyna Ewa , Loi Pasqualino

Oct4 and Nanog are the transcription factors responsible for pluripotency maintenance in embryonic stem cells (ESCs), and activation of these genes in differentiated cells is considered the earliest signal of nuclear reprogramming. To date, nuclear reprogramming in somatic cells can be induced by nuclear transfer, induced expression of defined factors (Oct4, c-myc, Sox-2, and Kfl4) and, more recently, by a short exposure to a low pH. However, work in salamander as well as in p...

ra0001p361 | (1) | WCRB2014

The role of cell adhesion molecules in primordial follicle formation

Pepling Melissa , Burton Joshua , Senderoff Dana

Introduction: Oocytes develop in cysts of interconnected cells that undergo breakdown and become surrounded individually by granulosa cells to form primordial follicles. Mechanisms controlling cyst breakdown and primordial follicle formation are not well understood but cell adhesion molecules (CAMs) may a play role. Our model is that CAMs keep oocytes in cysts and must be down regulated for primordial follicles to form. A second set of CAMs would then be necessary for interact...

ra0001p362 | (1) | WCRB2014

Nuclear distribution of genotoxic damage in rainbow trout (Oncorhynchus mykiss) sperm after cryopreservation

Gonzalez-Rojo Silvia , Fernandez-Diez Cristina , Lombo Marta , Rodriguez Vanesa Robles , Paz Herraez Maria

Introduction: Sperm DNA is prone to suffer damage that could have a negative impact on the progeny development. Moreover genes packaged in different chromatin blocks or located in different nuclear areas, could have differential sensitivity to genotoxic agents. Our group has demonstrated that in rainbow trout, with the nucleus homogeneously compacted with protamines – lacking histones –, the distribution of damage is dependent on the genotoxic agent. The present work...

ra0001p363 | (1) | WCRB2014

Conceptus-derived proteins in bovine uterine luminal fluid at the time of pregnancy recognition in cattle

Forde Niamh , Bazer Fuller W , Spencer Thomas E , Lonergan Patrick

Maternal recognition of pregnancy (MRP) in cattle requires secretion of sufficient interferon tau (IFNT) by the conceptus to prevent luteolysis. However, recent evidence suggests that additional conceptus-derived products may be involved. The aim of this experiment was to identify conceptus-derived proteins in uterine luminal fluid (ULF) during MRP. Following estrous synchronization, the protein content of ULF from cyclic (n=4) and confirmed pregnant (n=4) he...

ra0001p364 | (1) | WCRB2014

Live imaging reveals novel dynamic events in transitioning mouse gonocytes

Jarvis Sheba , Fraser Scott E , Winston Robert M , Readhead Carol W

Introduction: Early developmental events in the testis set the scene for future spermatogenesis. After birth, gonocytes, the precursors to type-A spermatogonia reproliferate and become transiently motile in a poorly understood process. Aberrations are associated with altered fertility or pre-malignant states. Gonocytes exist as overlapping subpopulations at varying developmental stages, which makes them difficult to study.Materials and methods: Transgeni...

ra0001p365 | (1) | WCRB2014

The impact of DNA damage induced by sperm cryopreservation in Xenopus

Morrow Sean , Guille Matt , Holt William , Garner Trent

Introduction: This project aims to investigate the nature of cryo-induced DNA damage in the spermatozoa of the model species Xenopus tropicalis. We have turned to Xenopus as a tool to achieve this because the genome is available and we aim to identify specific genomic regions that are most affected during sperm cryopreservation.Materials and methods: The sperm chromatin dispersion (SCD) test was used to show DNA damage and the potent DN...

ra0001p366 | (1) | WCRB2014

Sperm total motility and membranes integrity after selection by centrifugation in colloid solutions of ram cryopreserved semen

Gomes Bergstein Tacia , Cassia Bicudo Luana , Rodello Leandro , Romualdo Weiss Romildo , Dimas Bicudo Sony

Introduction: Frozen and thawed ovine semen undergo morphological and functional changes that prevent or decrease the efficiency of fertilization. Sperm selection methods seek to improve the quality and viability of the fertilizing materials.Material and methods: Four sperm selection methods by centrifugation were employed, using two silica colloidal solutions coated with silane (Ovipure™– Nidacom, Gothenburg, Swedis) or by polyvinylpyrrolidone...