(1)

Human sperm use different behaviours for different aspects of sperm transport and fertilisation. We have investigated the effects of preparation method (density gradient method and direct swim-up) on sperm behaviour and manipulated Ca2+ signalling (the primary regulator of sperm motility) to induce different behaviours and assessed their effects on penetration of human sperm through viscous medium (methylcellulose).Cells prepared by swim-up pe...

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ra0002p002 | (1) | SRF2015

Associations between circulating non-esterified fatty acids and uterine function in postpartum dairy cows

Cheng Zhangrui , Hiu Yiu Yao Veronica , Wathes D Claire

Evidence suggests that excessive non-esterified fatty acid (NEFA) production during postpartum negative energy balance (NEB) increases the risk of uterine infection. The aims of our study were to investigate associations between circulating NEFAs and uterine gene expression in postpartum dairy cows with NEB using microarray and bioinformatics techniques. Mild NEB (n=6) and severe NEB (n=6) in 12 cows were produced using different milking and...

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ra0002p003 | (1) | SRF2015

Characterization of migratory and gonadal porcine PGCs

Zhang Haixin , Cheng Kiu Wai , Alberio Ramiro

Primordial germ cells undergo an orchestrated developmental program prior to establishing the mature gametes. Most of our knowledge on PGC development stems from studies in mice, however recent studies in humans revealed important differences in the mechanisms of PGC development. Since pigs share many embryological features with non-rodent species, like humans, and human PGC can only be studied from early gonadal stages, we have used pig embryos to study the developmental prog...

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ra0002p004 | (1) | SRF2015

Effect of macrophages on steroidogenesis and cell migration in bovine ovaries

Samir Moafaq S , Mattar Dareen S , Knight Phil G

Introduction: Cyclic ovarian function involves continual tissue remodelling. During ovulation macrophages invade the ovary and secret pro-inflammatory cytokines such us TNF- and IL6 that have local actions on ovarian cells. Here we investigated i) the effect of macrophages on secretion of oestradiol by granulosa cells (TC) and androstenedione by theca cells (TC) and ii) the effect of macrophages on TC and stroma cell (SC) migration was also assessed using an in vitro ...

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ra0002p005 | (1) | SRF2015

A deep RNA sequencing study of mammalian sperm RNA: identifying common cross-species expression motifs indicating functionality

Nadj Stefanie , Huntriss John , Miller David

Despite being transcriptionally silent,1 mature spermatozoa contain small amounts of RNA comprising various classes and although, de novo translation of spermatozoal RNA has been demonstrated,2 the role of these RNAs is currently unclear. However, looking across species boundaries and comparing the RNA profiles of sperm from a number of mammals may give insights into their function in the process of reproduction, including potentially post-fertil...

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ra0002p006 | (1) | SRF2015

Endometrial level of lysophosphatidic acid (LPA) and expression of LPA receptors mRNA in endometrosis in the mare

Szostek Anna , Karasinska Beata , Ferreira-Dias Graca , Skarzynski Dariusz Jan

Lysophosphatidic acid (LPA) is a bioactive lipid mediator that exerts a wide range of biological actions. LPA acting specifically through LPA receptor 1 (LPAR1) seems to be essential for the development of fibrosis in several organs. We determined whether the LPALPARs signalling system correlates to equine endometrial fibrosis (endometrosis). A total of 24 uteri from diestrus (n=6 for each category: I, II A, II B, and III according to Kenney and Doig classificat...

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ra0002p007 | (1) | SRF2015

Effect of hyaluronan on some molecular markers of implantation in sheep endometrial cells in vitro

Raheem Kabir Ayobami , Fouladi-Nashta Ali A

Previous studies indicate a pattern of expression for hyaluronan (HA) synthases in the reproduction system regulated by steroid hormones which produce different HA sizes with diverse physiological functions. The objective of the present study was to determine the effect of low molecular weight (LMW) and high molecular weight (HMW) HA on molecular markers of embryo implantation using in vitro model of endometrial cell culture system. Sheep endometrial cells at confluen...

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ra0002p008 | (1) | SRF2015

Expression of porcine CXADR and its role in blastocyst formation

Kwon Jung-Woo , Kim Nam-Hyung , Choi Inchul

Coxsackie virus and adenovirus receptor, CXADR (CAR) is a member of the tight junction protein (TJP also known as JAM) family of adhesion receptor and located on a cytoplasmic membrane surface of intercellular tight junctions. CXADR are reported to be expressed during preimplantation in human embryos, but its function in early embryo development was not investigated. In the present study we determine temporal and spatial expression patterns of CXADR in porcine embryos and inve...

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ra0002p009 | (1) | SRF2015

A model of tissue-engineered (3d) decidua to study the effects of environmental pollutants on endometrial physiology

Mannelli Chiara , Szostek Anna , Letta Francesca , Łukasik Karolina , Jankowska Katarzyna , Piotrowska-Tomala Katarzyna , Paulesu Luana Ricci , Skarzynski Dariusz Jan

The increasing exposure to environmental chemicals is a burden for human reproduction, and can alter endometrial functions and maternalembryo interactions. The available in vitro endometrial models often fail to represent the complexity of cellular environment, cells shape and organization, and could give misleading results on the effects of environmental chemicals. Here, a tissue-engineered (3D) decidua was used as a toxicological model. Human stromal cells wer...

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ra0002p010 | (1) | SRF2015

Characterisation of the dog as a sentinel species for human exposure to endocrine disrupting chemicals

Sumner Rebecca , Byers Andrew , Taylor Oliver , Moxon Rachel , White Natasha , Craigon Jim , Zhang Zulin L , England¹ Gary C W , Lea Richard

Introduction: A temporal decline in canine male fertility parallels that reported in the human. Our hypothesis is that this is associated with exposure to environmental chemicals (ECs). We investigated i) a relationship between canine testicular chemical profiles and Sertoli cell numbers and ii) the effects of testicular concentrations of chemicals on sperm DNA fragmentation, motility and vitality.Methods: Canine adult testes (routine castrations) from t...

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ra0002p011 | (1) | SRF2015

Assessment of major reproductive problems and reproductive status of crossbred (Holstein Friesian X Zebu) dairy cattle in and around Mekelle, Tigray, Ethiopia

Mekonnin Alemselam Birhanu , Harlow Chris , Howie Forbes , Gidey Goitom , Tadesse Desalew , Desta Gidena , Ashebir Gebregiorgis , Gebrekidan Berihu , Gugsa Tadesse , Riley Simon

Ethiopia maintains an extensive livestock population, however reproductive performance of cattle and their breeding management are unsatisfactory. Currently, the sole diagnostic tool in the country is rectal palpation, which is inaccurate for early pregnancy, and causes embryonic and fetal loss. This study assessed major reproductive problems using questionnaire survey, and trialled simple, cost-effective alternative monitoring approaches using on-farm diagnostic tools to dete...

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ra0002p012 | (1) | SRF2015

Expression and possible role of receptor-interacting protein kinase (RIPK) 1 and 3 in the bovine CL

Hojo Takuo , Szostek Anna , Jonczyk Agnieszka , Łukasik Karolina , Piotrowska-Tomala Katarzyna , Okuda Kiyoshi , Skarzynski Dariusz Jan

Programmed necrosis or necroptosis is an alternative form of cell death that is regulated by a caspase-independent pathway. The aim of the study was to determine if necroptosis participate in bovine CL luteolysis. In Experiment 1. the RIPK1 and RIPK3 mRNA transcription was determined in bovine i) CLs from early, developing, mid, late and regressed-stage (n=4 for each stage), ii) CLs after colpotomy collected within 0-, 2-, 4- and 12-h (each n</em...

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ra0002p013 | (1) | SRF2015

Whether equine Corpus Luteum is a site for LPA synthesis and/or a target for LPA action?

Łukasik Karolina , Szostek Anna , Staszkiewicz Joanna , Skarzynski Dariusz Jan

Lysophosphatidic acid (LPA) is a simple phospholipid and exerts various biological functions effecting on reproductive processes in rats, pigs, ewes and cow. LPA acts through three subtypes of Edg family G protein-coupled receptors (LPA1-3) and non-Edg family LPA receptors (LPA4-6). The aim of the study was to determine LPA: i) concentrations and its receptors mRNA transcription, and ii) effect on P4 secretion from equine CL during the oestrous cycle and...

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ra0002p014 | (1) | SRF2015

The effect of E.coli lipopolysaccharide (LPS) on bovine luteal endothelial cell network formation and steroidogenesis in vitro

Mohammed Zeravan A , Mann George E , Robinson Robert S

In dairy cows, post-partum uterine inflammation caused by Gram negative bacteria (e.g. E. coli) suppresses follicular function and reduces fertility. LPS is an endotoxin that is present on outer membrane of Gram-negative bacteria. Furthermore, LPS is detected in follicular fluid (0.040.88 μg/ml) of cows with endometritis. This study tested the hypothesis that LPS would decrease the formation of luteal endothelial cell (EC) network and progesterone ...

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ra0002p015 | (1) | SRF2015

The association between uterine disease and subsequent reproductive performance in commercial UK dairy herds

Mohammed Zeravan A , Hudson Chris D , Robinson Robert S , Mann George E

Dairy cow fertility has declined over the past 50 years coinciding with increased milk production. Uterine health is an important factor with endometritis prevalent in high-yielding dairy cows. This study assessed the association between uterine disease on reproductive performance in 78 commercial UK dairy herds. Data from 59 118 lactations (n=29 157 cows) was collected from 2000to 2009 and included presence of uterine disease, calving date, and insemination informati...

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ra0002p016 | (1) | SRF2015

Membrane potential is crucial for one of the [Ca²⁺]_i oscillations profiles induced by progesterone in human spermatozoa

Nitao Elis Torrezan Goncalves Ramalho , Publicover Stephen J

Ca2+ signalling is critical for regulation of sperm motility. [Ca2+]i oscillations, which may underlie observed switching of sperm behaviors, occur in human spermatozoa stimulated with progesterone. Our work aimed to investigate the potential contribution of changes in membrane potential, leading to cyclical activation of voltage dependent Ca2+-influx pathways, to [Ca2+]i oscillations. Spermatozoa we...

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ra0002p017 | (1) | SRF2015

Differential proteomic profiles of porcine follicular fluid associated with a high fibre diet and later fertility

Jarrett Selene , Ferguson Elizabeth M , Kurian Dominic , Gill Andy C , Ashworth Cheryl J

In a previous study, following IVF, oocytes from gilts fed a high fibre diet for the first 19 days of their third oestrous cycle, produced blastocysts with more cells than oocytes from control-fed gilts. We hypothesise that FF protein composition is altered by the diet and that this confers the reproductive benefits.The current study compared the protein composition of pooled Day 19 FF from 12 high fibre-fed pigs and 12 control-fed pigs in search of biom...

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ra0002p018 | (1) | SRF2015

An E4BP4 knockout model to assess the distribution of uterine Natural Killer cell subsets in mouse pregnancy

Peet Claire Joy , Boulenoaur Selma

Natural Killer cells have been linked to a number of disorders of pregnancy by both mouse studies and association studies in humans, with NK cell dysfunction leading to reduced spiral artery remodelling. However, little research has centred on the importance of the newly divided subsets of uterine Natural Killer cells, some of which appear to develop independently of E4BP4, in spiral artery remodelling, and subsequently fetal growth. Hence, we sought to analyse the dependence ...

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ra0002p019 | (1) | SRF2015

PCR detection and phylogenetic analysis of bovine herpesvirus 4 field-isolates from dairy cows in Thailand

Swangchan-Uthai Theerawat , Suadsong Siriwat , Techakumphu Mongkol

Introduction: Postpartum dairy cows are susceptible to uterine infections that leading to poor fertility. During this period the uterine defence mechanisms were compromised by poor energy status and the preceding viral infections, e.g. bovine herpesvirus 4 (BoHV-4) resulting in uterine pathology initiated by microorganisms. BoHV-4 has been isolated from healthy cows and cows experienced with mastitis, metritis and endometritis worldwide.Methods:...

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ra0002p020 | (1) | SRF2015

The effect of oocyte-specific ablation of N- and O-glycans on the cumulus extracellular matrix

Lo Belinda K M , Archibong-Omon Agnes A , Ploutarchou Panayiota , Williams Suzannah A

Each egg, when ovulated from a follicle, is surrounded by cumulus cells. Prior to ovulation, these cumulus cells secrete cumulus extracellular matrix (cECM) molecules, resulting in cumulus expansion. Cumulus expansion has been linked to the developmental quality of the oocyte. Hyaluronan (HA), the major constituent of the cECM, is stabilised by molecules such as heavy chains (HCs), pentraxin 3 (PTX3) and tumour necrosis factor-stimulated gene 6 (TSG6) during expansion. All of ...

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ra0002p021 | (1) | SRF2015

Inter-generational effect of maternal analgesic exposure on female ovaries of second generation of female rats

Wang Yili , MacPherson Sheila , McKinnell Chris , van den Driesche Sander , Sharpe Richard

Analgesics which work through altering the production and/or actions of prostaglandins (PGs) are widely used by pregnant women. In earlier research, the analgesics in maternal blood circulation can target cyclooxygenase-2 and PGE2 receptors in fetal germ cells (GC), affect the next generation. Both sexes of F1 rats exposed to analgesic in utero showed reduced germ cell (GC) number in gonads at the fetal stage, and reduced ovarian oocyte reserve in adult stage and/or i...

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ra0002p022 | (1) | SRF2015

Role of sperm thiols? redox status in keeping rat sperm quiescent in cauda epididymis

Yadav Santosh Kumar , Kumar Lokesh , Pandey Aastha , Kushwaha Bhavana , Maikhuri Jagdamba Prasad , Gupta Gopal

Background: The sperm require energy-intensive motility to reach the female gamete for delivering the male genome. This energy is conserved by keeping the sperm quiescent in cauda epididymis before ejaculation, but the molecular mechanisms controlling this unique process remain an enigma. Cauda sperm produce H2O2, and we have attempted to study the redox regulation of sperm motility.Methods: Quiescent and motile sperm were collected...

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ra0002p023 | (1) | SRF2015

The effects of environmental chemicals on bovine luteal function

Thompson Robert , Quint George , Robinson Robert S , Lea Richard G , Zhang Zulin L , Woad Kathryn J

Dairy herd fertility is in decline and poses a significant economic problem, with associated animal welfare concerns. One factor that may contribute to this decline is exposure to environmental chemicals (ECs) some of which have endocrine disrupting activity and have been linked to declining fertility in other species. We hypothesised that ECs are present in bovine ovarian tissue at levels able to perturb luteal development and function. The tissue content of DEHP and a panel ...

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ra0002p024 | (1) | SRF2015

Ovarian transcriptome profile before, during and after the onset of premature ovarian failure in a mouse with oocyte-specific deletion of Mgat1 and Cgalt1 genes

Galaz Heidy Kaune , Williams Suzannah A

Premature ovarian failure (POF) affects ~1% of women over 40 and is idiopathic in 7490% of cases. A transgenic mouse model of POF has been generated (known as double mutant; DM) resulting from oocyte-specific deletion of two glycosyltransferases. Glycoproteins from DM oocytes lack complex O- and N-glycans. DM females are fertile at 6-weeks, infertile by 9-weeks and exhibit POF by 12-weeks of age with follicle depletion, elevated gonadotropins and decrea...

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ra0002p025 | (1) | SRF2015

CG9879 in the Drosophila testis

Cole Rebecca Louise , Jiang Jianqiao , Caporilli Simona , White-Cooper Helen

In Drosophila, the testis meiotic arrest complex (tMAC) regulates the expression of a vast number of genes involved in spermatogenesis and is essential for entry into male meiosis. Among its many targets are a few putative transcription factors that could further regulate spermatogenesis genes. CG9879 is implicated as one such tMAC-induced transcription factor as it is testis-specific, tMAC-dependent, and homologous to the known transcription factors TBP,...

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ra0002p026 | (1) | SRF2015

Regulation of expression of oocyte quality control factor, ATRX, in bovine oocytes during maturation

O'Shea Lynne C , Hensey Carmel , Fair Trudee

In humans, mutation of the ATRX gene leads to improper methylation of repetitive DNA sequences. By performing a meta-analysis on published microarray data across several model species we previously identified ATRX as a potential biomarker of oocyte quality. The aim of the present study was to determine the expression and regulation of ATRX at a protein level, in the bovine cumulus oocyte complex (COC).ATRX protein was found to be expressed during oogenes...

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ra0002p027 | (1) | SRF2015

Dynamics of hyaluronic acid (hyaluronan) binding proteins in relation to sperm capacitation and the acrosome reaction

Torabi Forough , Miller David

Objective: Considering the recent interest in the use of hyaluronan binding as a sperm selection marker for ICSI, it is becoming increasingly important to revisit and characterise the sperm hyaluronic acid binding proteins (HABPs) that are an integral part of the selection process. To this end, the HABPs in non-capacitated, capacitated and acrosome reacted spermatozoa were investigated.Design: A laboratory based investigation using immune-cytochemical an...

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ra0002p028 | (1) | SRF2015

Expression of coxsackie virus and adenovirus receptor (CXDAR) affects tight junction complex ADN cell lineage in mouse blastocyst

Juong Yelin , Yu Dae-yul , Choi Inchul

Coxsackie virus and adenovirus receptor (CXADR also known as CAR), a tight junction component molecule was reported to be expressed in epithelial cells. Previous studies reported that CXADR play an important role in tight junction complex. However, the role of CXDAR in blastocyst formation has not been investigated. Here we demonstrated that transcript levels of CXADR were elevated at eight-cell stage onward and highest at the blastocyst. To investigate the biological function...

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ra0002p029 | (1) | SRF2015

Successful isolation, culture and karyotyping of equine placental cells from failed early pregnancies

Rose Belinda Vivienne , Cabrera-Sharp Victoria , Cameron Ian , Crabtree James , Crowhurst James , Firth Marvin , Ghosh Sharmila , McGladdery Andrew , Neal Huw , Pynn Jan , Pynn Oliver , Raudsepp Terje , Smith Charlie , Verheyen Kristien , Wathes D Claire , Wise Zara , de Mestre Amanda

Early pregnancy loss (EPL) in the mare is defined as loss of pregnancy between initial detection and day 65 of gestation. It occurs in 710% of pregnancies and yet little is known about the underlying pathologies. A lack of suitable conceptus material has limited investigation into the role of chromosomal defects in EPL. The objective of this study was to develop a method to isolate and culture placental cells isolated from failed pregnancies to enable further genetic cha...

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ra0002p030 | (1) | SRF2015

The endocrine disrupting chemicals bisphenol A, dichlorodiphenyltrichloroethane (DDT), methoxyclor and ethinylestradiol modulate thecal steroidogenesis in vitro

Cheewasopit Warakorn , Knight Phil G

Introduction: Previous reports in different species have shown that various endocrine disrupting chemicals (EDCs) can modulate ovarian steroidogenesis with the majority of studies focussing on granulosa cells. To test the hypothesis that exposure to EDCs might also perturb thecal steroidogenesis we conducted in vitro doseresponse experiments to evaluate the direct effects of selected EDCs on androstenedione and progesterone production by cultured bovine theca ce...

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ra0002p031 | (1) | SRF2015

Does maternal progesterone supplementation in early pregnancy affect fetal development?

Mareckova Magda , Siemienowicz Katarzyna Joanna , Wang Yili , Rae Mick , Duncan W Colin

Some adult diseases are programmed in utero by fetal exposure to abnormal concentrations of steroid hormones. Threatened miscarriage in early pregnancy is treated by progesterone in many countries although robust evidence of efficacy is lacking. We hypothesised that increased progesterone concentrations may alter fetal development. In a small pilot study, using a pregnant sheep model, we administered 200 mg progesterone twice weekly from d20d75 of gestation and ...

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ra0002p032 | (1) | SRF2015

PGF2α-PTGFR signaling promotes angiogenesis in the porcine endometrium during early pregnancy

Kaczynski Piotr , Baryla Monika M , Kowalewski Mariusz P , Waclawik Agnieszka

Introduction: Prostaglandin F2α (PGF2α) is mainly known to be involved in luteolysis. However, our recent studies indicate that PGF2α synthesis and its receptor (PTGFR) expression are up-regulated in the porcine endometrium during embryo implantation. Aims of present study were: i) to immunolocalize PTGFR protein in uterus; ii) to elucidate the involvement of PGF2α-PTGFR signaling in angiogenesis in the porcine endometrium.Methods: Se...

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ra0002p033 | (1) | SRF2015

Functional evaluation of miRNAs during the ovarian follicular/luteal transition

Mohammed Bushra T , Duncan W Colin , Donadeu Francesc X

Little is known about the involvement of miRNAs in luteal development. Cattle provide a convenient model to study ovarian physiology in monovular species. Our previous microarray studies in bovine showed significant upregulation of miR-96 and miR-132 in luteal relative to follicular tissues. In the present study we used an in vitro model of forskolin-induced follicular granulosa cell luteinisation, transfection with specific locked nucleic acid inhibitors or mimics of...

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ra0002p034 | (1) | SRF2015

Effect of cryopreservation on follicular development in human ovarian tissue

Getreu Natalie , Morwood Tom , Young Martin , Fuller Barry , Hardiman Paul J

Introduction: Recent progress in cancer therapy has significantly decreased mortality rates. However, these therapies whilst curative can cause significantly reduced fertility or sterility. Current fertility preservation methods are limited to women who either have a partner or who will use donor sperm and without a hormone sensitive tumor. Ovarian tissue transplantation could offer an alternative for these patients. Unlike other studies, we assessed the impact of cryopreserva...

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ra0002p035 | (1) | SRF2015

Differential gene expression in granulosa–lutein cells from women with polycystic ovaries is independent of the dose of FSH given for ovarian stimulation

Velupillai Jocelyne , Lerner Avi , Christopoulos Georgios , Liyanage Maneshka , Lavery Stuart , Trew Geoffrey , Laird Mhairi , Mora Jocelyn M , Tsui Victoria , Hardy Kate , Franks Stephen

Polycystic ovary syndrome (PCOS) affects over 5% of women of reproductive age. It is associated with an ovulatory infertility, menstrual disturbances as well as metabolic abnormalities including obesity and insulin resistance. PCOS has a strong genetic basis but studies of ovarian gene expression in PCOS are limited by the difficulty in obtaining tissue samples from women with and without PCOS. For that reason, most studies have been performed in granulosalutein (GL) cel...

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ra0002p036 | (1) | SRF2015

Effects of maternal peri-conception and first trimester protein supplementation on circulating progesterone levels and concomitant conception rates in yearling heifers

Copping Katrina J , Callaghan Matthew , McMillen Caroline , Rodgers¹ Ray , Perry Viv E A

Introduction: This study investigates the effect of dietary crude protein on conception in range beef heifers and uses circulating progesterone as a measure of early embryonic loss known to be a major cause of reproductive inefficiency.Methods: Primaparous 14 month old Bos indicus cross heifers (n=350) were selected from a range population. 60 days prior to artificial insemination they were randomly assigned to two equal groups and indi...

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ra0002p037 | (1) | SRF2015

In vitro culture of IVM derived porcine embryos: comparisons of single-step and sequential media systems on embryo quality and cryosurvivability

Bartolac Louise K , Sjoblom Cecilia , Grupen Christopher

In pigs, the quality of In vitro-produced embryos is much lower than that of in vivo-derived embryos. The poor quality of In vitro-produced porcine embryos has been attributed to the apparently sub-optimal embryo culture media that is commonly created in-house. Few studies have examined the capacity of commercially available media, typically used to culture human embryos, to support the development of porcine embryos. The aim of the present study was...

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ra0002p038 | (1) | SRF2015

Experimental models for challenging the ‘KNDy hypothesis’: the acute response of GnRH secretion to nutrition

Rietema Stacey E , Hawken Penny A R , Smith Jeremy T , Lehman Michael N , Martin Graeme B

In the KNDy hypothesis, the latest concept explaining the control of GnRH secretion, the pulsatile GnRH signal is controlled by arcuate nucleus cells that produce kisspeptin (K), neurokinin B (N) and dynorphin (Dy). The interplay among these peptides involves one of them acting as a pace-setter, one as a brake, and the other as communicator of the final signal to the GnRH cells. To date the KNDy hypothesis has proven to be robust, but, if it...

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ra0002p039 | (1) | SRF2015

Poster withdrawn....

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ra0002p040 | (1) | SRF2015

Sex specific effects of maternal dietary protein during the periconceptional period upon hepatic gene expression in heifer progeny

Morrison Janna , Copping Katrina J , Hernandez-Medrano Juan H , Rodgers Raymond , McMillen Caroline , Perry Viv E A

Sex specific expression occurs in 2030% of all hepatic genes in the adult animal (1). We have shown that there is an interaction between gender and the level of maternal dietary protein intake on bovine fetal development as early as 3639 days post conception (dpc) We hypothesise that exposure to different levels of maternal protein intake in the periconceptional period (60d preconception to 23dpc) and in the postconceptional period (2398dpc) will result in a ...

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ra0002p041 | (1) | SRF2015

Regulation of CyclinD2 by Smad3 and Foxl2 during early follicle development

Granados-Aparici Sofia , Sharum Isam , Hardy Kate , Franks Stephen , Waite Sarah , Chapman Neil , Fenwick Mark

Introduction: Primordial follicles are relatively quiescent structures that form the basis of the ovarian reserve. Maintenance of the quiescent state, and conversely, release from this state towards irreversible growth, involves mechanisms that are currently unresolved. Two transcription factors have been implicated alongside this process in granulosa cells (GCs). Specifically, Smad3 was recently identified in the nuclei of GCs in small single-layered follicles whereas Foxl2 i...

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ra0002p042 | (1) | SRF2015

Genome-wide cDNA microarray screening to correlate gene expression profiles of aged mouse ovary to the size of the reserve of the primordial follicle

Yuki Emika , Kanno Shou , Morimoto Motoko , Inoue Masayasu , Kobayashi Jin

Size of the reserve of primordial follicles in mammalian ovary is critical for female reproduction and reproductive senescence. Despite an accurate estimation of the size of the reserve of primordial follicles remains unestablished, due to lack of molecular markers. We performed to explore the expressed gene in the ovarian correlated with primordial follicle in posterior reproductive duration using a wide array in mouse. ICR mouse bread until 9 or 58 weeks was used (9W and 58W...

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ra0002p043 | (1) | SRF2015

Melatonin-induced regulation of vascular endothelial growth factor (VEGF) isoform expression and microvasculature remodelling in the pituitary gland of seasonally anoestrous ewes

Ferreira Catarina Fernandes , Mooney Josephine , Castle-Miller Jennifer , Bates David O , Clarke Iain , Tortonese Domingo

Recent studies have shown that microvasculature remodelling of the ovine pituitary gland is seasonally regulated through mechanisms involving the differential expression of pro-angiogenic (VEGF165) and anti-angiogenic (VEGF165b) isoforms of vascular endothelial growth factor (VEGF). As for other photoperiodic species, the pattern of pineal melatonin secretion mediates the effects of day length on the sheep seasonal reproductive cycle. In this study, we in...

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ra0002p044 | (1) | SRF2015

Melatonin regulates seasonal variations in prolactin and follicle stimulating hormone synthesis via alternative splicing of pituitary vascular endothelial growth factor-A

Castle-Miller Jennifer , Bates² David O , Tortonese Domingo

The pars tuberalis (PT) of the ovine pituitary contains high-density melatonin receptors (MT1) known to participate in the seasonal regulation of prolactin (PRL) output from lactotroph cells in the pars distalis (PD). However, the paracrine mechanism relaying the signal from the PT to the PD remains unresolved. We have recently shown that MT1-positive cells in the PT co-express vascular endothelial growth factor-A isoforms (VEGF165/VEGF165b), and that VEG...

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ra0002p045 | (1) | SRF2015

Periovulatory oxygen levels within the porcine oviduct obtained by laparoendoscopic single-site surgery

Albors Octavio Lopez , Sanchez Hurtado Miguel , Gutierrez Hugo Andres , Margallo Francisco Sanchez , Gadea Joaquin , Latorre Rafael

This work was aimed at determining in vivo levels of oxygen (O2) within the porcine oviduct. 13 gilts (G) and eight sows (S) were used. A left lateral paralumbar laparoendoscopic single-site surgical approach (GelPOINT Advanced, SingleMedical→) was carried out under CO2 pneumoperitoneum. Laparoscopy manoeuvres allowed pulling up the ovary towards the single-site port and upon visual inspection pigs were assorted into preovulatory (PreO) or ...

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ra0002p046 | (1) | SRF2015

Effect of Rhodobacter sphaeroides LPS on toll-like receptor 4 in bovine endometrial cells

Chotimanukul Sroisuda , Swangchan-Uthai Theerawat , Techakumphu Mongkol

Introduction: Endometritis is one of the most common reproductive diseases affected postpartum bovine uteri. Lipopolysaccharide (LPS) from Gram-negative bacteria is recognized by Toll-like receptor 4 (TLR4). Binding of LPS and TLR4 mediates the expression of proinflammatory cytokines and chemokines. The excessive inflammatory responses could be involved with endometrial damage.TLR4 antagonist Rhodobacter sphaeroides LPS (RsLPS), is a potent antagonist of LPS from path...

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ra0002p047 | (1) | SRF2015

Waddlia chondrophila stimulates CXCL8 expression in ruminant trophoblast cells via p38 and p42/44 MAPK dependent pathways

Wheelhouse Nick M , Storrie Skye , Vacca Francesco , Barlow Peter , Rae Mick , Longbottom David

Background: Waddlia chondrophila (W. chondrophila) is an emerging abortifacient pathogen which has been identified in the placentae of humans and cattle. The organism is a member of the order Chlamydiales, and shares many similarities at the genome level, and in growth studies, with other well-characterised zoonotic chlamydial abortifacients, such as Chlamydia abortus (C. abortus). We have previously observed significant responses i...

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ra0002p048 | (1) | SRF2015

The effect of prenatal exposure to androgen and to a high fat diet on obesity in female mice

Laird Mhairi , Hardy Kate , Franks Stephen

Polycystic ovary syndrome (PCOS) is a complex endocrine disorder that presents in women of reproductive age. The aetiology of PCOS is poorly understood, however it is likely that women with this disorder are predisposed to produce excess androgen at or well before puberty. Environmental and lifestyle factors then contribute to the multifarious symptoms observed. One such key external factor is diet. These studies utilised a prenatally exposed androgen mouse model to investigat...

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ra0002p049 | (1) | SRF2015

Does kisspeptin exert a local modulatory effect on ovarian steroidogenesis?

Mattar Dareen S , Cheewasopit Warakorn , Samir Moafaq , Knight Phil G

Introduction: Kisspeptin, a neuropeptide secreted in the hypothalamus and encoded by the Kiss-1 gene, has a role in promoting the release of GnRH and LH in various species. However, the possibility that kisspeptin exerts additional peripheral actions at the level of the bovine gonad has not been investigated. The current aims were to investigate whether: i) kiss-1 and its receptor (GPR54) are expressed in the bovine ovary; ii) kisspeptin or kisspeptin antagonist ca...

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ra0002p050 | (1) | SRF2015

Immunohistochemical detection of hypoxya markers in the porcine oviduct

Albors Octavio Lopez , Aparicio Monica , Llinares Anabel , Candanosa Eugenia , Parraga Ester , Ayala M Dolores , Boronat Talia , Gutierrez Hugo Andres , Latorre Rafael

This work was aimed at localizing and evaluating the expression of endogenous hypoxia markers in the porcine oviduct. Oviduct samples were obtained from reproductive tracts of sows (n=20) in the slaughterhouse. Upon visual inspection of the ovary the samples were allocated into late follicular (LF), early luteal (EL) and late luteal (LL) phases of the estrous cycle. Ampulla and isthmus sections were stained against the following specific primary antibodies: HIF2α, VEGF, F...

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ra0002p051 | (1) | SRF2015

SPRASA knockout mice are sub-fertile

Prendergast Deborah , Liu Bonnia , Sheling Andrew Neil , Chamley Larry

Introduction: SPRASA is a protein, that is expressed in the acrosome of sperm, as wel as on the oolemma of many mammals, which we identified as the target of antisperm antibodies from some infertile men. In order to further understand the importance of SPRASA we produced knockout mice and examined the effect of this gene knockout on murine fertility.Methods: With ethics approval (AEC_R811) knockout mice were generated lacking the expression of exons 4 an...

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ra0002p052 | (1) | SRF2015

The effect of reproductive ageing on chromatin configuration and amino acid metabolism in germinal vesicle staged sheep oocytes in vitro

Topipat Chutima , Lu Jianping , Fernandez Esther Collado , Huntriss John , Picton Helen

Reproductive ageing in females is associated with reduced oocyte maturation potential and developmental competence. Oocyte chromatin configuration and metabolism, have recently been established as correlates of oocyte developmental competence in a number of species. Here, we have evaluated the effect of reproductive ageing on oocyte chromatin morphology and amino acid turnover in germinal vesicle (GV)-staged, ovine oocytes.Cumulusoocyte complexes w...

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ra0002p053 | (1) | SRF2015

Investigating the action of 1,25(OH)₂D₃/vitamin D₃ in the human ovary: relevance to fertility and polycystic ovary syndrome (PCOS)

Standing Andrew , Georgiou Christiana , Bano Gul , Mason Helen Diane , Rice Suman

In recent years it has become apparent that vitamin D3 (VD) has a fundamental role in reproductive function, with deficiency of the hormone being implicated in several reproductive pathologies: endometriosis, pre-eclampsia and PCOS. VD is essential for oestrogen synthesis in both males and females; via indirect mechanisms of calcium homeostasis but also direct regulation of aromatase expression. Women with PCOS are more likely to suffer from VD deficiency, with an inverse corr...

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ra0002p054 | (1) | SRF2015

Characteristics of bovine granulosa cells cultured under low and high oxygen tensions in the presence of different concentrations of melatonin

Zeebaree Bayar Kabeer , Kwong Wing Y , Mann George E , Gutierrez Carlos G , Sinclair Kevin D

Previous studies have assessed the effects of melatonin on cultured granulosa cells under 5% CO2 in air, and focused on its antioxidant properties. Effects of melatonin under more physiological oxygen tensions are not known. Here we cultured granulosa cells from antral follicles (46 mm diameter) in fibronectin-coated 12-well plates (at 600 000 cell/ml) in TCM199, with melatonin added at one of four concentrations (0, 20, 200, 2000 pg/ml) under either i) low ox...

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ra0002p055 | (1) | SRF2015

Impact of maternal age on oocyte amino acid turnover and mitochondria DNA copy number in sheep

Fernandez Esther Collado , Campbell Bruce K , Lu Jianping , Picton Helen

Introduction: Reproductive ageing is associated with a reduced ovarian reserve and suppression of oocyte developmental competence (quality) as evidenced by increased meiotic segregation errors, epigenetic alterations and reduced fertilisation rates. Metabolic indicators of oocyte quality such as oocyte amino acid profile (AAP) and mitochondria DNA (mtDNA) copy number may provide insights into the mechanism of declining oocyte quality with age. This study investigated the effec...

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ra0002p056 | (1) | SRF2015

Serine/threonine kinase receptor associated protein (Strap) inhibits early follicle development in mouse ovaries

Sharum Isam , Granados-Aparici Sofia , Fenwick Mark

Introduction: The molecular mechanisms involved in regulating growth of small, gonadotrophin-independent follicles are poorly understood. We have previously shown that the canonical TGFβ signalling intermediate, Smad3 is highly expressed in granulosa cells (GCs) of single-layered follicles. Furthermore, a reduction in the overall expression of Smad3 is associated with the onset of multi-layering and increased granulosa cell proliferation, suggesting modulation of TGF&#946...

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Society for Reproduction and Fertility Annual Conference 2015

BiosciAbstracts