Reproduction, Fertility and Development - the Society for ...

Reproduction, Fertility and DevelopmentAn international journal for the publication oforiginal work, review and comment in the fields ofreproductive biology, reproductiveendocrinology and development biology, including puberty, lactation and fetal physiology when they fall within thesefields. The Journal will accept papers dealing with biochemistry, cell biology, molecular biology, endocrinology, immunologyand genetic sex determination of animals including humans, but only if these are related to reproduction, fertility ordevelopment.This journal is one ofthe Australian Journals ofScientific Research published by CSIRO PUBLISHING in cooperation withthe Commonwealth Scientific and Industrial Research Organization (CSIRO) and the Australian Academy of Science.Editorial policy for the series is developed by a Board of Standards appointed jointly by CSIRO and the Academy ofScience.CSIRO PUBLISHING requires that all authors of a multi-authored paper agree to its submission. The Journal has used itsbest endeavours to ensure that work published is that of the named authors except where acknowledged and, through itsreviewing procedures, that any published results and conclusions are consistent with the primary data. It takes no responsibilityfor fraud or inaccuracy on the part ofthe contributors.Editorial Advisory CommitteeAcceptance ofpapers is the responsibility ofthe Managing Editor in consultation with the Editorial Advisory Committee onbehalfofthe Board of Standards. Committee members may advise the Managing Editor on the selection ofreferees and mayadjudicate in the case of conflicting or adverse reports. All papers are peer reviewed.Chairperson:G. B. Martin, University ofWestern AustraliaMembers: D. T. Armstrong, University ofAdelaideS. G. Matthews, University ofToronto1. E. Harding, University ofAuckland1. Hinds, CSIRO Sustainable Ecosystems, Canberrap. 1. Kaye, University of Queensland, BrisbaneW M. C. Maxwell, University of SydneyM. F. Pera, Monash Institute ofReproduction and Development, MelbourneAdvisory PanelA. M. Carter (Denmark); 1. R. G. Challis (Canada); Y. Combarnous (France); B. Fadem (USA); A. P. F. Flint (UK); W W Hay(USA); K. M. Henderson (New Zealand); 1. 1. Guillette (USA); 1. Huhtaniemi (Finland); B. Jegou (France); P. M. Motta(Italy); T. Nagai (Japan); E. Nieschlag (Germany); C. Pholpramool (Thailand); 1. D. Skinner (South Africa); 1. F. Smith (NewZealand); D. G. Whittingham (UK); R. Yanagimachi (USA)Managing Editor:Subscription rates for 2002Institution print & online:Institution online only:Personal print & online:Personal online only:C. M. MyersAustralian and New Zealand customers pay in AU$. All othercustomers pay in US$. Prices include air freight delivery.Special rates are available for society members. Visit:www.publish.csiro.auljournals/rfd8 issues per year; 4 archival print issues plus supplement.ISSN 1031-3613© CSIRO Australia 2002.A$$550$480$120$115US$$625$550$150$115The Instructions to Authors are published in the first issue ofeachvolume. Copies are available on request. Information about theJournal, including the Instructions to Authors, is available onWorld Wide Web at URLhttp://www.pubIish.csiro.au/journals/rfdAll inquiries and manuscripts should be forwarded to:The Managing EditorReproduction, Fertility and DevelopmentCSIRO PUBLISHINGPO Box 1139 (150 Oxford Street)Collingwood, Victoria 3066, AustraliaInternational telephone 613 9662 7629; facsimile 613 9662 7611Email: publishing.rfd@csiro.auSOCIETY FORREPRODUCTIVE BIOLOGYPROGRAM AND ABSTRACTSTHIRTY-THIRDAnnual Scientific Meeting22-25 September 2002Adelaide Convention Centre, Adelaide, SAFivnt cover images. Main photograph: injection of DNA into the pronucleusofa centrifuged porcine zygote; courtesy ofAndrew French, Centre for EarlyHuman Development, Institute of Reproduction and Development, MonashUniversity. Inset: newborn tammar wallaby, Maeropus eugenii, attached to theteat in the pouch; courtesy ofLyn Hinds and CSIRO Sustainable Ecosystems.4:~i~1~- ~C SIR 0 Academy of SciencePermission to photocopy items from this journal is granted byCSIRO to libraries and other users registered with the CopyrightClearance Center (CCC), provided that the appropriate fee foreach copy is paid directly to the CCC, 222 Rosewood Drive,Danvers, MA 01923, USA. For copying in Australia, CSIROPUBLISHING is registered with Copyright Agency Limited,Level 19, 157 Liverpool Street, Sydney, NSW 2000. Specialrequests should be addressed to the Managing Editor,Reproduction, Fertility and Development, PO Box 1139 (150Oxford Street), Collingwood, Victoria 3066, Australia.Copyright © Society for Reproductive Biology, 2002ISSN 1031-3613

The Society for Reproductive Biology(SRB)acknowledges support of the following 2002 Conference Sponsors:Society forReproductive Biology (SRB)PEST ANIMAL CONTROL CRCCRC FOR INNOVATIVEDAIRY PRODUCTSTABLE OF CONTENTSPrevious SRB/ASRB Office Bearers/Awardees 4CRCCORPORATE SPONSORSNovo Nordisk Pharmaceuticals(Satchel and Dinner Sponsor)GlaxoSmithKline (Dinner Sponsor)GlaxoSmithKline2002 SRB Office Bearers 5SRB Student Information 6Adelaide Convention Centre Floorplan and DelegateInformation 7Trade Delegate Information 10Eli Lilly (Welcome Function Sponsor)Joint SRB/ESA Program Details 15Serono (Junior Scientist Award Sponsor)(SeroncVSRB Program - Session Details 33Abstracts (in order of presentation) 43Reproduction, Fertility and DevelopmentAuthor Index 93Joint SRB/ESA Program at a Glance 96CONFERENCE SECRETARIATAustralian Science Network Pty Ltd (ASN)Unit 21 B Balnarring Village Centre(PO Box 200) Balnarring, VIC 3926Tel: +61 3 5983 2400 Fax: +61 3 5983 2223Email: mp@asnevents.net.au23

Society for Reproductive BiologylAustralian Society for Reproductive BiologySociety for Reproductive BiologyPast Office Bearers & AwardeesGODING /JUNIORYEAR CHAIRMAN TREASURER SECRETARY FOUNDERS SCIENTISTLECTURE WINNER 2002 Office Bearers1969-701970-71RG Wales JR GodingTJ Robinson1971-72 Chairman John Hearn1972-73 Honorary Secretary Rob Gilchrist1973-74Honorary TreasurerChris OINeiliIG White BM Bindon1974-75IACummin Committee Members Graham Jenkin (Deputy Chair)CWEmmens1975-76 WHansel Peter Kaye1976-77 DT Baird Kate Loveland1977-78TD GloverCamilla Myers (ad hoc)GM Stone IACumming1978-79CH T ndale-BiscoeAnne TurnerDM de Kretser1979-80GMH ThwaitesPostgraduate Representative Christine White1980-81 GHMcDowell KPMcNattRJ ScaramuzziCommun. OfficerlWebmaster Jim Cumminsl Michael Sinosich1981-82BK Follett PJ LufenJSA Committee Chair Alan Tilbrook1982-83 NWMoore RJ Rodger &BG Miller J Wilson SRB Secretariat Australian Convention and Travel ServiceTK RobertsCBGow1983-84BM Bindon SP FlahertJKFindlay1984-85 A Bellve C O'NeillJMCummins1985-86 CD NancarrowBM Bindon1986-87 WHansel LJ WiltonG Evans1987-88HG Bur er A Sto'anoff Chair Kate LovelandBP Setchell LA Hinds1988-89FW Bazer MB HarveMembers Arun Dharmarajan1M Shelton1989-90 GD Thorburn AHTorne Ann DrummondJC Rodger GB Martin1990-91 RMMoor HMassa Sarah MeachemPLKaye1991-92CR Austin DKGardner Sarah RobertsonRJ Scarammuzzi CG Tsonis1992-93JK Findlay SWWalkden- Andrew SinclairLABrownSalamonsenAnne Turner1993-94 GCLi ins CMMarkeAO Trounson LJ Wilton1994-95 I Huhtaniemi MJ Hotzel & SMPHedgerMcDou all1995-96RF Seamark JD Curlewis1996-97No Lecture S Robinson1997-98JGThompson F Bronson M Jas erMB RenfreeChair Ray RodgersMB Harvey1998-99 DM de Kretser M Panteleon Members Rob Gilchrist2000-01 R Short C Gargett Wendy IngmanJHeam PKaye T Lavranos D Albertini WIngman Hamish Eaton2001-02 R Gilchrist A TrounsonTony RobertsJeremy ThompsonESA Program Organising Committee Chair: Leon BachBP Setchell BJWaddellSRB Program Organising Committee 2002RF Seamark I van Wezel Local Organising Committee (ESAlSRB) 20024 5

SOCIETY FOR REPRODUCTIVE BIOLOGYStudent MembersINFORMATION FOR DELEGATES AND PRESENTERSThere are three events at the 2002 SRB Annual Conference that students should not miss!All events are being held on Monday 23 rd September.RIVE.RIU\.HKCARPARKfESTIVAL DRIVE1pm Student Lunch Lecture Adelaide Convention CentreProfessor Alan Trounson will be participating in a roundtable discussion specifically for students,following his Founders Lecture. All students are welcome to attend, and a light lunch will beprovided.7pm SRB Students Annual Meeting University ofAdelaide Staff ClubThis is a great opportunity to meet fellow students working in the field of reproductive biology, andvoice your opinions on how we can improve your experience of SRB student membership. Themeeting will be held right before the Student BBQ at the University of Adelaide Staff Club.7.30pm SRB/ESA Student BBQ University ofAdelaide Staff ClubAs students, we know the value of a cheap meal and night out! For only $30 per person, theSRB/ESA Local Organising Committee is providing a full buffet dinner, drinks and entertainment.Donlt miss this chance to meet and have a great night with other SRB and ESA students, andnetwork with other conference delegates working in your field.WANT TO BECOME AN SRB STUDENT MEMBER?For only $35 per year, membership gives you:• Member registration discount at SRB Annual Scientific Meetings• Free SRB Abstract Booklets and Newsletters• Eligibility for SRB and RFD Travel Awards and the Junior Scientist Award• Reduced purchase price on: Reproduction, Fertility and Development and Reproduction• Access to a professional network in the field of Reproductive BiologyPlease see your Postgraduate Student Representative, Christine White, for an applicationform. Email: christine.white@med.monash.edu.auNORTH TERRACELEVEL ONE//-- -"/ "",,-'-.../ (/ \iHVATT )OO"COUOT Il'MW.__------------') ~-----"REPRODUCTION, FERTILITY & DEVELOPMENTTravel Award 2003Presented annually to an SRB member PhD student or junior postdoc with the best eligiblemanuscript published in RFD in the year prior to 31 st March 2003 (application closing date).Comprises $1000 towards overseas conference travel and a year's subscription to RFD.Applications forms at http://www.publish.csiro.au/journals/rfdMore info at Students Annual Meeting.The Speaker Preparation RoomConference Room 3 is being used as the speaker preparation room throughout the event.Networked computers in this room will allow MS Powerpoint presentations loaded here to beshown in any of the session rooms. Technicians and assistants will be in attendance in the room,and speakers are encouraged to load their presentations as soon as possible to avoid last minuterushes. Carousels will also be in this room for the pre-loading of any 35mm slides but these mustbe reserved in advance, as it is not the standard or preferred means of presentation.6 7

Abbott Laboratories Site Nos: 73&74KURNELL NSW 2230Phone: 02 9668 1850www.abott.comAbbott Laboratories is a global diversifiedcompany dedicated to the discovery,development, manufacture and marketing ofhealthcare products and services. Abbott withReductil, a new weight management drug waslaunched into the Australian market in February2002, and Gopten, an ACE Inhibitor, are pleasedto be associated with the ESA & SRB and ADS &ADEA conferences 2002.Alphapharm Site Nos: 50-53GLEBE NSW 2037Phone: 02 9298 3953Fax: 02 9566 4686www.alphapharm.com.auAlphapharm is one of Australia's leadingpharmaceutical companies. Alphapharm develop,manufacture, market and distribute prescriptionand pharmacy-only medicines. While Alphapharmspecialises in bringing patent-expired medicinesto market, which allow people save money,Alphapharm have a pipeline of innovativeproducts in development. Recent introductionsinto the Australian market include Campral, Bicorand Diabex 1000. Alphapharm is part of MerckKGaA.Astrazeneca Pty Ltd Site Nos: 7&8NTH RYDE NSW 2113Phone: 02 9978 3882Fax: 02 99783730www.astrazeneca.comAstraZeneca is one of the world1s leadingpharmaceutical companies. It has a strongresearch base and an extensive product portfoliodesigned to meet the medical needs of a broadrange of patients. The current AZ CardiovascularPortfolio includes Atacand, Zestril, Plendil,Betaloc, Tenormin, Inderal and Imdur. A majorresearch and development effort is currentlyunderway aiming to bring to market exciting newtherapies in the areas of hypercholesterolaemiaand thrombosis. AstraZeneca is committed tofinding tomorrow's medical cures.CONFERENCE TRADE EXHIBITORSAtCor Medical Pty Ltd Site No: 17WEST RYDE NSW 2114Phone: 02 9874 8761Fax: 02 9874 9022www.atcormedical.comAtCor Medical market the innovative SphygmoCorrange of cardiovascular diagnostic devices. TheSphygmoCor system allows you to non-invasivelydetect diabetic patients who are at risk of heartattack and stroke. Now you can see, monitor andmanage the key central processes driving thecardiovascular risk of your patients.Aventis Pharma Site Nos: 41-49LANE COVE NSW 2066Phone: 02 9422 6416Fax: 02 9422 6418www.aventis.comAventis Pharma is one of the world1s leadingpharmaceutical companies. With a strong focuson Diabetes, Aventis Australia has invested over$16 Million in the past 2 years to diabetesresearch alone, and has planned to contributeanother $15Million in the next 2yrs again solely tothe prevention and management of diabetes.Bristol Myers Squibb Site No: 10.2NOBLE PARK VIC 3174Phone: 03 9213 4000www.bms.comBristol-Myers Squibb is a leading diversifiedworldwide health care company with the missionto develop and market innovative products thatextend and enhance human life. The company isa leading maker of innovative therapies forcardiovascular, metabolic and infectious diseases,central nervous system and dermatologicaldisorders, and cancer.DSL Australia Pty Ltd Site No: 21BAULKHAM HILLS NSW 2153Phone: 02 9680 7200Fax: 0296807255www.DSLabs.comDSL (Diagnostic Systems Laboratories) is arecognized leader in endocrine diagnosticsinvolved in the development, manufacture anddistribution of immuno-diagnostic test kits. DSLmanufacture and distribute approximately 120products which can be classified into the follOWingareas: androgen assessment, fertility andreproductive function, disease markers, thyroidfunction, bone and mineral metabolism, growthfactors, metabolism, and infectious diseases.Eli Lilly Site No: 25-30WEST RYDE NSW 2114Phone: 02 9325 4486Fax: 02 9325 4573www.lilly.comEli Lilly and Company is a leader in diabetesresearch and has been since producing the firstcommercially available insulin in 1923. This yearEli Lilly introduced Actos (pioglitazone) toAustralia, a type 2 diabetes treatment that targetsinsulin resistance. Also included in the diabetesportfolio is the Humalog range of rapid actinginsulin analogs. Eli Lilly Australia is part of theglobal Eli Lilly and Company, a research-basedpharmaceutical corporation dedicated to creatingand delivering innovative health care solutionsaimed at improving quality of life. Eli Lilly Australiawas established in 1960, employs 403 people andinvests more than $22.7 million annually inAustralian research in the critical disease states inwhich it operates; neuroscience, oncology,infectious diseases, endocrinology, includingosteoporosis and diabetes, and cardiovasculardisease.Genzyme Australasia Site No: 9PO Box 6207, Baulkham Hills Business CentreBAULKHAM HILL BIC NSW 2153Phone: 02 9680 8383Fax: 0296342777www.genzyme.com.auThyrogen® (thyrotropin alfa - rch) is arecombinant form of TSH developed for use inthyroid cancer patients that allows patients toremain on thyroid hormone suppression therapythroughout the course of periodic testing forrecurrence of cancer or metastases, therebyavoiding the debilitating symptoms ofhypothyroidism associated with thyroid hormonewithdrawal.GlaxoSmithKline Site Nos: 58-63BORONIA VIC 3155Phone: 03 9721 4314Fax: 039721 4333www.gsk.comGlaxoSmithKline (GSK) Australia is one ofAustralia1s largest pharmaceutical and healthcarecompanies. GSK has four main sites in Australia,employing more than 1500 people. It is Australia'slargest vaccine manufacturer and a leadingsupplier of medicines for asthma, bacterial andviral infections, depression, migraine,gastroenterological disease, epilepsy, smokingcessation and pain relief. More than 16 millionAustralians rely on at least one of GSK'smedicines, vaccines or consumer healthcareproducts. The company invests more than $25million in R&D each year, making it one ofAustralia's top 20 R&D investors. GSK Australiaplays a significant role in the globalpharmaceutical supply chain - exporting 60% ofproduction to more than 50 countries, with totalexport earnings totalling $197 million in 2001.IPSEN Pty Ltd Site No: 11MT WAVERLEY VIC 3150Phone: 03 9550 1843Fax: 03 9545 3513www.ipsen.com.auIpsen's strong history of creative and successfuldrug discovery illustrates its commitment to thedevelopment of highly specialised products tomeet specific needs in areas such as neurologyand endocrinology. An international companybased in Europe, Ipsen represents a relativelynew name in the Australian pharmaceuticalindustry and currently markets Somatuline® LA(Ianreotide) - a somatostatin analogue indicatedfor the treatment of acromegaly. Somatuline LAprovides an alternative in somatostatin analoguesand has been used extensively in the UK andEurope over recent years.1011

Mayne Pharma Site No: 13PARKVILLE VIC 3052Phone: 03 8341 5000Fax: 03 8341 5050www.faulding.comMayne's pharmaceutical division is represented inmore than 50 countries worldwide, with a focus ondeveloping, manufacturing and marketing genericinjectable and oral pharmaceuticals, primarily forthe hospital market. Mayne Pharma Australia hasalso established a global reputation for marketinginnovative patent protected in-licensed products.Through our specialised care team we marketproducts in the following therapeutic areas;Transplant Medicine, Oncology, Endocrinology,Urology, Infectious Diseases, Neurology andPsychiatry.Merck Sharp & Dohme Site No: 12SOUTH GRANVILLE NSW 2142Phone: 02 9795 9185Fax: 0297959602www.msd-australia.com.auMerck Sharp & Oohme (MSO) Australia is aresearch based pharmaceutical company and hasbeen looking after the health of Australians since1952, a history of which MSD is extremely proud.MSD invests a considerable amount intoAustralian research and development. Over 90%of MSD products are manufactured and packagedin Australia, which leads to new investment andinfrastructure opportunities. The ongoingcommitment of MSO in Australia is evidenced byits achievement in becoming the largestpharmaceutical exporter in the country, exportingto more than 16 countries throughout the world.Novartis Pharmaceuticals Site No:10NTH RYDE NSW 2113Phone: 02 9805 3555Fax: 02 9888 3408www.pharma.novartis.comNovartis is a global life sciences company formedin 1997 through the merger of Sandoz and Ciba.In Australia, Novartis Pharmaceuticals is theleading supplier of pharmaceutical products tohospitals. The company boasts a broad portfolioof p~oducts in the fields of neurology,transplantation, oncology, endocrinology andgastroenterology. Novartis has recently increasedits focus on the areas of Oncology,Transplantation and Opthalmics by the formationof business units to manage these specialtyproducts. The company is also committed toextensive clinical research within AustralianInstitutions. Some well known brands of theNovartis Oncology business unit include Femara® (Ietrozole), Sandostatin LAR® (octreotide),Zometa® (zoledronic acid) and the recentlydeveloped tyrosine kinase inhibitior, GliveC®(imatinib). The Novartis representatives presentat this meeting would be happy to answer anyquestions related to Novartis Oncology products.NovoNordisk PharmaceuticalsSite Nos: 1-6 & 64-72PO Box 6086PARRAMATTA BUSINESS CENTRE NSW2150Phone: 02 9890 0723Fax: 02 9683 3029www.novonordisk.com.auNovo Nordisk recognizes the commitment thatmembers of the Endocrine Society of Australiamake towards patients with diabetes andendocrinological disease. The Novo Nordiskcorporate goal is to improve the way people withdiabetes live and work. We continue to be at theforefront of diabetes discovery and are committedto the development of new and innovativetreatments, in partnership with sophisticatedpatient focused delivery systems. Our goal is tomeet the individual needs of patients byrecognizing that all patients are not the same andrequire different solutions to the everydaychallenges they face.Pfizer Pty Ltd Site Nos: 31-34WEST RYDE NSW 2114Phone: 02 9850 3333Fax: 02 9850 3146www.pfizer.comPfizer is the worldls leading research based healthcare company. They discover, develop,manufacture and market innovative medicaltreatments for humans and animals. Ranked asone of the worldls top 10 companies, Pfizer willinvest US$5 billion in research and developmentfor conditions including heart disease, arthritis anddiabetes. Pfizer is dedicated to improving themanagement of people with CV risks andassociated complications.Pharmacia Site Nos: 18&19RYOALMERE NSW 2116Phone: 02 9848-3150Fax: 02 9848 3333www.pharmacia.comPharmacia Corporation is committed to improvinghealth and wellbeing for people around the worldand is recognised as an innovative leader in anumber of therapeutic areas, includingarthritis/inflammation, gastroenterology, CNS,ophthalmology, oncology, neurology, women'shealth, pain management, infectious diseases andmetabolic diseases. Pharmacia Australia isinvolved in the research, and development of newdrugs for the company and is passionate in itscommitment to providing the highest qualityproducts to foster good health and wellnessthroughout the community.Sanofi-Synthelabo Site No: 24LANE COVE NSW 2066Phone: 02 9937 1706Fax: 02 9937 1755www.sanofLcom.auResulting from a series of mergers with severalpharmaceutical companies, Sanofi-Synthelabo isa leading pharmaceutical group with itsoperational head office in France. It is one of thenew French companies operating on a globalscale, which has achieved a high level ofexcellence developed over the years. Ranking 6thin Europe and amongst the top 20 pharmaceuticalcompanies worldwide, Sanofi-Synthelabo is theflagship of French pharmaceutical research andindustry around the world.Seronosponsorwww.serono.com.auWe are one of the world's top three biotechnologycompanies focussing on four core therapeuticareas; Growth & Metabolism, Reproductive Health& Multiple Sclerosis. Serono invests more than20% of its sales in research and development.We have over 30 projects in preclinical or clinicaldevelopment, 13 new molecules and over 20discovery projects aiming to identify new drugcandidates. We aim for continuous productimprovement and the support of growthprofessionals including medical and patienteducation. We are proud of supporting the workof Australian doctors in developing the "Hormones& Me" series of booklets. One.click®, Serono'snew autoinjection device for the delivery of Saizenis designed with patient convenience in mind. It isthe only such device which injects and deliversgrowth hormone together with only one action.Servier Laboratories Site Nos: 54-57HAWTHORN VIC 3122Phone: 03 8823 7333Fax: 03 9822 4546www.servier.comServier is a privately owned Frenchpharmaceutical company that has grown fromhumble beginnings in 1955 with 9 employees, tomore than 13,000 employees today. Servier'sproducts are the result of in-house discovery anddevelopment primarily in the areas of metabolicand cardiovascular disease. 250/0 of Servier'srevenues are reinvested annually in research anddevelopment.Servier Australia's commercial interests arepresently in hypertension and heart disease(Coversyl - perindopril, Coversyl Plus ­perindopril/indapamide and Natrilix SR 1.5mg ­indapamide), diabetes (Diamicron MR -gliclazide)and disseminated malignant melanoma(Muphoran - fotemustine).1213

ESA & SRB Annual Scientific Meeting 2002ESA & SRBJoint ProgramAdelaide Convention CentreSunday, 22 September 2002toWednesday, 25 September 2002Sunday, 22 September 2002New Horizons in Reproductive BiologySymposium Conference Room 24:00pm EPIGENETIC PHENOMENA IN REPRODUCTIVE BIOLOGYMichael Holland4:30pm EPIGENETIC INHERITANCE IN MAMMALSEmma Whitelaw5:00pm SUPRAMOLECULAR SCIENCE TO NANOTECHNOLOGYLisandra Martin5:30pm GRAPES AND WINE FLAVOURPatrick WilliamsExhibition FoyersRiverview 3Eli Lilly Welcome FunctionWomen in Endocrinology Function4:00pm - 6:00pm6:00pm - 7:30pm7:00pm - 8:30pmRiverview 2ESA Council dinner and Meeting8:00pm - 11 :30prn1415

Monday, 23 September 2002Japan-Australia PlenaryPlenary Lecture Conference Hall 0 8:45am - 9:45am8:45am THE DISCOVERY OF GHRELIN - GROWTH HORMONE AND APPETITE STIMULATING HORMONEMasayasu KojimaSRB Oral Presentations - Session 1 - Male Fertility and Function 1Conference Room 108:45am - 10:00am8:30am IRON IS HIGHLY GENOTOXIC TOWARD HUMAN SPERMATOZOADennis Sawyer8:45am MORPHOLOGICAL MANIFESTATION OF Y CHROMOSOMAL MICRODELETIONS IN THE HUMANDr Marc Luetjens9:00am LEPTIN INHIBITS BASAL BUT NOT STIMULATED TESTOSTERONE PRODUCTION BY THEIMMATURE MOUSE TESTISDr Jim McFarlane9:15am THE ROLE OF FGFR SIGNALLING IN SPERM TAIL DEVELOPMENTLeanne Cotton9:30am REDOX REGULATION OF SPERM FUNCTIONMark Baker9:45am DOES CALCIUM HAS A ROLE IN SPERM HEAD ROTATION TO T-SHAPE ORIENTATION DURINGCAPACITATION? A TALE OF TWO MARSUPIAL SPECIES, THE BRUSHTAIL POSSUM (TRICHOSURUSVULPECULA) AND THE TAMMAR WALLABY (MACROPUS EUGEN/~.Dr Kuldip Sidhu..) SRB Oral Presentations - Session 2 - Embryo Development and ImplantationConference Room 28:45am - 10:15am8:30am PROLACTIN IS A PREIMPLANTATION GROWTH FACTOR IN MICEDr Peter Kaye8:45am REAL-TIME PCR EXPRESSION OF HYPOXIA-INDUCIBLE FACTORS DURING BOVINEPREIMPLANTATION EMBRYO DEVELOPMENT9:00am9:15am9:30am9:45am10:00amAlexandra HarveyEFFECT OF IN VITRO OXYGEN CONCENTRATION ON GLUCOSE TRANSPORTER-1 EXPRESSION INTHE MOUSE BLASTOCYSTKaren KindA COMPARISON OF STAINING METHODS USED TO DETERMINE NUMBERS OF CELLS IN PIGBLASTOCYSTSChristopher GrupenPARACRINE PERTURBATIONS ASSOCIATED WITH BOVINE NUCLEAR TRANSFER PREGNANCIESSusan RavelichSURVIVAL OF FRESH AND VITRIFIED SHEEP IVP AND CLONED EMBRYOS AFTER EMBRYOTRANSFERTeija PeuraABNORMAL TROPHOBLAST MHC-I EXPRESSION AND ASSOCIATED ENDOMETRIAL LYMPHOCYTICRESPONSE IN BOVINE NUCLEAR TRANSFER PREGNANCIES.Jonathan HillESA Servier Awardoral Conference Hall 0 9:45am - to:OOam9:45am SIGNALLING THROUGH THE SMAD PATHWAY BY INSULIN-LIKE GROWTH FACTOR BINDING PROTEIN­3 IN BREAST CANCERDr Susan FanayanExhibition Halls F & G16Morning Tea10:00am -10:30amESA Novartis Junior Investigator AwardConference Hall 010:30am - 12:00pm10:30am PROSTATE-SPECIFIC ANTIGEN (PSA) OVER-EXPRESSION INCREASES THE MIGRATORYPOTENTIAL OF THE PROSTATE CANCER CELLS, PC-3.10:45amTara Veveris-LoweA MOUSE MODEL OF SPINAL BULBAR MUSCULAR ATROPHY (SBMA)Patrick McNamanmy11 :OOam GENE-ENVIRONMENT INTERACTION INFLUENCES THE RELATIONSHIP BETWEEN ALCOHOLCONSUMPTION AND ABDOMINAL OBESITY IN HEALTHY FEMALE TWINS11:15amJerry GreenfieldSIGNAL TRANSDUCTION VIA 1-0-PHOSPHATIDYLINOSITOL-3 KINASE (PI3K) IS NECESSARY FORTHE SURVIVAL OF THE PREIMPLANTATION EMBRYO.Vashetharan Chandrakanthan11 :30am INHIBITION OF LIVER RECEPTOR HOMOLOGUE-1 (LRH-1) BY SMALL HETERODIMER PARTNER(SHP) IN PREADIPOCYTES.Agnes Kovacic11 :45am ROLE OF b1-INTEGRIN IN SPERMIATION AND SPERMIATION FAILUREAmanda BeardsleySRB Oral Presentations - Session 3 - Gamete Biotechnology and BreedingConference Room 1010:30am - 12:00pm10:30am CONCENTRATION OF PROGESTERONE AND SUPEROVULATORY RESPONSE WITH DIFFERENTDOSAGES OF FSHLuciene Santiago10:45am11:00am11:15amEFFECT OF ADDING SEMINAL PLASMA TO BOAR SPERM BEFORE AND AFTERCRYOPRESERVATIONRoslyn BathgateTHE INFLUENCE OF ENVIRONMENTAL TEMPERATURE ON ELITE A.1. BULLS HOUSED IN ACOOLING SHED OR FIELD AS MEASURED WITH AN INFRARED THERMOMETERAndrew BrockwayFLOW CYTOMETRIC SORTING OF FROZEN-THAWED RAM SPERMATOZOAFiona Hollinshead11 :30am EFFECT OF TIME OF INSEMINATION AND DOSE OF SORTED, CRYOPRESERVED RAM SPERM ONFERTILITY IN EWESChis Maxwell11 :45am THE EFFECT OF NUTRITION DURING THE CYCLE OF MATING ON OVIDUCTAL FLUID AMMONIAAND UREA LEVELS IN THE EWEMr Muhammad Azam KakarSRB Oral Presentations - Session 4 - Female Fertility and Function 1Conference Room 210:30am - 12:00pm10:30am PITUITARY LH~- AND FSH~-SUBUNITmRNA AND LH AND FSHCONTENT IN HEIFERS DURING ANDAFTER TREATMENT WITH GnRH AGONIST,10:45amProf Michael D'OcchioEXPRESSION OF GERM CELL AND SOMATIC CELL MARKERS DURING GONADAL DEVELOPMENTIN A MARSUPIALGratiana E Wijayanti11 :OOam LEPTIN RECEPTOR mRNA EXPRESSION DURING FOLLICULAR DEVELOPMENT AND OVULATION INTHE IMMATURE PRIMED RAT OVARYNatalie Ryan11 :15am ANDROGEN RECEPTOR EXPRESSION IN OVARIAN EPITHELIUM IN MICE OF DIFFERING AGES ANDTOTAL OVULATION NUMBER.11:30amMs Olivia TanLOCALISATION OF EPITOPES ON BRUSHTAIL POSSUM (TRICHOSURUS VULPECULA) ZONAPELLUCIDA 2 PROTEINDr Xianlan Cui11 :45am EOSINOPHILS IN DEVELOPMENT AND FUNCTION OF REPRODUCTIVE TISSUESAmanda Sferruzzi-Perri17

SRB Founders LecturePlenary Lecture Conference Hall 012:00pm EMBRYO DEVELOPMENT AND BIOTECHNOLOGYAlan TrounsonLunch (includes SRB Student Lunch Roundtable & ESA poster viewing)Exhibition Halls F & G1:OOpm - 2:00pmTransplantation and Reproduction - Issues and Solutions (SRB)Symposium Conference Room 2 2:00pm - 3:30pm~ 2:00pm DISCRIMINATING SELF AND NON-SELF: THE IMMUNOLOGICAL PARADOX POSED BY MAMMALIAN.-:;--;# REPRODUCTIONBruce Loveland2:20pm2:50pm3:10pm18PRESENT AND FUTURE OPTIONS FOR PRESERVATION OF TESTIS TISSUE AND FUNCTIONStefan SchlattDOES THE IMMUNE SYSTEM PLAY A ROLE IN REPRODUCTION? - A CLINICIANS PERSPECTIVEKelton Tremel/enFINDING SOLUTIONS: SEMEN AND MATERNAL IMMUNE TOLERANCEDr Sarah RobertsonESA Poster discussions 1 - Female Reproductionposter Exhibition Halls F & G 2:00pm - 3:15amUSE OF A HUMAN COMMERCIAL KIT TO ASSAY OESTROGEN DURING BOVINE CYCLES WITHDIFFERENT FOLLICULAR WAVESEduardo NogueiraOPTIMIZATION OF SOLID PHASE EXTRACTION PROCEDURE FOR ANALYSIS OFESTROGENS AND PROGESTOGENS IN BIOLOGICAL SAMPLESKathrin Kulhanek-~OLE OF LOCAL ESTROGEN METABOLISM IN THE CONTROL OF ESTROGEN ACTION IN HUMANPARTURITIONGemma MadsenGLUTAMATERGIC NEURONS IN THE ARCUATENENTROMEDIAL COMPLEX EXPRESS ESTROGENRECEPTOR AND PROJECT TO PREOPTIC AREA IN EWESAIda PereiraSEASONAL CHANGES IN EXPRESSION OF PREPROOREXIN (ppORX),MELANIN CONCENTRATING HORMONE (MCH) AND LEPTIN RECEPTOR (Ob-Rb) mRNA INOVARIECTOMISED (OVX) CORRIEDALE EWES IN RELATION TO APPETITE AND BREEDINGAlexandra RaoREPRODUCTIVE HORMONE RESPONSES TO ACUTE EXERCISE IN YOUNG EUMENORRHEICLeanne RedmanESTROGEN HAS A ROLE IN REGULATING APOPTOSIS IN THE BRAINRachel HillIMPAIRED MAMMARY PTHrP CONTENT AND FUNCTION AND NEWBORN GROWTH DURINGLACTATION IN THE SHRDr Mary WlodekDEVELOPING OF AN ANALYTICAL METHOD BASED ON SUPRAMOLECULAR CHEMISTRY ANDHIGH·PERFORMANCE LIQUID CHROMATOGRAPHY FOR SEPARATION AND QUANTIFICATION OFSEX STEROIDSPawel Zarzycki12:00pm -1 :OOpmESA Poster discussions 2 - HPAposter Exhibition Halls F & G 2:00pm - 3:15amTHE ACUTE ADMINISTRATION OF THE SELECTIVE SEROTONIN REUPTAKE INHIBITOR (SSRI)ANTIDEPRESSANT, SERTRALlNE, ACTIVATES THE HPA AXIS IN FEMALE, BUT NOT MALE, SHEEP.Jill Broadbear.~ THE PHYSIOLOGICAL ROLE OF GLUCOCORTICOIDS DURING EMBRYONIC DEVELOPMENT:ANALYSIS IN GLUCOCORTICOID RECEPTOR NULL MICETim ColeCORTICOTROPIN RELEASING HORMONE CAUSES VASODILATION IN HUMAN SKIN VIA MAST CELLDEPENDENT PATHWAYSRenee CromptonEFFECT OF SIZE AT BIRTH ON SALIVARY CORTISOL IN THE ADULT GUINEA PIGSanita GroverCHANGES IN ADRENAL RESPONSIVENESS DURING RAT PREGNANCY: THE ROLES OF ACTHRECEPTOR AND STAR.Susan HishehPRENATAL ENDOTOXIN EXPOSURE REDUCES STRESS RESPONSE OF PRE-WEANED GUINEAPIGS IN SEX-SPECIFIC MANNER.Nicolette TapeNEONATAL BACTERIAL ENDOTOXIN EXPOSURE ALTERS PRO-INFLAMMATORY AND FEBRILERESPONSES IN ADULT RODENTSDr Rohan Walker___);:HE EFFECT OF METYRAPONE INFUSION ON ADRENAL STEROIDOGENESIS IN THE LATEGESTATION FETAL SHEEPKirsty WarnesESA Poster discussions 3 - Inhibinposter Exhibition Halls F & G 2:00pm - 3:15amINHIBIN INCREASES MDA-MB-468 BREAST CANCER CELL ADHESION TO FIBRONECTIN.Tali LangNEOINTIMAL HYPERPLASIA IN THE RAT CAROTID INJURY MODEL IS NOT REDUCED BY LOCALFOLLISTATIN ADMINISTRATIONDavid McGrawCELLULAR RELEASE OF ACTIVIN A IN ACUTE INFLAMMATORY EVENTS IS PROSTAGLANDININDEPENDENT AND MAY REQUIRE DIRECT INTERACTION WITH LIPOPOLYSACCHARIDE (LPS).Kristian JonesEXPRESSION OF MULTIPLE TRANSFORMING GROWTH FACTOR (TGF)-~ SUPERFAMILY MEMBERSAND FOLLISTATIN IN THE ANTERIOR PITUITARYYao WangSERUM INHIBIN B & PRO-aC LEVELS WITH EXPERIMENTAL MANIPULATION OF GONADOTROPHINLEVELS IN NORMAL MEN.Dr Kati MatthiessonESA Poster discussions 4 - Metabolismposter Exhibition Halls F & G 2:04pm - 3:15pmCLONING AND EXPRESSION OF MOUSE AND RAT HOMOLOGUES OF NOVEL G PROTEIN-COUPLEDRECEPTORS, LGR7 AND LGR8; PUTATIVE RECEPTORS FOR RELAXIN AND INSULIN 3Daniel ScottFETAL GROWTH RESTRICTION IMPAIRS INSULIN SENSITIVITY OF GLUCOSE METABOLISMINDEPENDENTLY OF ADIPOSITY AND CIRCULATING FREE FATIY ACIDS IN THE ADULT GUINEADane Michael HortonAGING AND THE INSULIN RESISTANCE SYNDROME IN THE GUINEA PIG: A ROLE FOR ALTEREDBODY COMPOSITIONPrema ThavaneswaranSEPARATION OF PROSTAGLANDIN (PG)E2 AND PGF2a FROM THEIR TISSUE METABOLITESUSING REVERSED·PHASE THIN-LAYER CHROMATOGRAPHYToni WelshSOX13, HIGH SOLT, NUCLEAR LOCALISATION AND INSULIN EXPRESSIONHelen Kasimiotis19

o SITE-SPECIFICESA Poster discussions 5 - Pregnancy/Parturitionposter Exhibition Halls F & G 2:00pm - 3:15pmMATHEMATICAL MODELLING OF HUMAN MYOMETRIAL TENSION TRAJECTORIES THROUGHOUTPREGNANCYPeter SokolowskiESA Concurrent Oral Communications - Clinical 1oral Conference Room 4 3:45pm - 5:45pm3:45pm4:00pm4:15pm4:30pm4:45pm5:00pm5:15pmEXPRESSSION OF GENES IN THE UPPER AND LOWER SEGMENTS OF THE HUMANUTERUS DURING PREGNANCYKylie BrownREGULATION OF 15-HYDROXYPROSTAGLANDIN DEHYDROGENASE (PGDH) mRNA LEVELS IN THEHUMAN CHORION LAEVE AT TERM.Renee Johnson'-...QONTROL OF HUMAN PARTURITION INVOLVES CHANGES IN RESPONSIVENESS OF THE-'-"-MYOMETRIUM TO PROGESTERONE AND ESTROGENDr Sam MesianoI--.~RH GENE EXPRESSION IS REPRESSED BY ESTROGEN AND STIMULATED BY THEESTROGEN-ANTAGONIST ICI182,780 IN PLACENTARick NicholsonGonadotrophin stimulation and Risk of Multiple PregnancyProf Robert NormanGENERATION OF REACTIVE OXYGEN SPECIES BY HUMAN PLACENTAL CELLSKellie-Ann TaylorPLACENTAL RESTRICTION AND PLASMA THYROID HORMONE IN THE NEONATAL LAMB.Miles De BlasioBIOMARKER TRAJECTORIES IN PREGNANCY AND THE DETECTION OF PATHOLOGYDr Shaun McGrathDOES BULK TRANSFER OF BOVINE BLASTOCYSTS COMPROMISE EARLY EMBRYODr Jim PetersonPURIFICATION AND CHARACTERISATION OF CYTOSOLIC THYROID HORMONE BINDING PROTEININ HUMAN PLACENTABrett McKinnonExhibition Halls F & GCORTICOSTEROID BINDING GLOBULIN POLYMORPHISMS IN CHRONIC FATIGUE SYNDROMEDavid TorpyGLUCOCORTICOID RECEPTOR POLYMORPHISMS AND POST TRAUMATIC STRESS DISORDERDrAnthony BachmannPLASMA FREE, SALIVARY AND TOTAL CORTISOL RESPONSES TO VESTIBULAR CALORICSTIMULATION IN NORMAL HUMANSDr Jeff GriceDIETARY COMPOSITION IN RESTORING REPRODUCTIVE AND METABOLIC PHYSIOLOGY INOVERWEIGHT WOMEN WITH POLYCYSTIC OVARY SYNDROMELisa MoranTHE EFFECT OF INCREASED DIETARY PROTEIN ON WEIGHT LOSS AND INSULIN RESISTANCE INOBESE MEN AND WOMEN.Natalie LuscombeTHE EXTRAOCULAR MUCLE THYROID STIMULATING HORMONE RECEPTOR (TSHR) AND ITSRELEVANCE TO GRAVES' OPTHALMOPATHYSteven KloproggeMANAGEMENT OF MULTINODULAR GOITRE IN AUSTRALIA: DIFFERENCES BETWEENENDOCRINOLOGISTS AND ENDOCRINE SURGEONSJohn WalshAfternoon Tea3:30pm - 4:00pm,.,- SRB Oral Presentations - Session 5 - Female Fertility and Function 11Conference Room 24:00pm - 6:00pm5:30pm ENDOCRINE RESPONSE TO TRAUMATIC HEAD INJURY: A PROSPECTIVE STUDY.Prof Jim Stockigt20 214:00pm4:15pm4:30pm4:45pm5:00pm5:15pm5:30pm5:45pmESTRADIOL-MEDIATED CYCLIN 02 EXPRESSION: A ROLE FOR ER??DrAnn DrummondPURIFICATION OF HUMAN GRANULOSA CELLS FROM FOLLICULAR FLUID SAMPLES FOR USE INSERIAL ANALYSIS OF GENE EXPRESSION (SAGE).Michael QuinnREMODELLING OF EXTRACELLULAR MATRIX IN BOVINE FOLLICLES AT OVULATIONHelen INing-RodgersPROTEOGLYCANS - OSMOTICALLY ACTIVE MOLECULES IN FOLLICULAR FLUIDHannah ClarkeTHE INFLUCENCE OF THE OOCYTE DURING IN VITRO MATURATION ON THE METABOLISM OFBOVINE CUMULUS-OOCYTE COMPLEXESMelanie SuttonEXPRESSION WITHOUT SECRETION OF TGF-B1 AND TGF-B2 BY BOVINE AND MURINE OOCYTESLesley RitterDEVELOPMENT AND ANTRUM FORMATION IN MARSUPIAL PREANTRAL FOLLICLES CULTURED INA SERUM-FREE MEDIUMNadine RitchingsINCORPORATION OF BRDU INTO THE OVARIAN SURFACE EPITHELIUM IN MATURE MOUSE OVARYSLICES MAINTAINED IN CULTURE.Peter HurstJoint ESAlSRB (SRB 6) Concurrent Free Orals - Male Reproductionoral Conference Hall D 4:00pm - 5:45pm4:00pm4:15pm4:30pm4:45pm5:00pm5:15pm5:30pmSEX DIFFERENCES IN THE NEURONAL INPUTS FROM THE HYPOTHALAMUS AND BRAINSTEM TOTHE REGION OF THE GONADOTROPHIN-RELEASING HORMONE (GnRH) NEURONES IN THEMEDIAL PREOPTIC AREADr Chris ScottIN VITRO REGULATION OF ACTIVIN / INHIBIN SUBUNITS, FOLLISTATIN AND BAMBI IN THE 3 DAYOLD RAT TESTIS BY ACTIVIN, FOLLISTATIN AND FSHDr Kate LovelandREGULATION OF SERTOLI CELL ACTIVIN AND INHIBIN SECRETION BY HORMONES, CYTOKINESAND THE SEMINIFEROUS EPITHELIAL CYCLEMark HedgerCHARACTERISATION OF PLASMA INHIBIN FORMS IN FERTILE AND INFERTILE MENDr David RobertsonContraceptive Efficacy Of A Depot Androgen And Progestin Combination In MenProf David HandelsmanPRESENCE OF IMMUNE MODULATING MOLECULES IN BOAR SEMINAL PLASMASean O'LearyINFLUENCE OF OXYTOCIN ON OXYTOCIN RECEPTOR AND 5 a-REDUCTASE TRANSCRIPTION INTHE RAT PROSTATEMr Stephen Assinder

ESA Concurrent Oral Communications - GH/IGFConference Room 104:00pm4:15pm4:30pm4:45pm5:00pm5:15pm5:30pmMeetingMeeting4:00pm - 5:45pmTHE IDENTIFICATION AND EXPRESSION PATTERN OF A GHRELIN VARIANT THAT ENCODES ANOVEL PEPTIDE IN THE HUMAN AND MOUSEPenny JefferyOESTROGEN AND SELECTIVE OESTROGEN RECEPTOR MODULATORS EXERT DIFFERENTIALEFFECTS ON GROWTH HORMONE SIGNALLINGKin LeungINSULIN-LIKE GROWTH FACTOR-I PREVENTS APOPTOSIS IN GLUCOSE DEPLETED NEURONALCELLSDr Kisho KobayashiUROKINASE TYPE PLASMINOGEN ACTIVATOR RECEPTOR (UPAR) IS INVOLVED IN INSULIN-LIKEGROWTH FACTOR I AND II-INDUCED MIGRATION OF RHABDOMYOSARCOMA (RMS) CELLS INMarisa Gal/icchioFGF-2 PROMOTES DIFFERENTIATION OF NEUROBLASTOMA CELLS BY INHIBITION OF IGFMITOGENIC ACTIVITY AND MODULATION OF THE APOPTOTIC SIGNALING PATHWAYS.Dr Vince RussoTNFaAND IGF SYSTEM CROSS-TALK IN A HUMAN KERATINOCYTE CELL LINE: IMPLICATIONS FOREPI DERMAL HOM EOSTASIS.Dr Stephanie EdmondsonSERUM INSULIN-LIKE GROWTH FACTOR BINDING PROTEIN-5 IS RE-DISTRIBUTED IN HUMANDIABETES MELLITUS.Dr Stephen TwiggConference Hall DAdelaide University ClubAdelaide University Club22ESA Annual General MeetingSRB Student MeetingStudent BBQ5:45pm - 6:45pm7:00pm - 7:30pm7:30pm - 11 :30pmTuesday, 24 September 2002Fetal Programming (ESA)Symposium Conference Hall D 9:00am - 11 :OOam9:00am9:30am10:00am10:30amNEUROENDOCRINE AND CARDIOVASCULAR ADAPTATIONS OF THE FETUS TO ALTEREDNUTRITION- THE FETAL ORIGINS OF ADULT DISEASE?Prof Caroline McMillenPERINATAL PROGRAMMING OF SYNDROME X IN THE ADULTJulie OwensNEUROENDOCRINE PROGRAMMING OF CANCERDr Deborah HodgsonTHE PLACENTAL GLUCOCORTICOID BARRIER & FETAL PROGRAMMING: A LEPTIN CONNECTION?Dr Brendan WaddellESA Concurrent Oral Communications· Clinical 2Conference Room 119:00am9:15am9:30am9:45am10:00am10:15am10:30am10:45am9:00am - 11 :OOamTHE MORBIDITY AND MORTALITY IN GH-DEFICIENT ADULTS: EFFECTS OF LONG-TERM GHREPLACEMENT THERAPYGudmundurJohannssonUSE OF LONG ACTING SOMATOSTATIN IN ACROMEGALY: AN AUDITTien-Ming HngLONG-TERM EFFECTS OF ANDROGEN REPLACEMENT THERAPY ON BONE MINERAL DENSITY(BMD) IN MENAshrafAminorroayaACTIVATING AND INACTIVATING MUTATIONS IN THE CALCIUM SENSING RECEPTORASSOCIATED WITH DISORDERS OF CALCIUM HOMEOSTASISAaron MagnoHYPOVITAMINOSIS D - A FORGOTTEN EPIDEMIC IN THE ELDERLY WITH OSTEOPOROTICFRAGILITY FRACTURESChanna PereraSEVERE VITAMIN D DEFICIENCY IN YOUNGER ADULTS - EFFECTS ON THE SPINEDr Walter PlehweSEX-SPECIFIC CHANGES IN PLACENTAL FUNCTION AND FETAL GROWTH AND DEVELOPMENT INPREGNANCIES COMPLICATED BY ASTHMAMs Vanessa MurphyPREVALENCE AND PREDICTORS OF ANDROGEN ABUSE IN AUSTRALIAN HIGH SCHOOLSPaula AndersonSRB Oral Presentations· Session 7 • Male Fertility and Function IIoral Conference Room 1 9:00am - 11 :OOam9:00am9:15am9:30am9:45am10:00am10:15am10:30am10:45amINTROMISSION FAILURE LEADS TO INFERTILITY IN TGFB1 NULL MICEWendy IngmanIMMUNOREGULATORY CYTOKINES IN SEMINAL PLASMA OF MEN WITH INFLAMMATION ORSPERM AUTOIMMUNITYMark HedgerSEASONAL CHANGES IN THE ABUNDANCE OF 14 kDa SEMINAL PLASMA PROTEINS SHOWN TOBIND TO RAM SPERMATOZOADr John F SmithIDENTIFICATION AND LOCALISATION OF A MESOTOCIN RECEPTOR IN THE PROSTATE OF THEBRUSHTAIL POSSUM.Helen NicholsonTYROSINE PHOSPHORYLATION AND EPIDIDYMAL MATURATION IN MURINE SPERMATOZOAKelly AsquithUSE OF THE YEAST TWO-HYBRID SYSTEM TO CHARACTERISE THE FUNCTION OF SPERMPROTEIN, TPX-1Deborah HickoxGENE EXPRESSION OF ADENYLYL CYCLASE IN RAT TESTIS AND CAUDAL EPIDIDYMAL SPERMMargaret WadeSPERM SURFACE PROTEIN PH-20 EXPRESSION DURING SHEEP SPERMATOGENESISFu Yu23

ESAlSRB (SRB 8) Joint Concurrent Oral Communications - Female Reproductionoral Conference I;loom 2 9:00am - 11 :OOam9:00am OVARIAN FOLLICULAR DEVELOPMENT IS COMPROMISED IN TRANSGENIC (mREN-2)27 RATS ANDANGIOTENSIN II-INFUSED SPRAGUE DAWLEY RATSTanyth de Gooyer9:15am9:30am9:45am10:00am10:15am10:30am10:45amEFFECT OF SUB-BURSAL FAS ANTIBODY ADMINISTRATION ON FOLLICULAR CASPASE-3ACTIVATION AND OVARIAN VOLUMEMr Mark FenwickGRANULOSA CELLS MATURE BEFORE DEATH IN BASAL ATRESIADr Ray RodgersESTROGEN PROMOTES ANGIOGENESIS IN ER?-EXPRESSING HUMAN MYOMETRIALMICROVASCULAR ENDOTHELIAL CELLS BY UPREGULATING VEGF RECEPTOR-2Caroline GargettDIETARY ESTROGENS HAVE SELECTIVE EFFECTS ON THE REPRODUCTIVE AXIS IN ESTROGENDIFFICIENT ARKO MICEKara BrittA LONG TERM FOLLOW-UP OF A FEMALE WITH AROMATASE DEFICIENCYYoko MurataLIVER RECEPTOR HOMOLOGUE-1 REGULATES AROMATASE EXPRESSION IN PREADIPOCYTES.Colin ClyneMECHANISMS UNDERLYING THE REDUCED SENSITIVITY TO PROLACTIN NEGATIVE FEEDBACKDURING LACTATIONDr Stephen AndersonExhibition Halls F & GMorning TeaHarrison LecturePlenary Lecture Conference Hall DChaired by Prof. Ken Ho11 :30am ENDOCRINE-IMMUNE INTERACTIONSGeorge ChrousosExhibition Halls F & GLunch and Poster Viewing11 :OOam - 11 :30am11 :30am - 12:30pm12:30pm - 1:30pmApoptosis and Signalling (SRB)Symposium Conference Hall D 1:30pm - 3:30pm2:00pm2:10pm2:30pm3:00pm3:15pmSIGNALLING MECHANISMS DURING APOPTOSISProf Arun DharmarajanDROSOPHILA AS A MODEL SYSTEM TO STUDY STEROID HORMONE REGULATED CELL DEATHSharad KumarNEGATIVE REGULATION OF PROLACTIN RECEPTOR SIGNALLING BY SOCS PROTEINS: A NEWMECHANISM IN LUTEOLYSIS?AlProfJon CurlewisPARACRINE SIGNALLING WITHIN THE OVARIAN FOLLICLERob GilchristEXPRESSION TGFI3 SUPERFAMILY MEMBERS AND THEIR SIGNALLING COMPONENTRY BY THE RATOVARYDrAnn DrummondESA Poster discussions 6 - boneposter Exhibition Halls F & G 1:30pm - 3:00pmCYCLICAL INTRAVENOUS PAMIDRONATE IN THE TREATMENT AND PREVENTION OFOSTEOPOROSISDr Sophie ChanEXPERIENCE WITH INTRAVENOUS PAMIDRONATE THERAPY FOR THE PROPHYLAXIS ANDTREATMENT OF OSTEOPOROSIS.AlProf Duncan ToplissUP-REGULATION OF THE VITAMIN D 24-HYDROXYLASE GENE PROMOTER BY 1,25DIHYDROXYVITAMIN D: RAPID ACTIVATION AND SPECIFIC ROLES OF MAP KINASE PATHWAYS INTHE INDUCTION MECHANISM.Dr Prem DwivediESA Poster discussions 7 - cancerposter Exhibition Halls F & G 1:30pm - 3:00pmCHARACTERISATION OF A STEROID RECEPTOR-SPECIFIC TRANSCRIPTIONAL RESISTANCE IN AGRANULOSA TUMOUR CELL LINE.Simon ChuINHIBITION OF ANDROGEN SIGNALLING IN PROSTATE CANCER CELLS BY DOMINANT NEGATIVEANDROGEN RECEPTORSLisa ButlerINCREASED ANDROGEN RECEPTOR AND HER2 IMMUNOREACTIVITY IN CLINICALLY LOCALISEDPROSTATE CANCER IS ASSOCIATED WITH DISEASE PROGRESSION FOLLOWING RADICALPROSTATECTOMYDr Carmela RicciardelliLACK OF RESPONSE TO THE SYNTHETIC PROGESTIN, MEDROXYPROGESTERONE ACETATE, INADVANCED BREAST CANCER IS ASSOCIATED WITH EXPRESSION OF NON-FUNCTIONALANDROGEN RECEPTORSMiao YangESA Poster discussions 8 - clinicalposter Exhibition Halls F & G 1:30pm - 3:00pmRECOVERY OF PITUITARY FUNCTION IN XANTHOMATOUS HYPOPHYSITIS: A CASE REPORTMorton BurtINOPERABLE INTRACARDIAC PHEOCHROMOCYTOMA IN A YOUNG WOMAN WITH SUBSEQUENTCOMPLICATED BY SUBSEQUENT PREGNANCYBernard ChampionPULSED TESTOSTERONE THERAPY: PHARMACOKINETICS AND SAFETY OF INHALEDTESTOSTERONE IN POSTMENOPAUSAL WOMEN.Dr Sonia DavisonFACTORS UNDERLYING THE RISING INCIDENCE OF PAPILLARY THYROID CARCINOMA INTASMANIA (1988-1998).John BurgessFAMILIAL NEUROHYPOPHYSEAL DIABETES INSIPIDUS (FNDI) ASSOCIATED WITH A MISSENSEMUTATION ENCODING CYS58?ARG IN NEUROPHYSIN II (NPII)John Peng2425

ESA Poster discussions 9 - comparativeposter Exhibition Halls F & G 1:30pm - 3:00pmREGULATION OF RELAXIN GENE EXPRESSION IN THE CORPUS LUTEUM OF THE TAMMARWALLABY.Helen GehringSTEROID PRODUCTION BY ADRENALS AND GONADS FROM EMBRYONIC SOUTHERN SNOWSKINKS, NIVEOSCINCUS MICROLEPIDOTUS, DURING THEIR PROTRACTED GESTATION PERIOD.Dr Jane GirlingDOES HABITAT DETERMINE HORMONAL REGULATION OF METAMORPHOSIS IN AUSTRALIANANURANS?Natalie PlantLOCAL EMBRYO-DERIVED FACTORS INFLUENCE UTERINE OXYTOCIN RECEPTORS IN THETAMMAR WALLABY.Andrew SiebelTHE EFFECT OF L-CARNITINE ON ENERGY BALANCE IN THE MARSUPIAL SMINTHOPSISCRASSICAUDATA.Zoe HolthouseESA Poster discussions 10 - GH/IGFposter Exhibition Halls F & G 1:30pm - 3:00pmGROWTH HORMONE TREATMENT OF MEN WITH ABDOMINAL OBESITY REDUCES FATACCUMULATION IN THE LIVER.GudmundurJohannssonFUNCTIONAL DIFFERENCE OF SOMATOTROPES FROM AROMATASE KNOCKOUT AND WILD-TYPEMOUSE PITUITARIESDr Chen ChenEVIDENCE THAT INSULIN-LIKE GROWTH FACTOR BINDING PROTEINS HAVEINDEPENDENT AND IGF-I DEPENDENT ENDOCRINE ACTIONS IN PREGNANCYPatricia GrantMDA-MB-468 BREAST CANCER CELLS ADHESION TO FIBRONECTIN IS REGULATED BYINSULIN-LIKE GROWTH FACTOR BINDING PROTEIN-3 (IGFBP-3).Kathryn HalePRESENCE OF GROWTH HORMONE SECRETAGOGUES RECEPTOR (GHS-R) AND GHRELIN INENDOMETRIAL ADENOCARCINOMANeveen TawadrosREGULATION OF THE SOCS3 PROMOTER ACTIVITY BY GROWTH HORMONE AND OESTROGEN.Gary LeongESA Poster discussions 11 - male reproductionposter Exhibition Halls F & G 1:30pm - 3:00pmTHE MOLECULAR EFFECTS OF ANDROGENS ON SKELETAL MUSCLE CELLS IN VITROAnnaAxell26CONTROL OF SERTOLI CELL PROLIFERATION AND DIFFERENTIATIONJeremy BuzzardEXPRESSION OF 17HSD31N MOUSE PRE-ADIPOCYTES AND THE REGULATION OFANDROSTENEDIONE TO TESTOSTERONE CONVERSION IN VITRO.Lisa De CandiaTESTICULAR ACTIONS OF AN ACTIVATING MUTATION OF THE HUMAN FSH RECEPTOR INTRANSGENIC MICEAlvara GarciaDEVELOPMENT OF TRANSGENIC MICE TO EXPLORE THE FUNCTION OFPHOSPHATIDYLETHANOLAMINE BINDING PROTEINS ON MALE FERTILITYGerard GibbsA GENOME WIDE SCREEN FOR GENES ESSENTIAL TO MALE FERTILITY USING ENUClaire KennedyREDUCED N/C INTERACTIONS BUT INCREASED LIGAND INDUCED TRANSCRIPTIONAL ACTIVITY OFAN ANDROGEN RECEPTOR VARIANT CONTAINING A DISRUPTED POLYGLUTAMINE TRACTNicole MooreESA Concurrent Oral Communications - HPAConference Room 23:00pm - 5:00pm3:00pm ANALYSIS OF TRANSCRIPTION FACTORS REGULATING PLACENTAL CRH THROUGH THE CRE.Kristy Shipman3:15pm3:30pm3:45pm4:00pm4:15pm4:30pm4:45pmCORTICOSTERONE BINDING GLOBULIN EXPLOITS SERPIN MECHANISMS TO DELIVER CORTISOLTO TARGET CELLS.Dr Sinan AliREGULATION OF P450c17 mRNA BY TGF-13 SUPERFAMILY MEMBERS IN A MOUSE ADRENAL CELLLINEGuckOoiMETABOLISM OF SYNTHETIC GLUCOCORTICOIDS USED FOR THE TREATMENT OF ASTHMA BYPLACENTAL 11 ?-HYDROXYSTEROID DEHYDROGENASE TYPE 2Rebecca FittockTRANSFORMING GROWTH FACTOR B1 (TGFB1) - PUTTING THE BRAKES ON ADRENALSTEROIDOGENESIS BEFORE BIRTHDr Catherine CoulterNOVEL ASPECTS OF CRH AND AVP SIGNALLING IN PITUITARY CELLSDr Giavanna AngeliIDENTIFICATION OF AMINO ACID RESIDUES WITHIN THE TETRATRICOPEPTIDE REPEAT DOMAINOF CYCLOPHILIN 40 THAT MEDIATE HSP90 RECOGNITIONDr Bryan WardSPECIFIC LEUCINE RESIDUES WITHIN AN HSP90 C-TERMINAL MOTIF TARGETED BY THE HSP90ANTAGONIST NOVOBIOCIN ARE ESSENTIAL FOR HSP90 DIMERISATION AND INTERACTION WITHSTEROID RECEPTOR-ASSOCIATED IMMUNOPHILINSRudi AllanExhibition Halls F & GESA Concurrent Oral Communications - Male & Female Reproduction sessionSymposium Conference Hall D 3:00pm - 5:00pm3:00pm3:15pm3:30pm3:45pm4:00pmZONAL AND DEVELOPMENTAL VARIATION OF PROGESTERONE- AND GLUCOCORTICOIDRECEPTOR MRNA EXPRESSION IN THE RAT PLACENTA.Peter MarkHUMAN MYOMETRIAL GENES ARE DIFFERENTIALLY EXPRESSED IN LABOUR: A SUPPRESSIONSUBTRACTIVE HYBRIDISATION (SSH) STUDYEng-Cheng ChanINTERLEUKIN 11 SECRETION BY HUMAN ENDOMETRIAL STROMAL CELLS IS STIMULATED BYPROSTAGLANDIN E2 IN PART VIA A CYCLIC ADENOSINE MONOPHOSPHATE DEPENDENTPATHWAY.Dr Evodokia DimitriadisIDENTIFICATION OF THE TRANSCRIPTION FACTOR E2F4 AS A NOVEL BINDING PARTNER FORTHE GONADOTROPIN-RELEASING HORMONE RECEPTORAylin HanyalogluTHE INTRACELLULAR CONSEQUENCES OF THE CAG EXPANSION AND ANDROGEN RECEPTORTRUNCATION ON THE PATHOGENESIS OF KENNEDY'S DISEASEJanifer Wong4:15pm EXPRESSION AND REGULATION OF CYCLO-OXYGENASE ISOFORMS IN THE ADULT RAT TESTISUgurAIi4:30pm THE NOVEL G PROTEIN-COUPLED RECEPTOR LGR8 IS A POTENTIAL RECEPTOR FOR INSULIN 3Ross Bathgate4:45pm STEREOLOGICAL EVALUATION OF SPERMATOGENESIS INDUCED BY TESTOSTERONE (T) ORHUMAN CHORIONIC GONADOTROPHIN (hCG) IN GONADOTROPHIN DEFICIENT (hpg) MICE.Jennifer SpalivieroSRB Afternoon Tea3:00pm - 3:30pm27

ESA Concurrent Oral Communications· MetabolismSymposium Conference Room 10 3:00pm· 5:00pm3:00pm3:15pm3:30pm3:45pm4:00pm4:15pm4:30pm4:45pmDOES BETACELLULIN-INDUCED ERBB ACTIVATION INHIBIT CYTOKINE-INDUCED APOPTOSIS OFPANCREATIC b-CELLS?Natasha LloydPREDIALYSIS CHRONIC RENAL INSUFFICIENCY IS ASSOCIATED WITH ELEVATED GLUCOSELEVELS AND INSULIN RESISTANCEDr Anthony O'SuJlivanCHOLESTEROL FEEDING RESCUES THE FATTY LIVER PHENOTYPE OF THE AROMATASEKNOCKOUT (ARKO) MOUSE.Kylie HewittTHE EFFECT OF FETAL SERUM ON THE SYNTHETIC FUNCTION OF THE BRONCHIAL SMOOTHMUSCLEAnnette Osei-KumahPERIPUBERTAL INCREASES IN PLASMA LEPTIN LEVELS IN FEMALE MERINO SHEEPDr Jim McFarlanePLACENTAL RESTRICTION AND ONTOGENY OF INSULIN-REGULATED GLUCOSE HOMEOSTASIS INSHEEPDr Kathy GaffordPROGRAMMING OF APPETITE AND BODYWEIGHT BY PERINATAL DIETARY OMEGA-3 FATTYACIDS AND ANGIOTENSIN.Michael MathaiTHE EFFECT OF COMMON VARIANTS IN MTHFR ON TWINNING AND EMBRYO LOSS.Mr Grant MontgomeryESA Concurrent Oral Communications - Clinical Sex SteroidsSymposium Conference Room 11 3:00pm· 5:00pm3:00pm3:15pm3:30pm3:45pm4:00pm4:15pm4:30pm4:45pmTHE THRESHOLD FOR ANDROGEN DEFICIENCY SYMPTOMS IN MEN WITH HYPOGONADISM OFDIFFERING ETIOLOGIESMs Sharyn KelleherTHE EFFECT OF 12 MONTHS ORAL TESTOSTERONE UNDECANOATE ON BODY COMPOSITION,MUSCLE STRENGTH AND PROSTATE FUNCTION IN ELDERLY MEN.Matthew HarenTHE ACUTE EFFECTS OF HIGH DOSE TESTOSTERONE ON SLEEP IN AGEING MENDr. Peter LiuTHE RELATIONSHIP OF SERUM TOTAL TESTOSTERONE TO BODY COMPOSITION IN HEALTHYAGEING MENCarolyn AJlanTESTOSTERONE ENHANCES THE ANTINATRIURETIC AND METABOLIC EFFECTS OF GROWTHHORMONEDr James GibneyENDOGENOUS OESTROGENS BENEFICIALLY INFLUENCE ENDOTHELIAL FUNCTION IN HEALTHYYOUNG MENDr Paul KomesaroffPURIFIED ISOFLAVONES IMPROVE ARTERIAL FUNCTION IN MEN AND WOMEN.Dr Helena TeedeASSESSING THE PROGNOSTIC POTENTIAL OF CD82, A METASTASIS SUPPRESSOR, IN PRIMARYPROSTATE CANCERA/ProfAlbert FraumanSRB Junior Scientist Awards (SRB)Conference Room 13:30pm - 5:00pm3:30pm DIFFERENTIAL GENE EXPRESSION IN INTERLEUKIN-11 RECEPTOR a DEFICIENT MICE COMPAREDTO WILD TYPE DURING DECIDUALISATION3:45pm4:00pm4:15pm4:30pm4:45pmChristine WhiteMANIPULATION OF BOVINE OOCYTE-CUMULUS CELL GAP JUNCTIONAL COMMUNICATION DURINGIN VITRO MATURATION BY MODULATING CELL-SPECIFIC CAMP LEVELSRebecca ThomasLEPTIN DYNAMICS IN THE PREGNANT RAT: CHANGES IN CLEARANCE RATE ANDTRANSPLACENTAL PASSAGE LATE IN GESTATIONJeremy SmithSEMINAL CYTOKINE CONCENTRATION AND HUMAN REPRODUCTIVE OUTCOMEDavid SharkeyTHE EFFECT OF P53 DELETION ON EMBRYO VIABILITYAiqing LiINSEMINATION AND UTERINE LYMPHOCYTE TRAFFICKING IN MICEJohn BromfieldTaft Lecture (ESA)Plenary Lecture Conference Hall D 5:00pm· 6:00pm5:00pm PRE-RECEPTOR METABOLISM OF CORTISOL BY 11 ~-HYDROXYSTEROID DEHYDROGENASES:IMPLICATIONS FOR HUMANProf Paul StewartSRB Annual General MeetingMeeting Conference Room 15:00pm· 6:00pmStamford Grand, GlenelgESAISR Conference DinnerSponsored by Novo Nordisk7:00pm - 11 :30pm28 29

Thyroid Update (ESA)Symposium Conference Room 1 10:00am - 12:00pm10:00am10:30amWednesday, 25 September 2002Stem Cells (SRB)Symposium Conference Hall 0 8:30am - 9:30am8:30am9:00amGENERATING PITUITARY CELLS FROM EMBRYONIC STEM CELLS - A ROUTE TO PITUITARY CELLTHERAPYPaul ThomasTHERAPEUTIC APPLICATION OF STEM CELL TECHNOLOGIESPeter RathjenPlenaryPlenary Lecture Conference Hall B & C8:30am COMPARATIVE 3-D STRUCTURES OFTHE IGF-1 AND EGF RECEPTORSColin WardExhibition Halls F & GGrowth Factors/Cytokines (ADS/ESA)Symposium Conference Hall B & C 10:00am - 12:00pm10:00am10:30amINSULIN-LIKE GROWTH FACTORS IN DIABETES - FOR BETTER OR FOR WORSE?Prof George WertherSUPPRESSORS OF CYTOKINE SIGNALLINGDoug HiltonMorning Tea11 :OOam CONNECTIVE TISSUE GROWTH FACTORS IN DIABETIC COMPLICATIONSDr Stephen Twigg11 :30am GROWTH FACTORS OF THE VEGF FAMILY INVOLVED IN ANGIOGENESIS ANDLYMPHANGIOGENESISSteve StackerTHE ROLE OF THE TSH RECEPTOR IN THYROID ASSOCIATED OPHTHALMOPATHYA/ProfAlbert FraumanSIGNIFICANCE OF EYE MUSCLE AUTOIMMUNITY IN THYROID-ASSOCIATED OPHTHALMOPATHYJack Wall11 :OOam GENE THERAPY FOR MEDULLARY THYROID CANCERBruce Robinson11 :30amSteve Boyages8:30am - 9:30am9:30am - 10:00amHormones and Reproduction (ESA/SRB) SymposiumSymposium Conference Room 2 10:00am - 12:00pm10:00am10:30amOOCYTE-DERIVED GROWTH FACTORS AND OVULATION RATE IN MAMMALSKen McNattyINTRAOVARIAN REGULATION OF FOLLICULOGENESIS: SIGNAL MOLECULES AND THEIR CELLULARSOURCESDavid Armstrong11 :OOam OESTROGEN TARGET SITES IN HUMAN TESTISPhillipa Saunders11 :30am MALE CONTRACEPTION - APPROACH TO HORMONAL METHODSRobert McLachlanExhibition Halls F & G30Lunch and Poster Viewing12:00pm - 1:30pmPlenary Lecture1:00pm1:OOpm - 2:00pmSRB Oral Presentations - Session 9 - Models of Fertility and Infertilityoral Conference Room 1 1:OOpm - 3:00pm1:00pm1:15pm1:30pmSRB Oral Presentations - Session 10 - Uterine Biologyoral Conference Room 2 1:OOpm :' 3:00pm1:OOpm1:15pm1:30pm1:45pm2:00pm2:15pm2:30pmNeuroendocrinology of stress (with Neuroendocrinology Interest Group)Symposium Conference Room 1 2:30pm - 4:30pm2:30pm3:00pm3:30pm4:00pmFOLLICULAR DEVELOPMENT IN WALLABY OVARIAN TISSUE XENOGRAFTS IS NOT ALTERED BYTHE HORMONAL ENVIRONMENT OF THE RECIPIENTMelanie SnowDEVELOPMENT OF ANTRAL FOLLICLES IN RABBIT OVARIAN XENOGRAFTS IN IMMUNODEFICIENTMICEKarla MuirheadACUTE FSH WITHDRAWAL IMPAIRS SERTOLI AND GERM CELL DEVELOPMENT IN THE IMMATURERATDr Sarah Meachem1:45 pm EVALUATION OF TESTICULAR COOLING FOR GERM CELL TRANSPLANTATION2:00pm2:15pmZhenZhangDELIVERY OF ANTIBODIES FOR MALE IMMUNOCONTRACEPTIONAssoc Prof Russell JonesIMMUNOCONTRACEPTIVE EFFECT OF RECOMBINANT PORCINE ZONA PELLUCIDA C PROTEIN INRABBITSDr Eileen McLaughlinEFFECT OF ANGIOGENESIS INHIBITORS ON OESTROGEN MEDIATED ENDOMETRIALENDOTHELIAL CELL PROLIFERATION IN THE OVARIECTOMISED MOUSEBambang HeryantoMOLECULAR RESPONSE OF MYOMETRIAL MICROVASCULAR ENDOTHELIAL CELLS TO VEGF ANDPROGESTERONE ASSESSED BY MICROARRAYPeter RogersANALYSIS OF REPRODUCTIVE PHENOTYPE AND EXPRESSION OF CALCIUM BINDING PROTEINSIN CABP-D28K NULL MICEKien LuuMAST CELL NUMBER AND SPATIAL LOCATION IN THE VAGINAL CUL-DE-SAC OF THE BRUSHTAILPOSSUM AFTER ADMINISTRATION OF EXOGENOUS OESTRADIOLTrish MahoneyIMMUNOLOCALISATION OF INTERLEUKIN 11 AND ITS RECEPTOR IN CYCLING ENDOMETRIUM ANDIMPLANTATION SITES OF THE RHESUS MONKEY.Dr Evodokia DimitriadisREPEATED MATERNAL GLUCOCORTICOID TREATMENTS IN SHEEP IMPAIR PLACENTALSTRUCTURAL MATURATIONClaire RobertsTREATMENT WITH A GLUCOCORTICOID RECEPTOR ANTAGONIST PREVENTS BIRTH IN TAMMARWALLABIES.Geoff ShawADS/ESA Joint Plenary LectureConference Hall B & CPPARS AND THE METABOLIC SYNDROMESBart StaelsTHE NEUROENDOCRINOLOGY OF STRESSGeorge ChrousosSTRESS & STRESSORS: A CONCEPTUAL FRAMEWORKTrevor DaySTRESS & THE REPRODUCTIVE AXIS: SEX DIFFERENCES AND CONSEQUENCES FOR FERTILITYDr Alan TilbrookNEUROENDOCRINE REGULATION AND THE METABOLIC SYNDROMERoland Rosmond31

Symposium Conference Hall 8 & C2:30pm Bart Staels3:00pm Ken Kraegen3:30pm Dr John Prins4:00pm Assoc.ProfJoe ProiettoNuclear signalling (ESA/ADS)Sponsored by GlaxoSmithKline2:30pm· 4:30pmESA & SRB Annual Scientific Meeting 2002SRB PROGRAMESA! SRB Late Breaking ScienceChaired by Kate Loveland and Leon BachConference Room 23:00pm· 4:00pmAdelaide Convention Centre22-25 September 2002Sunday, 22 September 2002ISymposium - New Horizons in Reproductive BiologyChairs: Ray Rodgers and John MorrisonConference Room 24:00pm 1. EPIGENETIC PHENOMENA IN REPRODUCTIVE BIOLOGYMichael Holland4:30pm 2. EPIGENETIC INHERITANCE IN MAMMALSEmma Whitelaw5:00pm 3. SUPRAMOLECULAR SCIENCE TO NANOTECHNOLOGYLisandra Martin5:30pm 4. GRAPES AND WINE FLAVOURPatrick Williams4:00pm - 6:00pmI Eli Lilly Welcome Function I'-;E;:::X-:;::h:-;;ib:-;it;;-io---n-F=o-y--e-rs--------:!...---=------=-=-...::...:.......:..-=..::..::...:...:.::..=...:~-----6-:O-O-p-m-.-7-:3-0-p-m--;::;:~-:---::.- w__o_m_e_n_·_1n_E_nd..::..:..o.:.cr:...:.i.:..:.n..=.o.:...:lo:...::g~y~Fu=..::..:.nc=_t::io=_n:..:- 1Riverview 37:00pm· 8:30pm3233

IMonday, 23 September 2002SRB PROGRAMSRB Oral Presentations - Session 1 - Male Fertility and Function IChairs: Gerard Gibbs and Chris O'NeillConference Room 108:30am 5. IRON IS HIGHLY GENOTOXIC TOWARD HUMAN SPERMATOZOADennis Sawyer8:45am9:00am9:15am9:30am9:45am8:45am -10:00am6. MORPHOLOGICAL MANIFESTATION OF Y CHROMOSOMAL MICRODELETIONS IN THE HUMANDr Marc Luetjens7. LEPTIN INHIBITS BASAL BUT NOT STIMULATED TESTOSTERONE PRODUCTION BY THEIMMATURE MOUSE TESTISDrJim McFarlane8. THE ROLE OF FGFR SIGNALLING IN SPERM TAIL DEVELOPMENTLeanne Cotton9. REDOX REGULATION OF SPERM FUNCTIONMark Baker10. DOES CALCIUM HAS A ROLE IN SPERM HEAD ROTATION TO T-SHAPE ORIENTATION DURINGCAPACITATION? A TALE OF TWO MARSUPIALS SPECIES, THE BRUSHTAIL POSSUM (TRICHOSURUSVULPECULA) AND THE TAMMAR WALLABY (MACROPUS EUGEN/~.Dr Kuldip SidhuSRB Oral Presentations - Session 2 - Embryo Development andImplantationChairs: Eva Dimitriadis and Graham JenkinConference Room 28:30am 11. PROLACTIN IS A PREIMPLANTATION GROWTH FACTOR IN MICEDr Peter Kaye8:45am9:00am9:15am9:30am9:45am10:00am12. REAL-TIME PCR EXPRESSION OF HYPOXIA-INDUCIBLE FACTORS DURING BOVINEPREIMPLANTATION EMBRYO DEVELOPMENTAlexandra Harvey8:45am - 10:15am13. EFFECT OF IN VITRO OXYGEN CONCENTRATION ON GLUCOSE TRANSPORTER-1 EXPRESSIONIN THE MOUSE BLASTOCYSTKaren Kind14. A COMPARISON OF STAINING METHODS USED TO DETERMINE NUMBERS OF CELLS IN PIGBLASTOCYSTSChristopher Grupen15. PARACRINE PERTURBATIONS ASSOCIATED WITH BOVINE NUCLEAR TRANSFER PREGNANCIESSusan Ravelich16. SURVIVAL OF FRESH AND VITRIFIED SHEEP IVP AND CLONED EMBRYOS AFTER EMBRYOTRANSFERTeija Peura17. ABNORMAL TROPHOBLAST MHC-I EXPRESSION AND ASSOCIATED ENDOMETRIALLYMPHOCYTIC RESPONSE IN BOVINE NUCLEAR TRANSFER PREGNANCIES.Jonathan HillIMonday, 23 September 2002SRB PROGRAMSRB Oral Presentations - Session 3 - Gamete Biotechnology andBreedingChairs: Teija Peura and Graeme MartinConference Room 1010:30am -12:00pm10:30am 18. CONCENTRATION OF PROGESTERONE AND SUPEROVULATORY RESPONSE WITH DIFFERENTDOSAG ES OF FSHLuciene Santiago10:45am19. EFFECT OF ADDING SEMINAL PLASMA TO BOAR SPERM BEFORE AND AFTERCRYOPRESERVATIONRoslyn Bathgate11 :OOam 20. THE INFLUENCE OF ENVIRONMENTAL TEMPERATURE ON ELITE A.1. BULLS HOUSED IN ACOOLING SHED OR FIELD AS MEASURED WITH AN INFRARED THERMOMETERAndrew Brockway11 :15am 21. FLOW CYTOMETRIC SORTING OF FROZEN-THAWED RAM SPERMATOZOAFiona Hollinshead11 :30am 22. EFFECT OF TIME OF INSEMINATION AND DOSE OF SORTED, CRYOPRESERVED RAM SPERM ONFERTILITY IN EWESChis Maxwell11 :45am 23. THE EFFECT OF NUTRITION DURING THE CYCLE OF MATING ON OVIDUCTAL FLUID AMMONIAAND UREA LEVELS IN THE EWEMuhammad Azam KakarSRB Oral Presentations - Session 4 - Female Fertility and Function IChairs: Ann Drummond and Philippa SaundersConference Room 210:30am - 12:00pm10:30am 24. PITUITARY LH~- AND FSH~-SUBUNIT mRNA AND LH AND FSH CONTENT IN HEIFERS DURINGAND AFTER TREATMENT WITH GnRH AGONISTMichael D'Occhio10:45am25. EXPRESSION OF GERM CELL AND SOMATIC CELL MARKERS DURING GONADAL DEVELOPMENTIN A MARSUPIALGratiana E Wijayanti11 :OOam 26. LEPTIN RECEPTOR mRNA EXPRESSION DURING FOLLICULAR DEVELOPMENT AND OVULATIONIN THE IMMATURE PRIMED RAT OVARY1Natalie Ryan11:15am27. ANDROGEN RECEPTOR EXPRESSION IN OVARIAN EPITHELIUM IN MICE OF DIFFERING AGESAND TOTAL OVULATION NUMBER.Olivia Tan11 :30am 28. LOCALISATION OF EPITOPES ON BRUSHTAIL POSSUM (TRICHOSURUS VULPECULA) ZONAPELLUCIDA 2 PROTEINXianlan Cui11 :45am 29. EOSINOPHILS IN DEVELOPMENT AND FUNCTION OF REPRODUCTIVE TISSUESAmanda Sferruzzi-PerriI Morning Tea I'-;E;:::x"':';h::";ib;::-;i::;ti'":-o-=-n7 H =-a7: n- s -=F:-:&=-=G-----------:::::.-----------1-0-:0-0-a-m---1-0-:3-0-a-m-3435

IChair: John HearnConference Hall D12:00pmMonday, 23 September 2002SRB PROGRAMSRB FOUNDERS LECTUREProfessor Alan TrounsonMonash University30. EMBRYO DEVELOPMENT AND BIOTECHNOLOGY12:00pm .. 1:OOpmLunch (includes SRB Student Lunch Roundtable with Professor Trounson)Exhibition Halls F & G1:OOpm .. 2:00pmI Symposium - Transplantation and Reproduction - Issues and Solutions IChairs: Mark Hedger and Wendy IngmanConference Room 22:00pm .. 3:30pm2:00pm 31. DISCRIMINATING SELF AND NON-SELF: THE IMMUNOLOGICAL PARADOX POSED BYMAMMALIAN REPRODUCTIONBruce Loveland2:20pm 32. PRESENT AND FUTURE OPTIONS FOR PRESERVATION OF TESTIS TISSUE AND FUNCTIONStefan Schlatt2:50pm 33. DOES THE IMMUNE SYSTEM PLAY A ROLE IN REPRODUCTION? - A CLINICIANS PERSPECTIVEKelton Tremel/en3:10pm 34. FINDING SOLUTIONS: SEMEN AND MATERNAL IMMUNE TOLERANCE.Sarah Robertson1--;;::::;::7.:7.:7:::-;-::::-;;:--;:-:::-:::- A__ ft:...:,e....:rn..::..o.:..o:.,:n:...:...-=.T-=e.=.a 1Exhibition Halls F & G3:30pm .. 4:00pmISRB Oral Presentations - Session 5 - Female Fertility and Function IIChairs: Rob Gilchrist and Eileen McLaughlinConference Room 24:00pm .. 6:00pm4:00pm 35. ESTRADIOL-MEDIATED CYCLIN D2 EXPRESSION: A ROLE FOR ERa?Ann Drummond4:15pm 36. PURIFICATION OF HUMAN GRANULOSA CELLS FROM FOLLICULAR FLUID SAMPLES FOR USE INSERIAL ANALYSIS OF GENE EXPRESSION (SAGE)4:30pm4:45pmMichael Quinn37. REMODELLING OF EXTRACELLULAR MATRIX IN BOVINE FOLLICLES AT OVULATIONHelen Irving-Rodgers38. PROTEOGLYCANS - OSMOTICALLY ACTIVE MOLECULES IN FOLLICULAR FLUIDHannah ClarkecontinuedI5:00pm5:15pm5:30pm5:45pmMonday, 23 September 2002SRB PROGRAM39. THE INFLUCENCE OF THE OOCYTE DURING IN VITRO MATURATION ON THE METABOLISM OFBOVINE CUMULUS-OOCYTE COMPLEXESMelanie Sutton40. EXPRESSION WITHOUT SECRETION OF TGF-B1 AND TGF-B2 BY BOVINE AND MURINE OOCYTESLesley Ritter41. DEVELOPMENT AND ANTRUM FORMATION IN MARSUPIAL PREANTRAL FOLLICLES CULTURED IN A SERUM-FREE MEDIUMNadine Ritchings42. INCORPORATION OF BRDU INTO THE OVARIAN SURFACE EPITHELIUM IN MATURE MOUSEOVARY SLICES MAINTAINED IN CULTURE.Peter HurstJoint ESAlSRB Session 6 Oral Communications - Male ReproductionChairs: John Aitken and Peter LiuConference Hall D4:00pm .. 5:45pm4:00pm 43. SEX DIFFERENCES IN THE NEURONAL INPUTS FROM THE HYPOTHALAMUS AND BRAINSTEMTO THE REGION OF THE GONADOTROPHIN-RELEASING HORMONE (GnRH) NEURONES IN THEMEDIAL PREOPTIC AREA4:15pm4:30pm4:45pm5:00pm5:15pm5:30pmChris Scott44. IN VITRO REGULATION OF ACTIVIN / INHIBIN SUBUNITS, FOLLISTATIN AND BAMBI IN THE 3 DAYOLD RAT TESTIS BY ACTIVIN, FOLLISTATIN AND FSHKate Loveland45. REGULATION OF SERTOLI CELL ACTIVIN AND INHIBIN SECRETION BY HORMONES, CYTOKINESAND THE SEMINIFEROUS EPITHELIAL CYCLEMark Hedger46. CHARACTERISATION OF PLASMA INHIBIN FORMS IN FERTILE AND INFERTILE MENDavid Robertson47. CONTRACEPTIVE EFFICACY OF A DEPOT ANDROGEN AND PROGESTIN COMBINATION IN MENDavid Handelsman48. PRESENCE OF IMMUNE MODULATING MOLECULES IN BOAR SEMINAL PLASMASean O'Leary49. INFLUENCE OF OXYTOCIN ON OXYTOCIN RECEPTOR AND 5 a-REDUCTASE TRANSCRIPTION INTHE RAT PROSTATEStephen Assinder1 S_R_B_S_tu_d_e_n_t_M_e_e_t_in-=9:..-- ----IIAdelaide University Club7:00pm .. 7:30pmI'-- S_t_ud_e_n_tB_8_0 1Adelaide University Club7:30pm .. 11 :30pm3637

Tuesday, 24 September 2002SRB PROGRAMSRB Oral Presentations - Session 7 - Male Fertility and Function IIChairs: Sarah Meachem and Mark BakerConference Room 19:00am 50. INTROMISSION FAILURE LEADS TO INFERTILITY IN TGFB1 NULL MICEWendy Ingman9:15am9:30am9:45am10:00am10:15am10:30am10:45am9:00am - 11 :OOam51. IMMUNOREGULATORY CYTOKINES IN SEMINAL PLASMA OF MEN WITH INFLAMMATION ORSPERM AUTOIMMUNITYMark Hedger52. SEASONAL CHANGES IN THE ABUNDANCE OF 14 kDa SEMINAL PLASMA PROTEINS SHOWN TOBIND TO RAM SPERMATOZOAJohn F Smith53. IDENTIFICATION AND LOCALISATION OF A MESOTOCIN RECEPTOR IN THE PROSTATE OF THEBRUSHTAIL POSSUM.Helen Nicholson54. TYROSINE PHOSPHORYLATION AND EPIDIDYMAL MATURATION IN MURINE SPERMATOZOAKelly Asquith55. USE OF THE YEAST TWO-HYBRID SYSTEM TO CHARACTERISE THE FUNCTION OF SPERMPROTEIN, TPX-1Deborah Hickox56. GENE EXPRESSION OF ADENYLYL CYCLASE IN RAT TESTIS AND CAUDAL EPIDIDYMAL SPERMMargaret Wade57. SPERM SURFACE PROTEIN PH-20 EXPRESSION DURING SHEEP SPERMATOGENESISFuYuJoint ESAlSRB Session 8 Oral Communications - Female ReproductionChairs: Rob Norman and Jock FindlayConference Room 29:00am - 11 :OOam9:00am 58. OVARIAN FOLLICULAR DEVELOPMENT IS COMPROMISED IN TRANSGENIC (mREN-2)27 RATSAND ANGIOTENSIN II-INFUSED SPRAGUE DAWLEY RATSTanyth de Gooyer9:15am9:30am9:45am10:00am10:15am10:30am10:45am59. EFFECT OF SUB-BURSAL FAS ANTIBODY ADMINISTRATION ON FOLLICULAR CASPASE-3ACTIVATION AND OVARIAN VOLUMEMark Fenwick60. GRANULOSA CELLS MATURE BEFORE DEATH IN BASAL ATRESIARay Rodgers61. ESTROGEN PROMOTES ANGIOGENESIS IN ER?-EXPRESSING HUMAN MYOMETRIALMICROVASCULAR ENDOTHELIAL CELLS BY UPREGULATING VEGF RECEPTOR-2Caroline Gargett62. DIETARY ESTROGENS HAVE SELECTIVE EFFECTS ON THE REPRODUCTIVE AXIS IN ESTROGENDIFFICIENT ARKO MICEKara Britt63. A LONG TERM FOLLOW-UP OF A FEMALE WITH AROMATASE DEFICIENCYYoko Murata64. LIVER RECEPTOR HOMOLOGUE-1 REGULATES AROMATASE EXPRESSION IN PREADIPOCYTESColin Clyne65. MECHANISMS UNDERLYING THE REDUCED SENSITIVITY TO PROLACTIN NEGATIVE FEEDBACKDURING LACTATIONStephen AndersonTuesday, 24 September 2002SRB PROGRAMI Morning Tea ILE-X-h-i-bi-ti-o-n-H-a-II-S-F-&-G--------=.::.::.=..:.:..:.:.:..:.s!--=...::...:.::...---------:;:11;-:::~OO~a::m=--";1~1::-;;3;;-O=am=-IChair: Ken HoConference Hall DHarrison Lecture11 :30am 66. THE NEUROENDOCRINOLOGY OF STRESSGeorge Chrousos11 :30am -12:30pmI Lunch I-E-X-h-ib-i-ti-o-n-H-a-II-S-F-&-G----------...:=::...::.::.::..:..-------------::;1;-;;2;:-::3:;;O;:p=m:--~1:;-:::;;30n;p::m=-IChairs: Arun Dharmarajan and David PhillipsConference Hall D2:00pm2:10pm2:30pm3:00pm3:15pmSymposium - Apoptosis and Signalling1:30pm - 3:30pm67. SIGNALLING MECHANISMS DURING APOPTOSISArun Dharmarajan68. DROSOPHILA AS A MODEL SYSTEM TO STUDY STEROID HORMONE REGULATED CELL DEATHSharad Kumar69. NEGATIVE REGULATION OF PROLACTIN RECEPTOR SIGNALLING BY SOCS PROTEINS: A NEWMECHANISM IN LUTEOLYSIS?Jon Curlewis70. PARACRINE SIGNALLING WITHIN THE OVARIAN FOLLICLERob Gilchrist71. EXPRESSION TGF~ SUPERFAMILY MEMBERS AND THEIR SIGNALLING COMPONENTRY BY THERAT OVARYAnn DrummondI SRB Afternoon Tea ILE-X-h-i-bi-ti-o-n-H-a-II-S-F-&---=G-------=.:....:..=..:...::.::..=.~::..=.;::....:.......::-.:.....:....:...---------:;3~:On;O;:p::m:-:--;;'3:;-;j3'i\Op~m;:-3839

Tuesday, 24 September 2002SRB PROGRAMWednesday, 25 September 2002SRB PROGRAMIChair: John HearnConference Room 13:30pm3:45pm4:00pm4:15pm4:30pm4:45pmSRB Junior Scientist Awards72. DIFFERENTIAL GENE EXPRESSION IN INTERLEUKIN-11 RECEPTOR a DEFICIENT MICECOMPARED TO WILD TYPE DURING DECIDUALISATIONChristine White3:30pm· 5:00pm73. MANIPULATION OF BOVINE OOCYTE-CUMULUS CELL GAP JUNCTIONAL COMMUNICATIONDURING IN VITRO MATURATION BY MODULATING CELL-SPECIFIC CAMP LEVELSRebecca Thomas74. LEPTIN DYNAMICS IN THE PREGNANT RAT: CHANGES IN CLEARANCE RATE ANDTRANSPLACENTAL PASSAGE LATE IN GESTATIONJeremy Smith75. SEMINAL CYTOKINE CONCENTRATION AND HUMAN REPRODUCTIVE OUTCOMEDavid Sharkey76. THE EFFECT OF P53 DELETION ON EMBRYO VIABILITYAiqing Li77. INSEMINATION AND UTERINE LYMPHOCYTE TRAFFICKING IN MICEJohn BromfieldI SRB Annual General Meeting I~C:::-o-n--'f;-e-re-n-c-e-=R:-o-o-m-1::----------------_---':~:""'------5:-O-O-pm-.-s-:O-O-p-m-Stamford Grand, GlenelgESA/SRB Conference DinnerSponsored by Novo Nordisk7:00pm - 11 :30pmSymposium - Stem CellsSponsored by the CRC for Innovative Dairy ProductsChairs: Michael Holland and Craig Smith8:30am - 9:30am78. GENERATING PITUITARY CELLS FROM EMBRYONIC STEM CELLS: A ROUTE TO PITUITARY CELLTHERAPY?Paul ThomasConference Hall 08:30am9:00am79. THERAPEUTIC APPLICATION OF STEM CELL TECHNOLOGIESPeter Rathjen1~ M_o_rn_i_n..;;:;g_T_e_a :-:-__~:-:-_1Exhibition Halls F & G9:30am - 10:00amIESAlSRB Joint Symposium - Hormones and ReproductionChairs: Guck Ooi and David HandelsmanConference Room 210:00am - 12:00pm10:00am 80. OOCYTE-DERIVED GROWTH FACTORS AND OVULATION RATE IN MAMMALSKen McNatty10.30am 81. INTRAOVARIAN REGULATION OF FOLLICULOGENESIS: SIGNAL MOLECULES AND THEIRCELLULAR SOURCESDavid Armstrong11.00am 82. OESTROGEN TARGET SITES IN HUMAN TESTISPhillipa Saunders11 :30am 83. MALE CONTRACEPTION - APPROACH TO HORMONAL METHODSRobert McLachlan1~ Lu_n_ch --:-:--=-=-_:-=-:-_1Exhibition Halls F & G12:00pm - 1:30pmISRB Oral Presentations - Session 9 - Models of Fertility and InfertilityChairs: Stefan Schlatt and Jeremy ThompsonConference Room 11:OOpm - 3:00pm1:00pm 84. FOLLICULAR DEVELOPMENT IN WALLABY OVARIAN TISSUE XENOGRAFTS IS NOT ALTERED BYTHE HORMONAL ENVIRONMENT OF THE RECIPIENTMelanie Snow1:15pm 85. DEVELOPMENT OF ANTRAL FOLLICLES IN RABBIT OVARIAN XENOGRAFTS INIMMUNODEFICIENT MICEKarla Muirhead1:30pm 86. ACUTE FSH WITHDRAWAL IMPAIRS SERTOLI AND GERM CELL DEVELOPMENT IN THEIMMATURE RATSarah Meachem1:45 pm 87. EVALUATION OF TESTICULAR COOLING FOR GERM CELL TRANSPLANTATIONZhenZhang2:00pm 88. DELIVERY OF ANTIBODIES FOR MALE IMMUNOCONTRACEPTIONRussell Jones2:15pm 89. IMMUNOCONTRACEPTIVE EFFECT OF RECOMBINANT PORCINE ZONA PELLUCIDA C PROTEININ RABBITSEileen McLaughlin4041

IIWednesday, 25 September 2002SRB PROGRAMSRB Oral Presentations - Session 10 - Uterine BiologyChairs: Sarah Robertson and Anne TurnerConference Room 2 , 1:00pm - 3:00pm1:00pm 90. EFFECT OF ANGIOGENESIS INHIBITORS ON OESTROGEN MEDIATED ENDOMETRIALENDOTHELIAL CELL PROLIFERATION IN THE OVARIECTOMISED MOUSEBambang Heryanto1:15pm 91. MOLECULAR RESPONSE OF MYOMETRIAL MICROVASCULAR ENDOTHELIAL CELLS TO VEGFAND PROGESTERONE ASSESSED BY MICROARRAYPeter Rogers1:30pm 92. ANALYSIS OF REPRODUCTIVE PHENOTYPE AND EXPRESSION OF CALCIUM BINDING PROTEINSIN CABP-D28K NULL MICEKien Luu1:45pm 93. MAST CELL NUMBER AND SPATIAL LOCATION IN THE VAGINAL CUL-DE-SAC OF THE BRUSHTAILPOSSUM AFTER ADMINISTRATION OF EXOGENOUS OESTRADIOLTrish Mahoney2:00pm 94. IMMUNOLOCALISATION OF INTERLEUKIN 11 AND ITS RECEPTOR IN CYCLING ENDOMETRIUMAND IMPLANTATION SITES OFTHE RHESUS MONKEYEva Dimitriadis2:15pm 95. REPEATED MATERNAL GLUCOCORTICOID TREATMENTS IN SHEEP IMPAIR PLACENTALSTRUCTURAL MATURATIONClaire Roberts2:30pm 96. TREATMENT WITH A GLUCOCORTICOID RECEPTOR ANTAGONIST PREVENTS BIRTH IN TAMMARWALLABIESGeoff ShawChairs: Leon Bach and Kate LovelandConference Room 2ESA! SRB Late Breaking Science3:00pm - 4:00pmSociety for Reproductive BiologyThirty-third Annual Scientific MeetingABSTRACTS(Nos 1-96 in order of presentation)1. EPIGENETIC PHENOMENA IN REPRODUCTIVE BIOLOGYMichael K HollandMonash Institute ofReproduction & Development, Monash University, Clayton, Vic 3168At many levels from monozygotic twins, to standard laboratory animals, to cultures of differentiated cells, it is clear thatgenotype and phenotype are not synonymous. This is in part because of epigenetic changes. These are heritable changesin gene function involving modification in chromatin or in gene function but not structure. Epigenetic changes are seenat many points in development including development of the mammalian germ line. These modifications includedemethylation and selective remethylation of the genome, chromatin remodelling, erasure of allele specific methylationof imprinted loci and re-activation of the silent X-chromosome. Imprinting is the epigenetic modification that marks agamete's haploid genome as maternal or paternal. Imprinted genes have roles in prenatal growth, development ofparticular lineages, in behavior and in some diseases. Imprints are initiated during gametogenesis, inherited by maturegametes and transmitted to embryos. Whilst we do not as yet completely understand all the aspects of the linkagesbetween methylation, chromatin remodelling and imprinting it is clear these epigenetic phenomena function in concert ifdevelopment is to proceed normally. One example of where this may go awry is in cloning by nuclear transfer. Genes,which are silenced in somatic cell nuclei used as nuclear donors in cloning, must be reactivated for clones to developnormally. This requires reprogramming in an entirely different context than that applying during gametogenesis andwithin a much shorter time span. It is therefore not surprising that partial or total failure of reprogramming occurs. Thiswill result in a range of phenotypes in clones with only a small percentage showing complete reprogramming. Thiscorrelates with the low percentage of healthy clones. Clearly more complete understanding of epigenetic processeswould contribute substantially to our understanding of normal development and improve our of efficiency cloning. Thisis of particular importance ifhealthy cloned animals are to be produced for commercial purposes.2. EPIGENETIC INHERITANCE IN MAMMALSE. Whitelaw, V. Rakyan, J. Preis, M. Blewitt, R. Druker and S. ChongDepartment ofBiochemistry, University ofSydney, NSW 2006, AustraliaIt is well recognised that there is a surprising degree of phenotypic variation among genetically identical individualseven when the environmental influences, in the strict sense of the word, are identical. Genetic textbooks acknowledgethis fact and use different terms such as "intangible variation" or "developmental noise" to describe it. We believe thatthis intangible variation results from the stochastic establishment of epigenetic modifications to the DNA nucleotidesequence. These modifications, which may involve cytosine methylation and chromatin remodelling, result in alterationsin gene expression that, in tum, affect the phenotype of the organism. Recent evidence, from our work and that of othersin mice, suggests that these epigenetic modifications, which in the past were thought to be cleared and reset on passagethrough the germline, may sometimes be inherited to the next generation. This is termed epigenetic inheritance, andwhile this process has been well recognised in plants, the recent findings in mice force us to consider the implications ofthis type of inheritance in mammals. At this stage we do not know how extensive this phenomenon is in humans but itmay well tum out to be the explanation for some diseases which appear to be sporadic or show only weak geneticlinkage.Morgan H., Sutherland H., Martin D.I.K. and Whitelaw E. (1999) Epigenetic inheritance at the agouti locus in themouse. Nature Genetics, 23, 314-317.Whitelaw E and Martin D.I.K (2001) Retrotransposons as epigenetic mediators of phenotypic variation in mammals.Nature Genetics, 27, 361-365.Rakyan V K, Blewitt M, Droker R, Preis J, and Whitelaw E (2002) Metastable epialleles in mammals. Trends inGenetics (Vol 18, in press).4243

3. SUPRAMOLECULAR SCIENCE TO NANOTECHNOLOGYLisa MartinSchool ofChemistry, Physics and Earth Sciences, Flinders University, South AustraliaMany processes in biological and medicinal science are characterised by supramolecular chemistry. This chemistry isdefined as non-molecular with non-covalent interactions or associations between biological molecules. Examplesinclude gene regulation and transcription, protein-protein, protein-antibody interactions. Furthermore, the synthesis ofproteins or cofactors is often achieved by use of chaperone proteins assisting the "intermediates" to exist in otherwise"reactive" environments prior to delivery or assembly ofthe mature biomolecule.The most recent development in scientific technology has been the decrease in size from micro-scale to nano-scale.This has drawn into sharp focus the possible use of biological molecules as components in nano-scale devices. Thetypes of technology envisaged includes photonic devices, such as Rhodopsin, molecular motors such as, ATPase,memory chips for quantum computers based on DNA, single molecule sensitive biosensors and targeted drug deliveryand gene therapies. One of the major scientific developments that has lead to this new nanotechnology is theavailability of new materials such as microporus coatings, high tensile carbon tubes, and nanocomposite ceramics thatcan be used in bioelectronics. In addition, the development of Scanning Probe Microscopy has resulted in routineimaging at the molecular and atomic scale. This talk aims to demystify some of these components of nano-science andprovide an awareness ofpossible applications in the bio-medical sciences.Proteins and enzymes are particularly well designed to perform specific reactions with high selectivity and thereforethey are ideal biological elements for biosensors. Of particular focus in this talk is the use of supramolecular strategiestogether with immobilised techniques for metalloproteins and enzymes. These methods enable the direct, real timemonitoring of redox behaviour, which can be related to enzyme activity or function. One aim in this research is to usethe molecular properties of proteins and enzymes with high specificity and selectivity to underpin the developments ofa new generation of biosensors. The attachment of metalloproteins to an electrode surface also allows one to probe thethermodynamic and kinetic controls of their function. The methods used for enzyme immobilisation will be presentedas well as examples of the possible scope for application to other areas of biological and medicinal science.potential for new bio-medicinal diagnostics will be discussed.4. GRAPES AND WINE FLAVOURPatrick J WilliamsThe Australian Wine Research Institute, PO Box 197 Glen Osmond South Australia 5064A brief overview of research into wine and grape flavour highlighting selected developments in the field over the lastthree decades. The talk will be concerned with factors that shaped the direction and evolution of wine flavour researchrather than with details of the techniques that have been employed. For example, the focus away from wine and ontothe grape for an understanding of varietal flavours, the recognition of a multiplicity of flavour active components, andthe absence of single flavour impact compounds, and a better understanding of the origins of flavour components, arejust some of the features that define our present knowledge of wine flavour composition. Some applications of theknowledge gained from the research and the potential of this to further enhance Australia's position as a premium wineproducer will be discussed.The5. IRON IS HIGHLY GENOTOXIC TOWARD HUMAN SPERMATOZOADE Sawyer and RJ AitkenDiscipline ofBiological Sciences, School ofEnvironmental and Life Sciences, The University ofNewcastle, Callaghan,NSW, Australia, and The Hunter Medical Research Institute, NSWAustraliaHuman spermatozoa have been reported to be susceptible to the DNA damaging effects of pro-oxidants such ashydrogen peroxide (HzOz). However, DNA damage in individual genes has not been investigated in these cells. Thus,we have applied a quantitative PCR assay (QPCR) to measure gene-specific DNA damage in human spermatozoatreated with the pro-oxidants HzOz and iron. Methods: Human spermatozoa were treated in vitro with hydrogenperoxide (0-5 mM) or Fe(II)S04 (0-400 I-LM) for one hour at room temperature. After DNA purification, gene-specificDNA damage was measured by QPCR. DNA damage (e.g., strand breaks, bulky adducts) blocks the progression of thePCR polymerase, resulting in decreased amplification of the target genes. The presence of single-strand DNA breaksand alkali-labile sites was determined by alkaline gel electrophoresis. Results: Millimolar concentrations of HzOz wererequired to induce DNA damage in human spermatozoa as assessed by QPCR. After 5 mM HzOz, amplification wasreduced by 40-50%, depending on the particular gene. The nature of this DNA damage was revealed by alkaline gelelectrophoresis to be single-strand breaks and alkali-labile sites. In contrast, iron induced a significant amount of DNAdamage after treatment with as little as 100 I-LM (25-40% decreased amplification), while 400 !-LM iron reducedamplification of target genes by greater than 90%. Interestingly, the DNA damage induced by iron was not due tostrand breads or alkali-labile sites as assessed by alkaline gel electrophoresis. Discussion: These results indicate thatnuclear DNA of mature spermatozoa is relatively protected from the damaging effects of hydrogen peroxide; however,iron appears to be highly genotoxic to nuclear DNA of human spermatozoa. The two genotoxins also appear to inducedistinctly different lesions. HzOz induces strand breaks and/or alkali-labile sites, while iron does not induce theselesions. We hypothesize that iron is stimulating the release of genotoxic lipid peroxides, which then form covalentadducts with DNA bases. These results are the first to show that DNA damage can be measured in specific DNAsequences in male germ cells, and provide new insight into the susceptibility of human spermatozoa to genotoxic a~ents.6. MORPHOLOGICAL MANIFESTATION OF Y CHROMOSOMALMICRODELETIONS IN THE HUMAN TESTISeM Luetjens, M Simoni, E Gromoll and E NieschlagInstitute for Reproductive Medicine ofthe University Muenster; Domagkstr. 11; D-48129 Muenster, GermanyInfertility affects 10-15% of couples seeking offspring [1]. Between 2 - 20% of the severe cases carry Y-chromosomalmicrodeletions in the azoospermia factor (AZF) region [2]. These chromosomal microdeletions are associated withmorphological disorders of spermatogenesis. Although the missing genes are described, no principle causing germ celldepletion is known. The aim of this study is to characterise biopsies of AZF patients more intensively to find similaritiesamong the pathologies of such patients. Testicular biopsy specimens from 17 patients with. AZFc deletions wereanalysed. In a randomised study the diameter of the tubules, the lumina, the lamina propria and the epithelia weremeasured and compared with those of patients with idiopathic Sertoli-cell-Only (SeO) syndrome (n=II), mixed atrophy(n=10) and complete spermatogenesis (n=II). The biopsies were also stained for Sertoli cell-specific antigens. Additionallythe distal and proximal regions of the microdeletion were analysed with new primers in order to characterise the microdeletionposition and length [3]. By comparing the testis parameters of the AZFc deletion group with the control patients, anintermediate state between total sea and complete spermatogenesis emerged. The diameters of the average tubule of patientswith an AZFc deletion and patients with idiopathic sea were significantly smaller then those of patients with completespermatogenesis. The lumen diameter was significantly smaller compared to the control group with complete spennatogenesisand larger than in idiopathic sca. Analysis of the chromosomal microdeletions demonstrated that all patients with a deletionsolely in the AZFc-region have identical breakpoints. The immunohistochemical examination of the Sertoli cell proteins did notreveal any specific expression differences among the patient groups. AZFc microdeletions lead to alterations in testismorphology different from those in patients with mixed atrophy, although the germ cell status of both are identical. Patents withY chromosome microdeletions show an intermediate status between idiopathic seo and complete spermatogenesis. Sertoli cellfunction seems not to be altered [1] Nieschlag, E. (2000) Scope and goals of andrology. In: Nieschlag E. and Behre H. M.(eds.) Andrology: male reproductive health and dysfunction. 2nd ed. Springer, Heidelberg; 4-5.[2] Vogt et al. (1996).Human Molecular Genetics, 5, 933-943.[3] Kuroda-Kawaguchi et al. (2001) Nature Genetics, 29. 279-286.4445

7. LEPTIN INHffiITS BASAL BUT NOT STIMULATED TESTOSTERONEPRODUCTION BY THE IMMATURE MOUSE TESTISMurren Herrid and Jim McFarlaneSchool ofBiomedical,Biological and Molecular Science, University ofNew England, Armidale, NSW 2351Leptin is secreted into the blood by adipose tissue and was originally identified as a regulator of appetite and energyexpenditure. It has subsequently been shown to be involved in many other physiological systems includingreproduction. In humans and mice, plasma leptin levels are lower in males than in females even when corrected forbody fat. Some authors have attributed this difference to a suppressive effect of testosterone on leptin production.However, we have recently shown that plasma leptin levels in both intact and castrate male sheep are lower than thatfound in females, (3) suggesting that the lower levels in males are not induced by testosterone. In contrast it has alsobeen suggested that elevated leptin may suppress testosterone production, since obese men tend to have reducedfertility and reduced plasma testosterone. Recently it has been reported that leptin suppresses testosterone productionin hCG stimulated adult rat testis slices (1) and purified Leydig cells (2). In contrast, we have reported that leptin has noeffect on adult mouse Leydig cells so the aim ofthis study was to investigate the effect ofleptin on testicular slicesfrom immature (14 days old) and adult (60 days old) Swiss mouse testes. A biphasic response was observed with leptinsuppressing testosterone production by up to 60% in the unstimulated immature testis at a dose of 6.25 ng/ml. In thehCG stimulated testis leptin had no effect on testosterone production. In the adult testis leptin had no significant effecton either stimulated or unstimulated testosterone production. In contrast purified Leydig cells from both immature andadult testes were not affected by leptin. These data suggest that physiological doses of leptin do not directly affectLeydig cell steroidogenesis but acts via paracrine signaling from other cell types within the testis.(1) Tena-Sempere et. al., 1999 Journal ofEndocrinology 161: 211-218(2) Caprio et. ai., 1999 Endocrinology 140: 4939-4947(3) Kauter etal. 2000. Journal ofEndocrinology 166: 127-1358. THE ROLE OF FGFR SIGNALLING IN SPERM TAIL DEVELOPMENTLeanne M Cotton, Gerard M Gibbs, John R Morrison, Kim Sebire, David M de Kretser and Moira K O'BryanMonash Institute ofReproduction and Development, Monash University, Victoria 3168Unusual sperm tail morphology is present in a high percentage of infertile males, and because motile sperm areessential for unassisted fertilisation, research into sperm tail proteins will have an important impact on the way in whichinfertility is treated. A major focus of our group is to characterise the function of the outer dense fibres (ODF) of thesperm tail. ODF provide rigidity and directionality to tail movement, but the recent isolation of ODF proteins withenzymatic or regulatory characteristics, suggests they have a more active role in motility. Screening of a rodent testislibrary using an ODF specific antisera and 3'RACE identified SNT-2 (Suc-l associated associated Neurotrophic FactorTarget 2) as a potential ODF component. The SNTs are known FGF (Fibroblast Growth Factor) signalling adaptorsand key components in the activation of the MAP kinase and PI3-kinase pathways via the FGF receptor. SubsequentlyNorthern blot analysis revealed that SNT-2 is highly expressed in the testis and up-regulated concordant with theappearance of spermatids and sperm tail development. Further, Western blot analysis has demonstrated that rat spermtails are immunopositive for FGFR-1, and there are large concentrations of FGFs within the epididymis. Collectivelythis data suggests that FGF signalling through SNT-2 is involved in sperm tail development/function. In order to testthis hypothesis we have created transgenic mice that express a dominant-negative variant of the FGFR-1 in haploidgerm cells. The transgene is a fusion of cDNAs encoding a truncated FOFR-1 (extracellular and transmembranedomains) and a flag-tag, driven by the mouse protamine promoter. Mice have been generated by microinjection and arecurrently undergoing genotyping prior to assessing spermatogenesis and fertility.9. REDOX REGULATION OF SPERM FUNCTIONMark A. Baker, Anton Krutskikh and R. John AitkenDiscipline ofBiological Sciences, University ofNewcastle, Australia 2308.Mammalian sperm function is redox regulated. Using a variety of chemiluminescent and colorimetric probes we havediscovered multiple sources of redox activity in human spermatozoa. Thus NADH- and NADPH-dependentoxidoreductases have been detected in human sperm that generate profound chemiluminescent responses whenlucigenin is used as the probe, but not luminol, isoluminol or MCLA. The activities observed when NADH or NADPHis used as substrate are highly correlated, but biochemically distinct. For example, the NADPH-dependent activity canbe inhibited with diphenylene iodonium, while the NADH response cannot. On the other hand, both oxidoreductasescan be suppressed with the thiol reactive agent, pCMBS and both enzyme systems are competent to reduce tetrazoliumsalt derivatives such as NBT and WST-1. The NADH-dependent oxidoreductase has been isolated by FPLC and aprotein band correlating with enzyme activity was analysed by MALDI TOF. The isolated protein was identified ascytochrome bs reductase (R.2). Transient transfection of COS7 cells with this enzyme reveal an increase in NADHdependentlucigenin chemiluminescence, consistent with a redox role for cytochrome bs in human spermatozoa. TheNAD(P)H lucigenin reductase activity in human spermatozoa is significantly correlated with the induction of lucigenindependentchemiluminescent responses with 12-myristate, 13-acetate phorbol ester (PMA). Moreover, both the PMAand NAD(P)H responses are significantly elevated in populations of defective human spermatozoa recovered from thelow density region of PercoIl gradients. In contrast, the calcium-dependent chemiluminescent responses typical ofNOX5 are similar in both Percoll fractions. These observations shed light on the biochemical nature of the lesionspresent in male infertility and will help resolve the aetiology of this condition.10. DOES CALCIUM HAVE A ROLE IN SPERM HEAD ROTATION TO T-SHAPEORIENTATION DURING CAPACITATION? A TALE OF TWO MARSUPIALS SPECIES,THE BRUSHTAIL POSSUM (TRICHOSURUS VULPECULA) AND THE TAMMARWALLABY (MACROPUS EUGENIl)K.S.Sidhu, K.E. Mate and J.C.RodgerMarsupial CRC, Macquarie University, Sydney, Australia 2109Australian marsupial sperm attain T-shape orientation in the oviduct (1,2) as also during in vitro oviduct co-culturesystems (3-5). The T-shape sperm in the brushtail possum were able to bind, penetrate and fuse with eggs (6,7). Herewe purpose a hypothesis explaining the mechanism by which sperm turn T-shape during capacitation. MATERIALSAND METHODS The preparation of oviduct-conditioned media (CM) and induction of T-shape orientation in spermwere as described previously (2-6). Ionic calcium in CM was estimated using iSTAT chip. The effects of tyrosinephosphorylation and various stimulators of cAMP pathways on sperm were according to Sidhu et aI. (unpublished).RESULTS AND DISCUSSION The calcium levels in the possum oviduct is maintained at ~lmM as also in theoviductal CM produced in vitro. Estimating calcium in batches of CM indicated that the low levels of calcium areessential for optimal fertilization in the possum. Ejaculated wallaby semen contains 1.45-mM of calcium and sperm arestreamlined, unable to fertilize eggs. Resuspending the wallaby ejaculated sperm in calcium-free buffer i.e. TALP andthe possum epididymal sperm in low calcium buffer i.e. (pSOF media, calcium -0.7 mM) turned them to T-shapewithin 2 h and that was reversible with 1.45 mM calcium. Increasing cAMP or tyrosine phosphorylation in sperm alsoturned them to T-shape orientation within 1 h. In the possum, >80% sperm turn T-shape in pSOF media and thesesperm were unable to penetrate zona. Thus T-shape in sperm is not the sole criterion for capacitation. These resultsdemonstrate that low levels of calcium are essential for obtaining T-shape orientation in sperm and the latter involvescAMP and tyrosine phosphorylation pathways and are essential for fertilization. ACKNOWLEDGEMENT Thisstudy was supported by the Marsupial CRC Australia and MAF Policy New Zealand. REFERENCES IBedford, J.M.& Breed, W.O. (1994). BioI. Reprod. 50, 845-854. 2Molinia, et aI. (1998). J.Reprod. Fertil. 112, 9-17. 3Sidhu, et aI.(1998). lReprod. Fertil. 114, 55-61. 4Sidhu, et aI. (1999a). BioI. Reprod. 61, 1356-1361. 5Sidhu, et al. (1999b).Reprod. FertiI. Dev. 11, 329-336. 6Mate et aI. (2000). Zygote 8, 189- 196. 7Sidhu et aI. (2001). Proc. Aust. Soc. Reprod.BioI. 32, 22. ksidhu@possum.bio.mq.edu.au4647

11. PROLACTIN IS A PREIMPLANTATION GROWTH FACTOR IN MICEK. Markham and P.L. KayeSchool ofBiomedical Sciences, University ofQueensland, Brisbane QLD 4074Many hormones, cytokines and growth factors support optimal development of the preimplantation embryo. The closephysiological relationship (1) between growth hormone, which has already been identified as a preimplantation growthfactor (2) and prolactin, and evidence from gene knock-out models, implicated prolactin (3) and its receptor (4) inpreimplantation development. The effects of prolactin on growth of mouse embryos in vitro were thereforeinvestigated. Two-cell embryos were collected from superovulated Quackenbush mice 48 h post-hCG and incubatedwith ovine prolactin and/or antibodies against prolactin or the prolactin receptor for approximately 2 days. Grossmorphological development and the number of cells in blastocysts and their component tissues were assessed byspreading 96 h post hCG blastocysts and nuclear staining. Confocal laser scanning immunohistochemistry (CLSIH) asdescribed (2) demonstrated expression and localisation of prolactin and the prolactin receptor. Prolactin increased thetotal number of cells in the blastocyst. Concentration studies showed prolactin to be extremely potent in this role withEC50 < 1 pgfml and concentrations above 10 pgfml to be less effective. Addition of either of the blocking antibodiesreduced cell number in blastocysts developing in the absence of exogenous prolactin and blocked the effects of addedprolactin. CLSIH showed prolactin to be present in blastocysts that had been cultured 24 h. The receptors were presentin oocytes and embryos throughout the preimplantation developmental stages, coincident with mRNA expression.This suggests that embryos express prolactin receptors that are responsive to prolactin secreted in an embryonicautocrine or paracrine regulation of cell number during preimplantation development. The predominantlytrophectodermal location of prolactin receptors in the late blastocyst suggests that these receptors may be associatedwith regulation ofimplantation a major function of these cells.1. Waters MJ and Kaye PL (2002). Growth hormone & IGF research. (in press)2. Pantaleon M, et al, (1997) Proc Natl Acad Sci USA. 94(10):5125-30.3. Horseman ND, et aI, (1997) EMBO J. 16(23):6926-35.4. Ormandy CJ, et aI, 1997 Genes Dev. 11(2):167-78.Supported by NHMRC.13. EFFECT OF IN VITRO OXYGEN CONCENTRATION ON GLUCOSETRANSPORTER-1 EXPRESSION IN THE MOUSE BLASTOCYSTKaren L. Kind, Rachael A. Collett, Alexandra J. Harvey and Jeremy G. ThompsonReproductive Medicine Unit, Department ofObstetrics and Gynaecology, University ofAdelaide, The Queen ElizabethHospital, Woodville, SA, 5011.Preimplantation embryos develop in vivo under conditions of low oxygen. Oviductal and uterine oxygen concen~ationsare less than half that of atmospheric levels, with uterine O 2 concentrations of 3-5% reported around the time ofblastocyst formation in several species (1). Decreasing the concentration of oxygen during in vitro culture, to moreclosely resemble in vivo conditions, facilitates embryo development (2) and these effects maybe mediated throughaltered gene expression. Mouse preimplantation embryos express mRNA for hypoxia-inducible factor-la (3), anoxygen-regulated transcription factor which activates the expression of genes involved in the adaptation to low oxygen,including glucose transporter-l (GLUTl). The aim of this study was to determine whether oxygen concentra~onregulates expression of GLUTI in the mouse embryo. One-cell embryos were collected from CBAxC57B16 Fll111ce22 hours post hCG injection. Embryos were cultured in KSOM media under 7% O 2 ,6% CO 2 , 87% N 2 • After 48 hours,compact morulae were transferred to· 2%, 7% or 20% O 2 and cultured for a further 40 hours to the blastocyst stage.Embryos were snap frozen for RNA extraction. In vivo developed blastocysts were collected 88 hours post hCG.Extracted RNA was reverse transcribed and the abundance of GLUT1 mRNA and 18S rRNA was analysed by real-timeRT-PCR using SyBr Green Master Mix in a GeneAmp 5700 Sequence Detection System (Applied Biosystems).GLUT1 results were normalised to 18S rRNA abundance. Oxygen concentration influenced GLUTI gene expression(P

15. PARACRINE PERTURBATIONS ASSOCIATED WITH BOVINE NUCLEARTRANSFER PREGNANCIESSusan Ravelich a , Bernhard Breier a , Jim Peterson b , Jeffery Keelan a , David Wells b , Rita Lee baThe Liggins Institute, Faculty ofMedical and Health Sciences, University ofAuckland, b Reproductive Technologies,AgResearch, Ruakura Research Centre, Hamilton, New ZealandThe regulation of fetal and placental growth and differentiation involve a variety of endocrine, autocrine/and orparacrine signals, including the insulin-like growth factor family (IGFs) and leptin. Increasing evidence suggests thatparacrine signalling plays a fundamental role in placental growth and development. Alterations in the paracrine systemin the endometrium may underlie pathological processes such as infertility, preeclampsia or endometrial neoplasia(l)·Abnormal placental development and function and abnormal fetuses have been associated with in vitro derivedembryos and especially embryos generated by nuclear transfer (NT) (2). The aim of the research was to explore thehypothesis that paracrine factors involved in the regulation of normal placental growth, development and function aredysregulated in cattle pregnant with embryos generated by NT. Reproductive tracts from pregnant Friesian cows wereobtained at days 50, 100 and 150 of gestation following embryo transfer with NT or in vitro produced (IVP) embryosor following artificial insemination (AI). Placental localisation of insulin like growth factor binding proteins 2 and 3(IGFBP-2, -3) and leptin was performed by immunohistochemistry. Spatial and temporal differences in the localisationof IGFBP-2, -3 and leptin were observed in placental tissues from NT compared with AIIIVP pregnancies. At day 50and 100 of gestation, increased immunostaining for IGFBP-2 was seen in the stromal tissue below the p1acentomes inNT pregnancies: Increased staining for IGFBP-3 was observed in uterine stromal tissue in NT pregnancies at all timepoints examined. Leptin immunostaining in the fetal villi in NT pregnancies was markedly increased at day 50 and 150of gestation. In summary, the spatial and temporal patterns of expression of a number of growth factors involved inplacental and fetal growth is altered in placental tissues from NT pregnancies. It is likely that placental dysfunction inNT pregnancies is associated with paracrine perturbations via a mechanism of action as yet unknown. (1) Reis FM,Cobellis L, Luisi S, Driul L, Florio P, Faletti A, Petraglia F. (2000) Paracrine/autocrine control of female reproduction.Gynecol Endocrinol 2000 Dec;14(6):464-75 (2) Wells, D. N., Misica, P. M., and Tervit, H. R. (1999) Production ofcloned calves following nuclear transfer with cultured adult mural granulosa cells. Biology ofReproduction 60 (4), 996­1005.16. SURVIVAL OF FRESH AND VITRIFIED SHEEP IVP AND CLONED EMBRYOSAFTER EMBRYO TRANSFERTeija T. Peura, Katrina M. Hartwich, Jennifer M. Kelly, Hamish M. Hamilton, David O. Kleemann, Skye R.Rudiger, Simon K. WalkerSouth Australian Research and Development Institute, Turretfield Research Centre, Rosedale 5350, SAThe aim of this study was to examine embryo survival rates following the transfer of in vitro produced (IVP) andcloned sheep embryos cryopreserved using the Open Pulled Straw (OPS) vitrification method (1). Initially, vitrifiedDay 6 IVP blastocysts were transferred to recipients (two embryos per recipient) either within 1-2 h (=DELAYED; 34embryos) or 2 min (=DIRECT; 30 embryos) of warming. No significant differences in pregnancy or fetal survival rates(76.5 and 66.7%; 52.9 and 46.7%, respectively) were observed between the methods. Both procedures were used incloning experiments involving either serum-starved or actively growing adult granulosa and fibroblast donor cells andboth in vivo and in vitro derived oocytes. Recipient pregnancy rates and fetuses going to term for in vivo-fresh (178embryos), in vivo-OPS-DELAYED (15 embryos), in vitro-fresh (103 embryos), in vitro-OPS-DELAYED (113embryos) and in vitro-OPS-DlRECT (28 embryos) groups were 53.7,83.3,43.1,35.4 and 14.3% and 12.9,33.3, 15.5,5.8 and 0%, respectively. With in vivo derived embryos, fresh embryos yielded surprisingly significantly poorer resultsthan vitrified embryos, although there were very small numbers in the latter group. Vnlike the initial observations onIVP-embryos, the pregnancy rates achieved with in vitro-derived cloned embryos were significantly lower with OPS­DIRECT method as compared with OPS-DELAYED or fresh transfers, and fetal survival rates were significantly lowerwith both OPS-methods compared with fresh transfers. These results suggest that in vivo-derived cloned embryosendure vitrification better than in vitro-derived clones, and that the delayed transfer after OPS-vitrification is thepreferred option for somatic cell cloned sheep embryos. (1) Vajta G, Kuwayama M, Holm P, Booth P, Jacobsen H,Greve T, Callesen H. A new way to avoid cryoinjuries of mammalian ova and embryos: the OPS-vitrification. MolReprod Dev 1998; 51: 53-5817. ABNORMAL TROPHOBLAST MHC-I EXPRESSION AND ASSOCIATEDENDOMETRIAL LYMPHOCYTIC RESPONSE IN BOVINE NUCLEAR TRANSFERPREGNANCIESJonathan R. Hill l , Patricia J. Fisher 2 , Christopher J. Davies 2 and Donald H Schlafer 2Departments ofClinical Science/ and Biomedical Sciencei, Cornell University, Ithaca, New York, USA.Somatic cell nuclear transfer (NT) has produced healthy "cloned" offspring in 7 mammalian species. Few NT embryoshowever, ever reach this stage as less than 2 out of every 100 NT embryos transferred into recipient females result inviable offspring (compared to 601100 naturally fertilized embryos). In cattle, many of these losses occur around thetime of placental attachment from the fourth week of gestation. Expression of major histocompatibility complex class I(MHC-I) by trophoblast cells and distribution of endometrial T-Iymphocyte numbers were investigated as part of anassessment of the immunological status of NT pregnancies. Six 5 week old NT pregnancies were generated, all derivedfrom the same fetal cell line. All 6 NT placentas displayed trophoblast MHC-I expression. None of 8 controls (4-7weeks old) showed any MHC-I expression. Numbers of T lymphocytes (CD3 positive) were significantly higher in theendometrium of the majority of NT pregnancies compared with controls. Trophoblast MHC-I expression is nonnallysuppressed during early gestation, and the observed MHC-I expression in the cloned pregnancies is likely to haveinduced a maternal lymphocytic response that would be detrimental to maintaining viability of the NT pregnancy.18. CONCENTRATION OF PROGESTERONE AND SUPEROVULATORY RESPONSEWITH DIFFERENT DOSAGES OF FSHLuciene Lomas Santiago l ,2, Ciro Alexandre Alves Torres!, Eduardo Terra Nogueira l ,2, Antonio Bento Mancio 1and Graeme B. Martin 2IDepartment ofAnimal Science, University Federal ofVit;osa, Vifosa - Brazil.2 School ofAnilnal Biology, University ofWestern Australia, Crawley 6009.Seventeen Nelore (Bos indicus indicus) heifers were superovulated from Day 9 to 12 of the estrous cycle using twodifferent dosages of FSH (250 or 500 VI). The ojective was to measure the plasma concentration of progesteroneduring superovulation and embryo collection, and link the values to follicular dynamics and superovulatory responsethat were monitored daily by ultra-sound until the first insemination and embryo collection. Concentration ofprogesterone was analyzed every three days from Day 0 (estrous) until Day 12, at first artificial insemination and atembryo collection. Plasma was only analyzed in heifers that responded to superovulation. Heifers that displayed a lowconcentration of progesterone,. suggesting the absence of a functional corpus luteum in the beginning of thesuperovulatory protocol, didn't have satisfactory results to FSH administration. However, more viableembryos/collection were not associated with higher plasma levels of progesterone on the day of first administration ofFSH, as would be expected, but were related to individual heifer response. The plasma concentration of progesteroneon the day of insemination was similar between heifers superovulated with 250 or 500 VI ofFSH (Tab. 1). However, onthe day of embryo collection (d=7), the levels were inversely correlated to FSH dosage and directly related to thenumber of corpora lutea following superovulation (11.25 x 8.25 CL/donor). We conclude that superovulation with 250VI of FSH was more efficient for stimulating more responsiveness follicles and the appearance of more corpora lutea;as a result, the secretion of progesterone was higher. Associated with this, all animals superovulated with 250 VI ofFSH responded to the superovulatory protocol.Table 1 - Plasma concentration of progesterone in Nelore heifers on the day of artificial insemination and the day ofembryo collection.Progesterone(ng/mL)Cycle day 250 VI FSH 500 VI FSHd=O (Artificial InsemmatlOn . .) 051 .'aA 0.35 a ,Ad-7 (Embryos collection) 41.29 a ,B 17.40 b ,BAverages followed by different lower case letter in same row are different (p

19. EFFECT OF SEMINAL PLASMA ON FROZEN·THAWED BOAR SPERMR Bathgate, BM Eriksson, WMC Maxwell and G EvansFaculty ofVeterinary Science, The University ofSydney, NSW 2006Under optimal conditions, fertility after insemination with frozen boar spenn can be close to that achieved with chilledsemen but, in the field, the levels are often substantially lower. We investigated the addition of seminal plasma (SP),reported to have beneficial effects in other species, before and after freeze-thawing, with the aim of improving postthawquality of spenn. The sperm-rich fraction of 3 ejaculates from each of 4 boars were collected and frozen,packaged in 0.5ml straws, according to the method of Bwanga et a1. (1). The experimental groups had cooling extendersupplemented with 100, 50 or 0% SP (standard protocol). After thawing, the samples were diluted inBTS containing0, 5, 10, 20, 40, 80 or 100% SP. Post-thaw addition of SP in the range of 5-40% had a beneficial effect on initialmotility and after 3h incubation at 37°C (p

23. THE EFFECT OF NUTRITION DURING THE CYCLE OF MATING ONOVIDUCTAL FLUID AMMONIA AND UREA LEVELS IN THE EWEM. A. Kakar\ S. Maddocks\ M. F. Lorimer 2 , D. o. Kleemann 3 , S. R. Rudiger 3 and S. K. Walker 3lDepartment ofAnimal Science, Adelaide University Roseworthy Campus, Roseworthy, SA 5371;2Biometrics SA, Adelaide University, Private Mail Bag1.GlenOsmond.SA 5064;3South Australian Research and Development Institute, Turretfield Research Centre, Rosedale, SA 5350This study examined the effect of changes in nutrition during the cycle of mating in the ewe on the concentrations ofammonia and urea in oviductal fluid. Twelve mature ewes (4-5 years, 58-67 kg) of comparable body condition werefed rations that provided either 1.5 x daily energy needs for maintenance (H) or 0.5 x maintenance needs (L).Nutritional treatments were imposed from 18 d before until 6 d after the expected time of ovulation. Oviducts werecatheterised for four days before the start of the experiment and oviduct fluid was collected from day 0 until day 6 afterovulation. Half of the ewes were treated to superovulate (S) (using progestagen, FSH, GnRH treatment) while theremainder (NS) ovulated spontaneously following progestagen treatment. The data were analysed by ANOVA as a 2x2factorial design. Body weight was significantly (P

27. ANDROGEN RECEPTOR EXPRESSION IN OVARIAN EPITHELIUM IN MICE OFDIFFERING AGES AND TOTAL OVULATION NUMBER.Olivia L. Tan, Peter R. Hurst, Jean S. FlemingDepartment ofAnatomy & Structural Biology, Otago School ofMedical Sciences, PO Box 913, Dunedin, New Zealand.One hypothesis of ovarian surface epithelial (OSE) carcinogenesis is that high levels of progesterone are protective,whereas high levels of androgen increase the risk of ovarian cancer (1). Epithelial-lined inclusion cysts are common inall women and are thought to be precursors of ovarian cancer (2). This study aimed to investigate spatial differences inexpression of androgen receptor (AR) in the OSE and inclusion cysts in mice of varying ages and total lifetimeovulation number (total OV#).Ovaries from Swiss Webster mice (total OV# median: range [n mice]) were collected from 3 month virgins (3 m;113: 11-235 [55]), from 12 month old breeders (12 m; 217: 97-386 [21]) and from 8 month virgin mice, housed in splitcages alongside a male, to induce continuous oestrous cycles (8 m; 629: 456-908 [16]). Ovaries were also collectedfrom prepubertal mice (4 w; 0[9]) (3). AR immunohistochemistry was performed on 4 /lm sections, cut fromparaformaldehyde-fixed, paraffin-embedded tissue. Invaginations were defined as where OSE descended into theovarian stroma, but remained connected to the OSE, while inclusion cysts were enclosed structures within the ovary,lined with epithelium. Inclusion cysts were observed only in 8 m and 12 m animals.AR expression was low in OSE from 3 m and 12 m animals, with very few squamous cells staining. However in 4w and 8 m animals there were very few positive cuboidal cells also. In contrast the majority of cells lining inclusioncysts, with the exception of attenuated cells, were stained for AR. Low levels of AR expression were seen ininvaginated OSE in 12 m animals. No differences were seen between invaginated epithelium and OSE in 3 m or 8 manimals. The strong upregulation of AR expression in cyst epithelial cells suggests that androgen responsiveness mayplaya role in the aetiology of ovarian cancer.1. Risch HA (1998) Journal National Cancer Institute, 90, issue 23, 1774-17862. Blaustein A (1981) Pathology Annual, part 1,247-2493. Clow OL et al (2002) Molecular and Cellular Endocrinology 191(1), 105-11129. EOSINOPHILS IN DEVELOPMENT AND FUNCTION OF REPRODUCTIVETISSUESAmanda Sferruzzi-Perri, Ann Hallett, Sarah Robertson, Lindsay DentDepartments ofMolecular Biosciences and Obstetrics and Gynaecology University ofAdelaide, Adelaide SA 5000Eosinophils are a distinctive feature of some mucosal tissues, where they may be important in defence againstinfectious agents, but may also have a role in normal physiology by contributing to development and maintenance ofepithelial integrity. Several recent studies have indicated that deficiencies in eosinophils or defects in eosinophilrecruitment are associated with delayed postnatal mammary gland morphogenesis [1] and perturbations in the estrouscycle [2], [3]. These reports led us to examine the affect of an over-abundance of eosinophils on these processes. Asinterleukin 5 (IL-5) is central to the regulation of eosinophil growth and differentiation, eosinophilic IL-5 transgenic(IL-5 Tg) mice and normal wild type (WT) mice were compared to determine the impact of eosinophils on mammarygland development and reproductive function. Eosinophils were increased 13-fold and 4-fold in the uterus andmammary gland, respectively in IL-5 Tg mice compared to WT littermates. High numbers of eosinophils were alsonoted in the regressing corpora lutea of ovaries in IL-5 Tg mice whereas eosinophils were rarely seen in WT ovaries.Mammary gland development was transiently delayed in IL-5 Tg mice, with significantly less ductal extension andbranching at 7 weeks of age. IL-5 Tg mice had significantly shorter estrous cycles than WT mice (4.7±0.4 days versus6.4±0.5 days, respectively, p = 0.03). At day 18 of pregnancy, fetal resorptions are increased by 75% in IL-5 Tgheterozygote matings, and a decrease in fetal: placental ratio was observed (mean + SEM =7.54 ± 0.09 in IL-5 Tgmatings versus 8.26 ± 0.10 for WT, p < 0.001). Parturition was not compromised in IL-5 Tg mice, as judged by thegestation length and pregnancy outcomes. The birth weights and growth trajectories of pups were not significantlyinfluenced by maternal IL-5 transgene expression. Our studies suggest that IL-5 transgenesis and/or eosinophilia mayalter the estrous cycle and at least transiently delay ductal morphogenesis in the murine mammary gland. Eosinophilsrecruited into the ovary of IL-5 Tg mice may impact indirectly on both of these parameters. Despite these observations,the outcomes of pregnancy, parturition and lactation were not compromised by maternal over-expression of IL-5.References: [1] Development, 2000. 127(11): p. 2269-82. [2] J Reprod Fertil, 2000. 120(2): p. 423-32. [3]Endocrinology, 2001. 142(10): p. 4515-21.28. LOCALISATION OF EPITOPES ON BRUSHTAIL POSSUM (TRICHOSURUSVULPECULA) ZONA PELLUCIDA 2 PROTEINXianlan. Cui*, Janine Duckworth*, Susie Scobie*, Denise Jones*, and Phil Cowan tLandcare Research, *PO Box 69, Lincoln 8152, New Zealand; tprivate Bag 11052, Palmerston North, New ZealandImmunisation of female possums (Trichosurus vulpecula) with recombinant possum zona pellucida 2 constructs (rZP2)led to 72-75% reduction in fertility, presumably through interference in secondary sperm-egg binding. In order todefine those regions of the protein crucial to sperm-egg binding, antigenic regions on possum ZP2 protein weremapped. The amino acid sequence of full-length possum ZP2 protein was used to synthesise a complete set ofoverlapped 15-mer peptides. The peptides were used in a modified enzyme-linked immunosorbent assay (ELISA) toidentify continuous epitopes recognized by antibodies in the sera of possums immunised with rZP2 constructs. Thirteenepitopes were located on possum ZP2 protein. Comparisons of the ELISA binding patterns of peptides to antibodies inthe individual sera with the fertility status ofthe immunised possums revealed three significant peptide epitopes. One ofthe three epitopes bound to fixed possum spermatozoa from the caudal epididymis. Another corresponded to a similarregion that has been shown to be integral to fertilisation in the mouse. The significance of these three individualepitopes in relation to fertilisation in the possum is currently under investigation.5657

SRB FOUNDERS' LECTURE30. EMBRYO DEVELOPMENT AND BIOTECHNOLOGYAlan TrounsonCentre for Early Human Development, Institute ofReproduction and Develop11'lent and National Biotehnology CentreofExcellence - Centre for Stem Cells and Tissue Repair, Monash University, Clayton, Vic. 3168Mammalian preimplantation embryos are the only stage of development that may be grown in vitro independent of thematernal environment. The grown oocyte formed in the maturing follicle can be extracted from the ovary and eithermatured in vitro, or recovered after maturation in vivo, to produce viable embryos with a high developmentalcompetence. This has enabled the in vitro production of mammalian embryos that may be fertilized and grown to theblastocyst stage in culture. While control of the germ cell, primary oocyte and growing oocyte in the preantral follicleis less well understood, it is possible to manipulate ovarian follicular dynamics by ovarian transplantation and may becontrolled in the future by interference in the normal selection mechanisms of primordial follicle recruitment. Data isbeing generated from expression libraries in the human and other species to identify the key regulatory molecules forfollicular and oocyte recruitment. This may be used to alter the reproductive life-span for the female.The production of oocytes and embryos provides the basis for reproductive biotechnologies in domestic animal speciesand the human. Cattle reproductive technologies are well developed for production and human IVF is the foundationfor the treatment of human infertility. The cattle industry accepts embryo production, embryo cryopreservation andembryo transfer as an integral part of the breeding strategies used. More recently, the cloning of animals, particularlycattle, has been explored with variable outcomes. The reprogramming of somatic cells by ooplasm doesn't completelyreset the epigenetic regulators of gene expression, so that failure to erase the somatic epigenetic signature can result inabnormalities of development, including both placental and fetal compartments. This results in high rates ofimplantation failure, fetal growth and both anatomical and functional abnormalities which are often lethal. However,around 50% of offspring are viable and healthy despite the altered epigenetics. In the second generation the epigeneticirregularities are corrected during gametogenesis.The ability to clone animals enables the production of transgenic animals in one generation. Other methods oftransgenesis in animals include sperm-mediated transfection, which involves intracytoplasmic sperm injection (ICSI).Other new and interesting methods are being designed that involve the manipulation of gametogenesis.Human IVF has evolved into a successful treatment for female and male infertility. IeSI may used for testicular spermand elongated spermatids. However, success with round spermatids or their precursors has not been clinically useful.The new and rapidly growing area of genetic diagnosis of point mutations and trinucleotide repeat disease inpreimplantation embryos is an important new biotechnology in human medicine. These new molecular techniques arealso being applied for aneuploid screening of IVF embryos, reducing the need for multiple embryo transfer formaintaining high pregnancy success rates. It is now possible to fingerprint sibling embryos to test new culture systemsetc. With these capabilities, it is possible to identify genes that correlate with breast cancer and HLA type embryos forcompatible transplants. Expression screening and apoptosis markers may also be used for determining embryo viability.Preimplantation embryos are also a source of pluripotential embryonic stem (ES) cells. These may be derived fromfertilized oocytes, parthenogenetic embryos and nuclear transfer embryos. It is of interest that embryos that have littleor no developmental competence can produce ES cell lines with apparently complete ability to produce all types ofterminal tissue types and to integrate in vivo into all tissues of interest. This may be similar to the ability of some adultcell types to transdifferentiate under experimental conditions in vitro and in vivo. These extraordinary cells may enablea new medical biotechnology of cell therapy for tissue repair and regeneration. These cells may also allow for genetherapy to be applied safely for the treatment of a wide range of genetic diseases.These have been exciting times for biological sciences and promise a wonderful future research opportunity for youngscientists. I thank all the scientists who have worked with me over the decades for the joy it has brought me for thecontributions they have' all made. I also acknowledge the funding of our creativity by numerous agencies andcommercial bodies over the decades.31. THE IMMUNOLOGICAL PARADOX OF PREGNANCY - SURVIVAL OF THEULTIMATE ALLOGRAFTBruce E. LovelandThe Austin Research Institute, Heidelberg, VIC 3084A successful pregnancy requires that the mother accepts a foreign body. This involves paradoxes for the immunesystem. The extensive genetic diversity between mother and offspring provides protein differences that, givenexposure to the immune system, range from weakly to very strongly immunogenic. Thus the placenta and foetusrepresent "the ultimate allograft". The highly active metabolic exchange across the placenta and dynamicdevelopmental changes provide an opportunity for strong maternal immunity to develop. Establishment andmaintenance of pregnancy involves immune tolerance, immunosuppression and immunomodulation. A state ofselective maternal-anti-foetal immunosuppression exists which would be the dream of organ transplanters to mimic.Global immunosuppression obviously would be detrimental to the mother and a strong local anti-microbial immunitymust be maintained.The mammalian immune system has evolved to cohabitat with microbes and parasites. It is perceived to consist ofdifferent but overlapping compartments, described as the Innate, Humoral and Cellular. These provide, respectively,extremely rapid containment and destruction of infecting organisms, an induced mechanism to recognise and removeparticulate matter, and an induced mechanism to recognise and destroy infected cells in the body. These are achievedthrough quite distinct "pattern recognition" processes. It is apparent that selective control of each of these is crucial tosuccessful pregnancy. A central feature of the immune system is the need to distinguish "self' from "foreign" and theprovision of a variety of mechanisms to achieve this at very subtle levels. Its critical function to screen out mutating,potentially cancerous cells, is also relevant to our consideration of pregnancy and immunity. So how is the specificsuppression induced?There are several mechanisms thought to be crucial to the maternal / foetal immune state - perhaps more to beuncovered. Blocking the function of indoleamine 2,3-dioxygenase (IDO), a widely expressed enzyme responsible fortryptophan metabolism, results in maternal-anti-foetal immune responses. IDO is currently the most fashionablecandidate to explain maternal immunosuppression. Secondly, control of the complement cascade, central to the innateimmune system, is also essential to healthy pregnancy.This presentation will introduce the immune mechanisms enabling the ultimate allograft, a foetus, to survive and thrive.32. PRESENT AND FUTURE OPTIONS FOR PRESERVATION OF TESTIS TISSUE ANDFUNCTIONStefan SchlattInstitute ofReproductive Medicine, University Munster, Domagkstr. 11, 48129 Munster, GermanyThe testis releases two major products: male gametes and male sex steroids. While absent or insufficient secretion ofsteroids can easily be treated by administration of exogenous hormones, no cure has been established for a defect ingerm cell development. The introduction of ICSI, however, has drastically changed the prospects for treatment, asgeneration of even a single postmeiotic male germ cell offers a chance of achieving a pregnancy. Therefore, newapproaches based on male germ cell transplantation and testicular tissue grafting might be optimised to work for thegeneration of progeny. Interspecies transfer of germ cells showed that the recolonisation by stem cells is robust, asspermatogonial stem cells from phylogenetically distant species can recolonise the mouse testis. In contrast to the factthat spermatogonial stem cells are able to recolonise the seminiferous tubules, complete spermatogenesis is onlyobserved when germ cells are transplanted between phylogenetically closely related species. We have developed anapproach to infuse germ cells into monkey and human testes and tested whether germ cell transplantation can beapplied as a tool for gonadal protection in non-human primates. However, due to the risk of tumour cell contaminationgerm cell transplantation will not eliminate the cryopreservation of sperm as a widely used tool and well acceptedoption for gonadal protection in postpubertal oncological patients. It appears likely that optimised techniques willinstead become available to generate male gametes from preserved testicular material. One of the promising newapproaches, and an alternative to germ cell transplantation, is grafting of testicular tissue. We have shown that testiculartissue from various newborn species grafted ectopically into the backskin of immunodeficient mice is capable ofdeveloping qualitatively complete spermatogenesis. Sperm obtained from these grafts have been used for the generationof progeny. Like the more advanced status of ovarian tissue grafting as a tool to protect fertility and substitutehormones in female patients, grafting of testicular tissue might become an important tool for fertility preservation andhormonal replacement therapy in the male.5859

33. DOES THE IMMUNE SYSTEM PLAYA ROLE IN REPRODUCTION?A CLINICIANS PERSPECTIVEKelton TremellenReproductive Medicine Unit, Adelaide, South Australia.Human reproduction is an enigma from an immunological perspective since the immune system can be responsible forinfertility and pregnancy complications, while facilitating a successful reproductive outcome in the majority. In thispresentation, a brief overview of the immune systems role in each stage of human reproduction will be presented, alongwith an example of the clinical consequences when things go wrong.Cytokines are involved in the control of all aspects of ovarian function. including follicle growth, steroid productionand ovulation. Around the time of ovulation a surge in local production of TNF-a, IL-I ~ and GM-CSF results in therecruitment of macrophages into the follicle, which inturn release proteases and prostaglandins producing rupture of thedominant follicle and ovulation. Inhibition of these processes with non-steroidal anti-inflammatory drugs has beenshown to inhibit ovulation and potentially result in infertility.The presence of allogenic sperm in the female reproductive tract provides an immunological challenge to the female. Inmost cases, repeated exposure of a woman to sperm antigens will tollerise her towards her partners antigens, whichinturn will enhance placental invasion and decrease her chances of developing pre-eclampsia. Alternatively, adversematernal immune responses to sperm can lead to infertility. Examples include the ability of anti-sperm antibodies toblock fertilisation or destruction of sperm by peritoneal macrophages activated by cytokines liberated fromendometriosis.The interaction between the embryo, endometrium and immune system has been extensively studied in rodents.Cytokines such as II-I, IL-ll, LIF, GM-CSF and CSF seem to benefit embryo development and implantation, whileIFNy and TNFa have a negative influence. While these cytokines may mediate their action directly on the embryoitself, many mediate their influence indirectly by modifying uterine leukocyte numbers and activity. Womenexperiencing recurrent miscarriage of immune aetiology have been identified as having abnormal endometrial cytokineexpression and Natural Killer cell activity.The immune system continues to playa role in human reproductive processes long after conception and earlyembryonic development. Inappropriate activation of the maternal immune system can lead to pre-term labour, fetalgrowth restriction and pre-eclampsia. Evidence will also be presented linking adverse maternal immune responses andcerebral palsy.34. FINDING SOLUTIONS: SEMEN AND MATERNAL IMMUNE TOLERANCESarah A. RobertsonDepartment ofObstetrics and Gynaecology and Reproductive Medicine Unit, The University ofAdelaide, SA 5005.Successful pregnancy requires a state of maternal immune 'tolerance' to accommodate antigens expressed by theconceptus. Implantation failure and placental pathologies such as recurrent miscarriage and pre-eclampsia largelyreflect insufficiencies in maternal immune adaptation, but progress in devising therapeutic strategies to treat theseconditions is stalled because the mechanisms underlying induction and maintenance of maternal tolerance are unknown.Increasingly, clinical and experimental data support the proposal that insemination has consequences for thereproductive process beyond delivery of male gametes. An emerging hypothesis is that insemination comprises the'inductive phase' ofthe maternal immune response to pregnancy, since it is in the context of semen that the female tractis first and most frequently exposed to paternal transplantation antigens shared by the conceptus. Our studies in miceindicate (1) that at insemination, the maternal tract is better poised to process antigens than at any other stage of thecycle or pregnancy; (2) that semen contains high concentrations of 'immune-deviating' cytokines, includingtransforming growth factor (TGF)~ and prostaglandin E, known to induce immune tolerance in other mucosae; (3) thatexposure to semen is sufficient to induce a transient state of functional immune tolerance to paternal transplantationantigens, and (4) that insemination is causally linked to the activation and expansion of populations of lymphocytesrecruited into the implantation site. Recent studies in women show that comparable events occur in the human cervixafter intercourse. Together, these studies indicate that exposure to semen may promote the success of pregnancythrough orchestrating a maternal immune response conducive to optimal placental invasion and function. Since it isthe inductive phase that principally determines the strength and quality of any immune response and is most amenableto therapeutic manipulation, we expect that unravelling the cellular and molecular processes linking insemination withimplantation will facilitate development ofnew clinical interventions for treatment of immune-based fertility disorders.35. ESTRADIOL-MEDIATED CYCLIN D2 EXPRESSION: A ROLE FOR ERa?Ann E. Drummond, Mitzi Dyson and Jock K. FindlayPrince Henry's Institute ofMedical Research, PO Box 5152 Clayton, Victoria 3168 AustraliaCyclin D2, a cell cycle regulator, plays a key role in the proliferation of ovarian granulosa cells (1) and thus follicledevelopment. Follicle stimulating hormone (FSH) and estradiol (E2) have been shown to enhance cyelin D2 mRNAexpression in vivo (2). It is unclear however, which estrogen receptor (ER), ERa. or ER~, mediates this response. Weaddressed this question using cultured granulosa cells (3), which exclusively express cyclin D2 (no other cyclins).Granulosa cells were isolated from the ovaries of diethylstilboestrol (DES)-treated, 21 day old Sprague-Dawley rats.Cells in triplicate wells, were plated overnight (day 1), the media was changed (day 2) and treatments were added onday 3 for 2 h. FSH (100ng/ml) and E2 (1 and IOnM) were added separately and in combination. RNA purified fromfreshly isolated and cultured granulosa cells, was reverse transcribed and subjected to real time PCR analysis of cyclinD2, ERa, ER~ and GAPDH (data normalisation) mRNAs. Experiments were repeated three times. Cyclin D2 mRNAexpression by freshly isolated granulosa cells was elevated 4-fold in comparison to control cells plated for 2 days. Inresponse to FSH, cyclin D2 mRNA expression was stimulated 2-fold (compared to control cells). In contrast, E2 at 1and IOnM, failed to influence cyclin D2 expression. The combined FSH and E2 treatment gave results consistent withthe stimulation of cyclin D2 mRNA by FSH alone. ER~ mRNA expression by granulosa cells did not change with timein culture or in response to treatment. ERa mRNA expression by granulosa cells, down-regulated by in vivo DEStreatment (3), was not restored with time in culture. We hypothesise that ERa not ER~ is involved in the regulation ofcyclin D2 expression by E2. The down-regulation of ERa mRNA, induced by the in vivo exposure to DES and itscontinued decline in vitro, may account for the inability of E2 to enhance cyclin D2 mRNA expression. These findingsare consistent with our belief that ERs play different roles in the ovary: ER~ mediates differentiated functions and ERacellular proliferation. [1] Sicinski et aI., (1996) Nature 384: 470-474. [2]Robker et aI., (1998) Molec. Endocrinol.12:924-939. [3]Drummond et aI., Molec. Cell. EndocrinoI. 149: 153-161. Supported by the NH&MRC of Australia(Regkey 983212).36. PURIFICATION OF HUMAN GRANULOSA CELLS FROM FOLLICULAR FLUIDSAMPLES FOR USE IN SERIAL ANALYSIS OF GENE EXPRESSION (SAGE)Michael C .I Quinn!, J L Stanton!, P A Hessian 2 , D P L Green!IMolecular Embryology Group, Department ofAnatomy & Structural Biology, University ofOtago, New Zealand2Leukocyte Inflammation Research Laboratory, Department ofPhysiology University ofOtago, New ZealandFollicular fluid collected at the time of IVF is a prime source of human granulosa cells. However, follicular fluid oftencontains red and white blood cells. This contamination must be removed for meaningful in vitro or molecular studies.Previous studies have purified granulosa cells by a combination of density gradients (1), the use of magnetic anti-CD45beads (2) and also by flow cytometry (3). We propose a purification methodology which exploits clumps of muralgranulosa cells present in follicular fluid. Follicular fluid samples were collected from female patients undergoing IVFfor male factor infertility at the Otago Fertility Center, Healthcare Otago. Centrifugation through a 50% Percollgradient (600g) was used to remove red blood cells. Cells were collected from the Percoll interface and viewed under adissecting microscope. Clumps of granulosa cells were mechanically removed with a glass pipette and washed in HamsFlO media. To determine white blood cell content cells were stained with antibodies (CD3, CD14, CD16, CD19) andquantified by flow cytometry. White blood cell contamination was found to be between 2 and 4%. An FSH antibodyconfirmed purified cells were positive for FSH. In addition, cell clumps were examined by histology and electronmicroscopy and found to contain homogenous cells, which were identified as granulosa cells. This method provides arelatively simple and fast method to purify human granulosa cells, which is applicable to molecular studies.(1) Best, C. L., Pudney, J., Anderson, D. J., Hill, J. A. 1994. Modulation of human granulosa cell steroid production invitro by tumor necrosis factor alpha: implications of white blood cells in culture. Obstetrics & Gynecology, 84, 121­127. (2) Choi, D., Lee, E, Y., Yoon, S., Hwang, S., Yoon, B, & Lee, J. 2000. Clinical correlation of cyclin D2 mRNAexpression in human luteinized granulosa cells. Journal ofAssisted Reproduction and Genetics, 17, 574-579. (3) DeNeubourg, D., Robins, A., Fishel, S., & Gibbon, L. 1996. Flow cytometric analysis of granulosa cells from follicularfluid after follicular stimulation. Human Reproduction, 11,2211-2214.6061

37. REMODELLING OF EXTRACELLULAR MATRIX IN BOVINE FOLLICLES ATOVULATIONHelen F. Irving-Rodgers!, Michael J. D'Occhio 2 , William J. Aspden 2 , Raymond J. Rodgers l1Reproductive Medicine Unit, Department ofObstetrics and Gynaecology, University ofAdelaide and 2Animal Sciencesand Production Group, Central Queensland University, Rockhampton.Within the membrana granulosa we have identified a unique extracellular matrix (fociemix) consisting of basal laminalike material that develops as follicles enlarge (l). To elucidate its role we examined its fate following ovulation, byimmunohistochemical analysis of the composition of fociemix and the follicular basal lamina. Ovulation in Bos indicuscattle (426-562 kg) was synchronized as follows: Day 1; intra-vaginal progesterone releasing device (CIDR) plus 2 mg17~-oestradiol (Cidirol); Day 9; prostaglandin (Estrumate); Day 10; removal of CIDR. Animals were. randomlyallocated to 3 groups: Group 1, Control (n =6), no further treatment; Group 2 (n =8), injection of 25mg LH (Lutropin)Lm. on Day 11; Group 3 (n = 8), 25 mg i.m. LH on Day 10. Ovaries were collected on Day 12, such that Group 2ovaries were collected 12 - 14 h and Group 3 38 - 40.5 h after LH. Pre- or recently-ovulatory follicles were dissectedfrom the ovary, in an apical-basal orientation, and frozen in OCT compound or fixed in 2.5% glutaraldehyde.Immunohistochemistry was performed on OCT embedded tissue and the following basal lamina components localised:collagen type IV (a1), laminin chains ~1, ~2, yl, nidogen, perlecan and the extracellular matrix proteoglycan versican.Group 1 follicles were 13-20 mm in diameter and Group 2 follicles 6-18 mm. Pre-ovulatory follicles in Group 3measured 10, 11 and 18 mm, and other follicles had ovulated. Following ovulation the follicular basal lamina remainedimmunopositive for laminin chains ~2, yl and nidogen. Laminin ~1 was not detected. Staining for the a1 chain ofcollagen type IV was reduced compared with basal lamina staining of smaller antral follicles (2), and discontinuous inrecently ovulated follicles, indicating degradation of the basal lamina. Perlecan was absent from the membranagranulosa and follicular basal lamina of most Group 3 pre- and post-ovulatory follicles as was versican. However,granulosa cells (especially Group 2 follicles) showed intense cytoplasmic staining for versican, suggesting it was upregulated. The findings indicate that matrix in the basal lamina and within the membrana granulosa of follicles activelychanges at the time of ovulation. (1)Biol Reprod 2001: 64, Suppl 1, Abstract 101. (2)Biol Reprod 1998: 59, 1334­1341.39. THE INFLUCENCE OF THE OOCYTE DURING IN VITRO MATURATION ON THEMETABOLISM OF BOVINE CUMULUS-OOCYTE COMPLEXESMelanie Sutton!, Pablo Cetica 2 , Robert Gilchrist!, Jeremy Thompson l1 The Reproductive Medicine Unit, Department ofObstetrics and Gynaecology, University ofAdelaide, The QueenElizabeth Hospital, Woodville, South Australia.2 Department ofBiochemistry, School ofVeterinary Science, University ofBuenos Aires, Argentina.Interactions between the oocyte and surrounding cumulus cells are bi-directional and oocyte-secreted factors haveprofound effects on cell growth and differentiation. The aims of this study were to determine the influence of theoocyte on bovine cumulus cell metabolism during in vitro maturation. Cumulus-oocyte complexes (COC) wereaspirated from antral follicles from abattoir-derived ovaries. Complexes were oocytectomised (OOX), leaving thecumulus and zona pellucida intact. Intact COC, OOX and OOX plus denuded oocytes (OOX+DO) were culturedindividually in 10JlI drops ofIVM media (TCM 199 + pyruvate + BSA + LH + FSH) at 39°C in 5% CO 2 in air. Oxygenconsumption measurements were taken at 1-4h, 11-14h and 21-24h of culture using a non-invasive microfluorescencemethod [1]. Spent media was collected and analysed for glucose and pyruvate consumption and L-lactate production[2]. The DNA content of individual complexes was quantified using PicoGreen fluorescence dye to enable metabolicmeasurements to be expressed per ng of DNA. There were no significant differences between COC, OOX andOOX+DO in any of the metabolic parameters measured, providing further evidence that oocyte-specific factors, whileregulating cell proliferation, have few measurable effects on bovine cumulus mass expansion [3]. However, regardlessof treatment, metabolic profiles changed over the 24h maturation period. There were significant increases (P500 kDa, indicating that large molecular weightmolecules contribute significantly to the overall osmotic potential of follicular fluid. Pools were then treated withenzymes (DNAse, proteinase K, hyaluronidase, chondroitinase ABC, keratanase, collagenase, or heparinase) and thendialysed at 100 kDa cut off at 37C in 2M NaCI to prevent aggregation of macromolecules, followed by dialysis at 37°Cagainst H 2 0. Osmotic pressure was reduced 42 ± 2% and 33 ± 3% by digestion of hyaluronic acid (0.5u/201l1hyaluronidase from Streptomyces hyalurolyticus, 4 h, 37°C, pH 6) in healthy and atretic follicles, repectively. Digestionof GAG chains (0.02u/20IlI, chondroitinase ABC from Proteus vulgaris, 4 h, 37°C, pH 8) resulted in a reduction in OPof 54 ± 14% and 20 ± 3.6% in fluid from healthy and atretic follicles. This suggests that PGs in follicular fluid, toolarge to readily cross the follicular wall, can exert osmotic pressure.accumulation of follicular fluid.These PGs may therefore contribute to the40. EXPRESSION WITHOUT SECRETION OF TGF-PI AND TGF·p2 BY BOVINE ANDMURINE OOCYTESLesley J Ritter, Megan P Morrissey, David T Armstrong and Robert B GilchristReproductive Medicine Unit, Department ofObstetrics and Gynaecology, The University ofAdelaide,TQEH, Woodville, South Australia.It is now well established that oocytes are potent regulators of cumulus cell function and thereby their ownmicroenvironment. This activity is commonly attributed to members of the TGF-~ superfamily because of the capacityof these growth factors to mimic oocyte-secreted factors (1). The aims of this study were to examine in bovine andmurine oocytes (1) the expression pattern of TGF-~l and ~2 mRNA throughout meiotic maturation, and (2) whetheroocytes secrete TGF-~1 and ~2. Follicular cells and oocytes were collected from abattoir derived bovine ovaries andfrom eCG-primed immature mice. RNA was extracted from follicular cells and from oocytes at defined stages ofmeiotic maturation; in mouse from both in vivo and in vitro matured oocytes at 0, 3, 6, 9 and 24h, and in cows fromimmature and in vitro matured oocytes. RNA was reverse transcribed and cDNA was amplified with specific primerstargeted against TGF-~l and ~2. Both bovine and murine denuded oocytes (DO) expressed TGF-~l and ~2 mRNA atall stages of meiotic maturation. All somatic cells examined expressed mRNA for both TGF-~ transcripts. Thepossibility of CC contamination of DO .samples was excluded by screening for FSH receptor· mRNA expression byPCR. In order to determine whether DO secrete TGF-~ proteins, mural granulosa cells (MGC) from both species werecultured in the presence of either DO or TGF-~ +/- a TGF-~ pan specific neutralising antibody. MGC were assessed for[3H]-thymidine incorporation as a marker of DNA synthesis. eH] counts were increased significantly (P

T If41. DEVELOPMENT AND ANTRUM FORMATION IN MARSUPIAL PREANTRALFOLLICLES CULTURED IN A SERUM-FREE MEDIUMN M Richings, P D Temple-Smith & M B RenfreeDepartment ofZoology, University ofMelboume, Vic, 3010Culture of isolated preantral follicles is potentially an important source of mature eggs for research and assistedreproductive techniques, where an understanding of both the prepubertal and adult system is important. In the onlypublished report of follicle culture in a marsupial species [1], the culture system supported follicular growth but noantral development was observed. The aim of this study was to compare the growth and development of preantralfollicles from prepubertal and adult tammar wallabies, Macropus eugenii, in a serum-free culture system. Preantralfollicles were dissected from ovaries of 6 prepubertal and 5 adult tammars and cultured individually in drops ofmedium (Dulbecco's modified essential medium with various additives) at 37°C in a humidified gas environment of 6%CO 2in air for 4 days. Follicles were measured and observed each day. Growth rates were expressed in terms ofproportional increase in volume over the culture period and the follicles were assessed for signs of antrumdevelopment. Almost all follicles increased in volume (Prepubertal, 53/54; Adult, 54/55) and there was no significantdifference in the average volume increase between groups (Prepubertal, 48%; Adult, 59%; p>0.05). There was,however, a significant difference between groups in the incidence of antrum formation (Prepubertal, 10/54; Adult, 0/55;p1000 OSE cells were found to be BrdU positive after injections into the animals. In culture,however, the OSE showed increasing BrdU incorporation with 5%, 20% and over 50% labelled cells at 24,48 and 72hr. respectively. No other tissues within the ovary were found to have significant numbers of labelled cells. In a fewexamples the mesothelial lining of the ligament suspending either the ovary or oviduct also had labelled cells. Theseresults indicate the considerable potential for proliferation of OSE cells in vitro. BrdU incorporation may be induced byan OSE-specific proliferative factor, or factors present in the culture medium. Alternatively the removal of the ovaryfrom the mouse may eliminate an inhibitory influence or be a wound-response to the act of dissection of the ovary.Whatever the reason, it remains to be determined if the cells of the OSE can proceed in vitro to enter and completemitotic division.44. IN VITRO REGULATION OF ACTIVIN / INIDBIN SUBUNITS, FOLLISTATIN ANDBAMBI IN THE 3 DAY OLD RAT TESTIS BY ACTIVIN, FOLLISTATIN AND FSHlTerri Meehan, 2Ann Drummond, IDavid de Kretser and lKate Loveland1Monash Institute ofReproduction and Development, Monash University, and 2 Prince Henry's Institute ofMedicalResearch, Clayton, Victoria 3147Members of the transforming growth factor (TGF)P superfamily and their antagonists are produced by several celltesticular types during early postnatal development. We have observed a discrete switch from activin to follistatinexpression within gonocytes as they transform into spermatogonia, and we have illustrated their functional effect onsomatic and germ cells (Meehan et al 2000. Dev BioI 220:225). This lead to the hypothesis that these proteins mayregulate their own expression and thereby modulate this stage of testicular differentiation. Using 24 hour cultures of 3day old rat testis fragments and real-time PCR analysis, expression levels of activin and inhibin subunits (a, pA andPB), follistatin and BAMBI mRNAs were measured in response to activin A, follistatin and FSH relative to mediaalone. BAMBI is a pseudoreceptor that antagonises signalling by activin, TGFpand some bone morphogenetic proteins,while both follistatin and inhibin can inhibit activin signalling. The inhibin a subunit rnRNA was increased 3-fold byFSH (to 280% of control, p

45. REGULATION OF SERTOLI CELL ACTIVIN AND INHffiIN SECRETION BYHORMONES, CYTOKINES AND THE SEMINIFEROUS EPITHELIAL CYCLEYoshinori Okuma, Anne O'Connor, Julie Muir, David Phillips, David de Kretser and Mark HedgerCentre for Molecular Reproduction and Endocrinology, Monash Institute ofReproduction and Development, Clayton,VictoriaActivin A and the inflammatory cytokines, interleukin-la (IL-la) and IL-6, are produced by the Sertoli cell and areimplicated in control of spermatogonial and spermatocyte development. Activin A also inhibits IL-l and IL-6production and action in several other systems, and therefore is a potential negative regulator of testicularinflammation. However, there have been few studies describing the production and regulation of activin A in the testis.Sertoli cells from immature (20 day-old) or adult rats were cultured for 48h in the presence of IL-la, IL-l~, IL-lreceptor antagonist (IL-lra), IL-6, the inflammatory mediator lipopolysaccharide (LPS from E. coli), testosteroneand/or ovine FSH or dibutyryl cAMP (cAMP). Adult rat seminiferous tubules were stage-dissected and incubated withtest substances for 72h. Dimeric activin A and inhibin B in the culture media were measured by specific ELISAs. Inimmature and adult Sertoli cells, activin A secretion was stimulated by IL-l, and inhibited by FSH/cAMP. Conversely,inhibin B was stimulated by FSH/cAMP and inhibited by IL-l. LPS induced a large increase in activin A, which waspartially inhibited by IL-lra, and a decrease in inhibin B secretion by the Sertoli cells. In cultured seminiferous tubules,activin A secretion occurred across all stages, with a distinct peak of secretion at stage VIII. This secretion was almostcompletely blocked by IL-lra. In contrast to isolated Sertoli cells, activin A secretion by cultured seminiferous tubulesat stages VII-VIII was stimulated by cAMP. Inhibin B secretion was stimulated in all stages by cAMP, but not by IL-l.These data confrrm that Sertoli cell activin A production is positively regulated by endogenous and exogenous IL-l viaa cAMP-independent pathway, although other locally-produced cytokines also may be involved. Activin A isnegatively regulated by FSH/cAMP, but this arm of the regulation is modulated by specific germ cell associations.Inhibin B, on the other hand, is positively regulated by FSH/cAMP, and negatively regulated by IL-l. There isreciprocal control of activin A and inhibin B throughout the cycle of the seminiferous epithelium by IL-l and FSH,which has important implications for control of spermatogonial development and the effects of inflammation on thepituitary-testicular axis.47. CONTRACEPTIVE EFFICACY OF A DEPOT ANDROGEN AND PROGESTINCOMBINATION IN MENL Turner, S Wishart, AJ Conway, PY Liu, E Forbes, RI McLachlan, D.I HandelsmanDepartment ofAndrology, Concord Hospital, ANZAC Research Institute, Sydney NSW 2139 & Prince Henry's InstituteofMedical Research, Monash Medical Centre, Clayton, Victoria 3168Hormonal male contraception was theoretically feasible for decades with formal proof of concept demonstrated inlandmark WHO male contraceptive efficacy studies in early 1990s. Combined testosterone plus progestin treatment isthe most promising strategy, but no contraceptive efficacy studies are reported. The superior efficacy plus lesserdemands on compliance suggest depot regimens are ideal for hormonal male contraception. We completed the firststudy estimating the contraceptive efficacy of a depot hormonal regimen, an androgen/progestin combination, in men.Fifty-five men in stable relationships seeking a change in contraceptive method were administered testosterone (four200 mg implants, 4 or 6 monthly) and a progestin (300 mg DMPA, 3 months). Once sperm output was suppressed«IM/mL, two consecutive months), men entered a 12 month efficacy period when all other contraception was ceased.Sperm output was monitored monthly. There were no pregnancies in 426 person-months (35.5 person-years) of efficacyexposure (upper 95% CL for contraceptive failure rate, 8% per annum). Mean sperm density fell rapidly (by 88% @1month and 98% @2 months) allowing nearly all men to enter efficacy within 3 months (50% @ 1 month, 83% @ 2months, 94% @ 3 months). Only 2/55 (3.6%) men were unable to enter efficacy due to insufficient suppression ofsperm output. A few men treated with T implants at 6 month intervals demonstrated androgen deficiency symptoms&/or escape of spermatogenic suppression (predicted by suboptimal gonadotropin suppression) between months 5-6;men receiving T implants at 4 months intervals had no androgen deficiency nor loss of gonadotropin and sperm outputsuppression. Recovery was slower than anticipated (median time 6-9 months to sperm density 20M/mL) presumablydue to DMPA kinetics. Discontinuations were for protocol (12), personal (10) and medical (5) reasons but there wereno serious adverse effects related to drug exposure. We conclude that the first prototype depot androgen/progestincombination regimen provides high contraceptive efficacy with satisfactory short-term safety but slow recovery ofspermatogenesis. Larger studies with purpose-developed depot combinations are required to clarify the overall safetyand efficacy of the most promising approach to hormonal male contraception. This study was supported by CONRAD.46. CHARACTERISATION OF PLASMA INHmIN FORMS IN FERTILE ANDINFERTILE MEND.M. Robertson, T. Stephenson, R.I. McLachlanPrince Henry's Institute ofMedical Research, P.O. Box 5152, Clayton, 3168, VictoriaInhibin B is the major feedback regulator of FSH in the male. In vitro data suggest that precursor high mol wt forms ofinhibin have reduced bioactivity. The aim of this study was to determine the levels of high mol wt forms of inhibin Band inhibin a-subunit in plasma in fertile men and men with various forms of infertility. Plasma from control fertilemen (n=l1), and men classified by testicular histology as hypospermatogenesis (HypoSG, n=4), Sertoli Cell Only(n=4), germ cell arrest (n=2) and Klinefelter's Syndrome (n=3), and following chemotherapy (n=4) were fractionatedusing a combined immunoaffinity chromatography, preparative SDS-PAGE and electroelution procedure. Inhibinforms were determined by ELISAs for inhibin B, total inhibin (all forms of inhibin containing the aC fragment) andpro-aC. In fertile men and men with HypoSG, inhibin B was identified as mature (26-30k) and precursor (60k) formswith similar proportions (29.1 % vs 25.2%, respectively) of the 60k form in both groups. Inhibin B levels were too lowto be assessed in the other infertile groups. Precursor forms of pro-aC (46k, pro-aN-aC) also showed no differencesbetween control (7.4%) and all infertile groups (7.4-11 %).To· establish if the Pro-aC ELISA was detecting the precursor forms of inhibin B containing the pro- fragment, normalmale plasma was repeatedly immunoabsorbed with antiserum (INPRO) to the pro- region and the remainingimmunoabsorbed plasma then immunoabsorbed with antiserum to the aC subunit. No inhibin B was detected in theINPRO-absorbed sample and the profile of immunoactivity as determined by Pro-aC and total inhibin ELISAs weresimilar. Similarly, the profiles of inhibin B and total inhibin in the a subunit-absorbed sample were identical. Thesedata indicate that the pro-aC ELISA is detecting all forms of the monomeric aC subunit but not the precursor inhibinB forms and that the inhibin B ELISA is detecting all dimeric inhibin B. The two ELISAs are thus separately detectingall the free a subunit and inhibin B in male plasma.It is concluded that a) the Pro-aC and inhibin B ELISAs provide an unambiguous assessment of the levels of theseinhibin forms in the circulation and b) the proportion of precursor inhibin B forms in plasma (25-30%) is unchanged inmen with sub-fertility.48. PRESENCE OF IMMUNE MODULATING MOLECULESIN BOAR SEMINALPLASMAS. O'Leary, D.T. Armstrong and S.A. RobertsonDept. OfObstetrics and Gynaecology, The University ofAdelaide and Reproductive Medicine Unit, The QueenElizabeth Hospital Woodville. South Australia.Factors in seminal plasma are recognised as potential modulators of reproductive success in the pig. Experiments inrodents and more recently in pigs indicate roles for specific proteins in seminal plasma in inducing cellular andmolecular changes in the female reproductive tract. An active constituent of murine seminal plasma has been identifiedas the potent immune modulating cytokine transforming growth factor beta (TGF~)l. In vitro experiments suggest theresponse to TGF~ is further influenced by interferon (IFN)y and lipoplysaccharide (LPS). To investigate whetherimmune modulating molecules are present in pig semen, the content of TGF~}, TGF~2, IFNy and LPS in boar seminalplasma were measured. Semen was collected from 42 Large White boars of similar age and of known fertility, andseminal plasma was prepared by centrifugation. Commercial ELISA assays were used to measure TGF~}, TGF~2 (bothPromega) and IFNy (Endogen) and LPS was measured by Limulus Amebocyte Lysate assay (Bio Whittaker). TGF~land TGF~2 were detected in all samples [median (range) = 185 (91-423) and 50.1 (26-101) ng/ml respectively], IFNywas detectable in only 2 samples (39 & 40 pg/ml) and endotoxin was present in 30 samples [125 (11-193) EU/ml].Variation over time and effect of frequency of collection on TGF~ content was also evaluated. Content of both isoformsvaried

49. INFLUENCE OF OXYTOCIN ON OXYTOCIN RECEPTOR AND 5 a-REDUCTASETRANSCRIPTION IN THE RAT PROSTATESteve Assinder, Mei Sun, Helen NicholsonArgo, Department ofAnatomy and Structural Biology, University ofOtago, New Zealand.Dihydrotestosterone (DRT) is essential for normal function and growth of the prostate. DRT is produced by thereduction of testosterone, catalysed by 5 a-reductase. Two isoforms of the 5 a-reductase enzyme exist. In the ratprostate type I is expressed in the epithelium and type II in stromal cells. Oxytocin increases the activity of thisenzyme but it is unclear how. Neither expression nor distribution of oxytocin receptor have previously beendescribed in the rat prostate. This study aimed to determine the distribution of oxytocin receptor, and to investigateif oxytocin influences expression of its receptor and the isoforms of 5 a-reductase. Adult male Wistar rats weretreated daily with either 5 ~glKg oxytocin, 2 ~glKg of a specific oxytocin antagonist (OTA), both oxytocin andOTA, or saline (control) respectively for 3 days. On day 4 rats were euthanased by CO2 inhalation and the ventralprostate removed. Western blot and immunohistochemistry, employing a polyclonal antibody to the 3 rd intracellularloop of the oxytocin receptor, identified a specific peptide of -60 kDa and localised oxytocin receptor to both thestromal tissue and epithelium. Total RNA was used in a semi-quantitative RT-PCR approach to estimate relativelevels of expression of 5 a- reductase type I and II, and oxytocin receptor with levels normalised against a B-actinamplicon as internal control. Oxytocin treatment significantly decreased (P

---------------------------------------------------~-~---~-~--53. IDENTIFICATION AND LOCALISATION OF A MESOTOCIN RECEPTOR IN THEPROSTATE OF THE BRUSHTAIL POSSUMHelen Nicholson!, Jo Fink!, Steve Assinderl, Bernie McLeod 2 , Laura ParrlIDepartment ofAnatomy & Structural Biology, University ofOtago, New Zealand, 2AgResearch, Invermay AgriculturalCentre, 3New Zealand, Howard Florey Institute, Melbourne.Oxytocin is present in the prostate of the eutherian mammals and has been implicated in the regulation of prostatecontractility and growth [1]. The marsupial produces mesotocin, rather than oxytocin, and a mesotocin receptor hasbeen identified in the Tammar wallaby prostate [2]. The aim of this study was to identify a mesotocin receptor withinthe possum prostate and investigate the localisation of the receptor protein in the adult prostate in and out of thebreeding season. Groups of possums (n=6-10), trapped alive in the wild, were killed at two monthly intervals duringthe year and prostate tissue collected. The prostates were divided into cranial and caudal portions and either frozen orfixed in 10% formalin. Immunohistochemistry and western blot analysis were performed using an antibody (020) raisedto the third intracellular loop of the sheep oxytocin receptor [3]. RT-PCR of total prostatic RNA using primersdesigned to the Tammar mesotocin receptor generated a 337bp amplicon. On sequencing this demonstrated. 92 %sequence homology with the Tammar receptor. Western blot analysis confirmed the presence of a single -60 kDaprotein. Immuno-reactive receptor protein was identified in all of the prostates examined and was localisedpredominantly to the epithelial cells of the glandular acini of both the cranial and caudal regions of the prostate. Littlereceptor was detected in the stromal tissue surrounding the glands. Significant changes (P

57. SPERM SURFACE PROTEIN PH-20 EXPRESSION DURING SHEEPSPERMATOGENESISFu Yu, Helen D Nicholson, Robin McDonald!, Stuart A Meyers 2 , Grant W Montgomery3, John F Smith!, Jean SFlemingDept Anatomy & Structural Biology, University ojOtago School ofMedical Sciences, Dunedin, New Zealand;IAgResearch, Ruakura Research Centre, Hamilton, New Zealand; 2Dept Anatomy, Physiology & Cell Biology,University ofCalifornia, Davis, USA; 3Queensland Inst Medical Research, Brisbane, Australia.The sperm surface protein PH-20 exhibits dual functions, having hyaluronidase activity that enables sperm to penetratethe cumulus and also being involved in zona pellucida binding. The aim of this study was to determine the site of ovinePH-20 expression during spermatogenesis in sheep testes. Fresh testis samples from foetal (n=3), new-born (n=4), 4month-old lambs (n=8), 5 month-old lambs (n=12), and 4-6 year-old rams (n=6) were collected and frozen for totalRNA extraction or fixed in 10% phosphate buffered formaldehyde for histological analysis. A fragment of ovine testisPH-20 cDNA (282 nt) was identified by reverse transcription and polymerase chain reaction (RT PCR) in testis RNA,but not gDNA extracts. The cDNA exhibited 77% identity to human PH-20 and 93% identity to red fox PH-20 eDNAand this sequence was submitted to GenBank (AFI74691). Ovine PH-20 expression was detected using RT-PCR andwestern blotting. The developmental stage of the 4-5 month ram testes was assessed histologically. Ovine PH-20protein was localised in the seminiferous tubules using immunohistochemistry with an anti equine PH-20 antibody.Study of PH-20 rnRNA expression using RT-PCR showed that PH-20 mRNA is only expressed in ram testis and not inthe other sheep tissues including ovary, brain, liver, lung, heart, epididymis, spleen, or kidney. Western blotting (underreducing conditions) revealed a single 74 KDa immunoreactive band in ram testis but not in liver tissue. PH-20 mRNAand protein were identified only in testes where spermatids were present in the seminiferous tubules and in 4-5 montholdram testes undergoing the first wave of spermatogenesis, but not in foetal, new-born or prepubertal testiculartissues. Immunohistochemistry for ovine PH-20 protein in adult ram testis confirmed the cellular localisation in theinitial spermatogenic wave in developing rams. We conclude that ovine PH-20 is expressed post-meiotically in haploidround and elongate spermatids.58. OVARIAN FOLLICULAR DEVELOPMENT IS COMPROMISED IN TRANSGENIC(mREN-2)27 RATS AND ANGIOTENSIN II-INFUSED SPRAGUE DAWLEY RATST.E. de Gooyer*, D.J. KeUyt, R.E. Gilbertt and J.L. Wilkinson-Berka*Departments of*Physiology and tMedicine, St Vincent's Hospital, University ofMelbourne.Angiotensin II (ANG II) is implicated in ovulation, vascularisation and atresia, and may influence these processes by alocal ovarian renin-angiotensin system (RAS), which we have previously localised in rat ovarian stroma and its bloodvessels. The aim of this study was to determine the role ofANG II in follicular development. Three groups of 12 weekold normally cycling, female rats were used: 1) untreated Sprague Dawley (SD), 2) SD infused with ANG II (145ng/kglmin for 12-14 days), and 3) hypertensive homozygous (HMZ) (rnRen-2)27 transgenic rats, which displayamplified extra-renal tissue renin and ANG II. Ovaries were collected at proestrus for histological analysis of folliculartype and number, immunolocalisation of renin and ANG II, in situ hybridisation for ATla receptor (ATla-R) mRNAand radioimmunoassay of ovarian renin content. Systolic blood pressure (SBP; tail cuff) was highest in HMZ Ren-2followed by SD+ANG II and SD. HMZ Ren-2 and SD+ANG II had more antral follicles than SD, but fewer largeantral and pre-ovulatory follicles. Litter size was reduced in HMZ Ren-2 compared to SD. Consistent with the highRAS of the HMZ Ren-2, ovarian renin content and renin and ANG II immunolabelling were increased and ATla-RSD+ANG IT exhibited a reduction in ovarian renin content and ATla-R rnRNA gene expression,rnRNA reduced.indicatin a ne ative feedback on the RAS.N = 4-6 rats er rou SD SD+ANG IISBP (mmHg) 125±1 199±12*Antral Follicles: 150-39011m (% Total) 37.5±4.5 53.5±6.3*Large Antral Follicles: 390-500/lm (%Total) 7.7±1.8 2.4±1.1 *Pre-ovulatory Follicles: >500llID (% Total) 17.1±3.6 3.8±1.5*Litter Size 13.7±O.7 NAOvarian Renin Content (mGU/g) 4.8±0.5 2.7±0.3*ATla-R mRNA (Relative 0 tical Densit ) 75.3±1.8 64.0+2.4*Values are mean ± sem. *p

61. ESTROGEN PROMOTES ANGIOGENESIS IN ERa-EXPRESSING HUMANMYOMETRIAL MICROVASCULAR ENDOTHELIAL CELLS BY UPREGULATINGVEGF RECEPTOR-2Caroline E Gargett, Marina Zaitseva, Kristina Bucak and Peter AW RogersCentre for Women's Health Research, Monash University Department ofObstetrics and Gynaecology, MonashMedical Centre, 246 Clayton Rd, Clayton, Vic. 3168Angiogenesis is the growth of new blood vessels from pre-existing vessels and involves proliferation of microvascularendothelial cells (MEC). VEGF is a major promoter of angiogenesis and mediates angiogenic effects primarily throughinteraction with VEGF receptor-2 (VEGF-R2/KDR) present on the surface of MEC. Estrogen promotes bFGF-inducedangiogenesis, but its effect on VEGF-mediated angiogenesis is unknown. We have demonstrated that MEC derivedfrom human myometrium constitutively express estrogen receptor-~ (ER~), while ERa varies between subjects and isonly expressed in approximately 60% of MEC isolates (1). The aims of the present study were to determine whether (a)estrogen increases VEGF-R2 expression in ERa positive and ERa negative myometrial MEC, (b) ER mediates thiseffect and (c) estrogen promotes VEGF-induced MEC proliferation. Myometrial MEC were isolated fromhysterectomy tissue obtained from ovulating women, purified with DEA-l-coated Dynabeads, cultured and usedbetween passages 1-3 (purity >98% CD31+ cells) (1, 2). ERa and ER~ mRNA were determined by RT-PCR (3) andprotein by flow cytometry or Western blotting. VEGF-R2 expression was measured by flow cytometry using eitherVEGF-R2 antibody or biotin-rhVEGF 165 binding. MEC proliferation was determined by MTS bioassay (2). Estrogen(1 and 10 nM) significantly increased rhVEGF 165 binding and VEGF-R2 receptor expression in ERa positive (P

65. MECHANISMS UNDERLYING THE REDUCED SENSITIVITY TO PROLACTINNEGATIVE FEEDBACK DURING LACTATIONST Anderson!, JL Barclayt, DHL Kusters\ SP Tamt, PLaut, DR Grattan\ MJ Waters 1 ,2, JD Curlewis 11 School ofBiomedical Sciences and 2 Institute ofMolecular Biosciences, University ofQueensland.3 Department ofAnatomy and Structural Biology, University ofOtago, Dunedin, New Zealand.Prolactin negative feedback maintains normal, low prolactin concentrations in non-lactating individuals. Prolactinstimulates the tuberoinfundibular dopaminergic (TIDA) neurons of the hypothalamus to release dopamine whichsuppresses prolactin secretion from lactotrophs. During lactation, however, the sucking stimulus reduces the sensitivityof TIDA neurons to prolactin feedback and consequently high prolactin levels are observed. The mechanisms involvedin this loss of negative feedback are unknown, but may involve changes in prolactin signal transduction pathways.Activation of prolactin-receptors normally results in phosphorylation and nuclear translocation of Stat proteins,particularly StatS, so we investigatedStatS signalling in TIDA neurons of lactating rats. Lactating rats with or withoutpups (removed for 16h) were given a sc injection of either oPRL (250J.lg) or saline vehicle (n=4/treatment), and after Ihwere killed and perfused. Hypothalamic sections were immunostained for tyrosine hydroxylase (Chemicon, MAB318)and StatS (Santa Cruz, Sc835) and confocal images of the dorsomedial arcuate nucleus were captured for quantitation.Stat5 intracellular distribution in TIDA neurons was determined by the ratio of Stat5 fluorescence intensity in thenucleus expressed as a proportion of that in the cytoplasm (N/C ratio). In lactating rats with pups removed for I6h,oPRL injection significantly (P

69. NEGATIVE REGULATION OF PROLACTIN-RECEPTOR SIGNALLING BY SOCSPROTEINS: A NEW MECHANISM IN LUTEOLYSIS?.J.D. Curlewis, M.J. Waters and S.T. AndersonSchool ofBiomedical Sciences, The University OfQueensland, Qld 4072, AustraliaProlactin (PRL) and placental lactogen (PL) play essential roles in maintaining the rodent corpus luteum (CL) throughpregnancy by inducing transcription of some genes (e.g. ERa and ER~, LH-receptor, prolactin-receptor) and repressionof others (e.g. 20a-hydroxysteroid dehydrogenase). These effects are mediated by the PRL-receptor, which signalsthrough several pathways although it is the JAK2/STAT 5 pathway that plays the central role in the CL. At the end ofpregnancy, luteolysis is induced by prostaglandin F2a (PGF2a), which reverses many of the effects of the PRLreceptorsignalling pathway on gene expression. Our recent research on day 19 pregnant rats has shown that one wayin which PGF2a could block PRL-induced gene expression is through inhibition of the JAK2/STAT 5 pathway.Following treatment of rats with cloprostenol, a PGF2a analogue, there is rapid loss of active STAT 5 from ovariannuclear extracts and this effect persists for at least 8 h. Moreover, during this period, treatment with prolactin does notinduce activation of STAT 5, despite the continued presence of PRL-R. The recently discovered Suppressors OfCytokine Signalling (SOCS) have been shown to decrease cell sensitivity to cytokines, including PRL, and so wedetermined whether the negative regulation of PRL-receptor signalling by PGF2a could be due to upregulation ofSOCS proteins. In Northern blots performed on ovaries collected 0.5-16 h after cloprostenol, SOCS-l showed atransient increase at 0.5 h while SOCS-3 mRNA increased 3-fold at 0.5 h and remained elevated until 4 h afterinjection. Western blots confirmed that SOCS-3 protein was increased over the same period and, in ovaries collected 4h after cloprostenol, we confirmed by immunohistochemistry that SOCS-3 was localised predominantly within CLs. Inconclusion, induction of SOCS-3 and to a lesser extent SOCS-l by PGF2a may be an important element in theinitiation ofluteolysis, via rapid suppression of luteotrophic support from PL.70. PARACRINE SIGNALLING WITHIN THE OVARIAN FOLLICLE: POTENTIALFOR TGF-p SUPERFAMILY MEMBERS TO INFLUENCE OOCYTE AND GRANULOSACELL INTERACTIONSRobert B. GilchristReproductive Medicine Unit, Department ofObstetrics and Gynaecology, The University ofAdelaide, TQEH,Woodville, South AustraliaSignalling between the germ cell and somatic cell compartments of the ovarian follicle is a critically important processfor growth and differentiation of the oocyte and the granulosa cells. Communication occurs via paracrine and gapjunctionalsignalling between the two cell types, and indeed both forms of communication are essential for normaloogenesis and folliculogenesis. Traditionally, research has been focused on one side of this communication axis, thatis, on the role of the granulosa cells in nurturing the growth and development of the oocyte. Recent studiesdemonstrate the importance of a bi-directional communication axis, and it is now becoming clear that the oocyte is apivotal regulator of ovarian function. Folliculogenesis fails in the absence of oocyte paracrine signalling, whetherlacking due to genetic deficiencies or from experimental ablation of the oocyte. Oocytes regulate a broad range ofgranulosa cell functions by modulating fundamental control elements; for example, oocytes regulate expression of FSHand LH receptors, kit ligand, inhibin subunits and expression of extra-cellular matrix molecules. As such, oocytes notonly promote growth of granulosa cells and the follicle, but also regulate differentiation processes such assteroidogenesis and physical remodelling of the follicle. The primary recipients of oocyte paracrine signalling are thecumulus cells, which are the specialised granulosa cells immediately surrounding the oocyte. Cumulus cells have adistinct phenotype from the mural granulosa cells (MGC) lining the wall of the follicle, and in fact mairitenance of thecumulus cell phenotype is dependent on oocyte-secretions. In the absence of these oocyte signals the cumulus cellswill spontaneously differentiate toward the MGC phenotype. Oocyte paracrine signalling therefore establishes afunctional morphogenic gradient across the ovarian follicle. The exact identity of these oocyte factors remains elusive,but members of the transforming growth factor-beta superfamily are the prime candidates as some of these growthfactors are able to mimic the effects of oocytes on granulosa cells in vitro. Two family members, which are exclusivelyexpressed in gametes, growth differentiation factor-9 (GDF-9) and bone-morphogenic factor-I5 (BMP-15), are criticalfactors as their absence leads to failed folliculogenesis and complete sterility. Interestingly these molecules have moresubtle effects on fertility with high or low expression of these oocyte factors leading to perturbations such as ovariancysts, anovulation, disrupted oocyte maturation and even increases in ovulation rate and litter size. There is now anemerging appreciation that not only does paracrine signalling from the oocyte allow the oocyte to control its ownmicroenvironment, but oocyte signalling also has profound effects on ovarian function, endocrine well being andfertility.71. EXPRESSION OF TGF-P SUPERFAMILY MEMBERS AND THEIR SIGNALLINGCOMPONENTRY BY THE RAT OVARYAnn E. Drummond, Mitzi Dyson, Minh-Tan Le, Jean-Francois Ethier and Jock K. FindlayPrince Henry's Institute ofMedical Research, PO Box 5152 Clayton, Victoria 3168, AustraliaAn increasing number of transforming growth factor-~ (TGF-~) superfamily members are being shown to exert effectson ovarian function. Most of these factors, which include activin, the bone morphogenetic proteins (BMPs) and TGF-~,act via transmembrane serine and threonine kinase receptors which activate pertinent Smad proteins, complexes ofwhich translocate to the nucleus to mediate gene transcription. Despite understanding the molecular basis of thesesignalling pathways, little has been done to characterise these pathways in the ovary. We've sought to elucidate whichelements of these signalling pathways are present in ovarian follicles of the postnatal rat, as an initial step towardidentifying active pathways in follicle populations. Activin/BMP receptors (ActRIA, ActRIB, ActRIIA and ActRIIB),~glycan and Smad 1-8 mRNAs were expressed by the postnatal rat ovary. Activin receptor and Smad proteins werepresent in oocytes at all stages of follicular development; granulosa cells of primary-antral follicles and theca cells.~glycan protein was present in oocytes, granulosa cells and theca cells at all stages of folliculogenesis. The colocalisationof receptors and Smads supports the notion that activinrrGF~ and BMP signalling pathways may.befunctional in the cellular compartments of the follicle. We hypothesise that follicle populations express elements of theTGF-~ superfamily (and their signalling pathways) in different combinations during development and thus it is theindividual follicle 'blueprint' that determines the response to ligands and ultimately the biological outcomes. Theseconcepts are currently under investigation in our laboratory.Supported by the NH&MRC of Australia (Regkey 983212)72. DIFFERENTIAL GENE EXPRESSION IN INTERLEUKIN-ll RECEPTOR aDEFICIENT MICE COMPARED TO WILD TYPE DURING DECIDUALISATIONChristine White 1 ,2, Lorraine Robb 3 and Lois Salamonsen 1 ,21Prince Henry's Institute ofMedical Research, 2Dept ofObstetrics & Gynaecology, Monash University,3Walter & Eliza Hall Institute ofMedical ResearchDifferentiation of endometrial stromal cells into decidual cells is essential for successful embryo implantation. Femaleinterleukin-l1 receptor a chain (IL-11Ra) null mice are infertile due to disrupted decidualisation, suggesting a criticalrole for IL-l1 and its target genes in implantation. The molecular mechanisms by which IL-11 acts through its receptorto mediate decidualisation are unknown, but it is hypothesised that the expression of genes encoding other regulatorymolecules will be modified by IL-11 signalling. This study aims to identify genes regulated by IL-l1 duringdecidualisation in mouse uterus by DNA microarray analysis, and to examine the expression and functionalsignificance of these genes during early pregnancy. Mice heterozygous for a targeted mutation of the IL-llRa gene(IL-IIRa+/-) were interbred to produce wild type (IL-I1Ra+/+) and null mutant (IL-IIRa-/-) mice. Decidualisation wasinduced in pseudopregnant (plug:::: day 0) IL-llRa+/+ and IL-llRa-/-littermates (n:::: 8/genotype) by oil injection intothe uterine lumen on day 3. Whole uterus was removed at 0, 18, 24 or 48hrs after decidualisation (n :::: 2/group) andtotal RNA prepared by acid-phenol/chloroform extraction. Reference RNA was isolated from wild type unstimulateduterus (n :::: 16). Total RNA (lOllg/sample) was reverse transcribed to fluorescent-labelled cDNA by indirect Cy3 orCy5 coupling. NIA 15K microarrays were probed with experimental (IL-llRa+/+ or IL-llRa-/-) and referenceCy3/Cy5-cDNA and scanned using an Axon GenePix 4000B microarray reader and GenePix Pro 3.0 software. Datawas analysed using R 1.4.1, including normalisation for labelling efficiencies and scanning properties of Cy3 and Cy5,print-tip and spatial effects. At 18 hours after decidualisation, 18 genes were found to be upregulated and 6 genesdownregulated >2-fold in IL-llRa-/- mice compared to wild type. These included expressed sequence tags (ESTs),genes with unknown function in the endometrium, and genes associated with transcription/chromatin, matrix/structuralproteins and angiogenesis. Data analyses for further time points are underway. Genes identified as strongly regulatedby IL-l1 during decidualisation are likely to be important modulators of implantation, and will be further verified andcharacterised. By elucidating the role of IL-11 regulated genes in murine decidualisation, these studies may identifypotential new targets for the manipulation of human fertility.7879

73. MANIPULATION OF BOVINE OOCYTE-CUMULUS CELL GAP JUNCTIONALCOMMUNICATION DURING IN VITRO MATURATION BY MODULATING CELL­SPECIFIC cAMP LEVELSRE Thomas, JG Thompson, DT Armstrong, RB GilchristThe Reproductive Medicine Unit, Department ofObstetrics and Gynaecology, The University ofAdelaide, The QueenElizabeth Hospital, Woodville 5011 SA.Oocyte maturation is associated with the loss of oocyte-cumulus cell gap junctional communication, preventing entry ofmeiotic-modulating factors such as cAMP into the oocyte. Prolonging communication during in vitro maturation(IVM) may improve post-maturation outcomes, such as embryo development, through improved cytoplasmicmaturation. We have previously shown that oocyte and cumulus cell (CC) cAMP levels can be independently regulatedusing selective phosphodiesterase (PDE) inhibitors [1]. The objectives of this study were to examine the effects of celltype-specific PDE inhibitors on the maintenance of oocyte-CC gap junction communication and oocyte meioticprogression. Cumulus-oocyte complexes (COC) were aspirated from antral follicles of abattoir-derived ovaries.Oocyte-CC gap junction communication during oocyte maturation was measured using the fluorescent dye, calcein­AM. COCs were cultured in the presence of specific PDE inhibitors; milrinone (MR, oocyte PDE3 inhibitor) orrolipram (RP, CC PDE4 inhibitor), and were pulsed with calcein-AM to allow dye transfer between the two cell types.Following CC removal, fluorescence in denuded oocytes was measured by microphotometry and meiotic progressionassessed. In control COCs, dye transfer from CC to the oocyte fell progressively from 0 to 8h, after which oocyte-CCgap junction communication was completely lost. From 3-6h of maturation, loss of gap junction communication wassignificantly (P

77. INSEMINATION AND UTERINE LYMPHOCYTE TRAFFICKING IN MICEJohn J. Bromfield and Sarah A. RobertsonDepartment ofObstetrics and Gynaecology, University ofAdelaide, SA 5005Lymphocyte populations facilitating embryo implantation and early placental development have immunosuppressivephenotypes, similar to those mediating immune tolerance in other mucosal tissues. It is unclear how and when thesepopulations become activated, but they have some similarities to those primed earlier in pregnancy at insemination.The mechanisms defining lymphocyte phenotype and homing patterns occur at activation, and lymphocytes primed inspecific tissues have a propensity to recruit back to those tissues [1]. Moreover, paternal and other antigens are sharedby semen and the conceptus. Thus we hypothesise that mating comprises the 'priming' event for activation oflymphocytes required for embryo implantation [2]. The aim of this study was to investigate whether insemination leadsto the activation of lymphocytes that can be recruited preferentially into the implantation sites of pregnant mice. Weemployed cell tracking techniques, utilising [125l]-iodo deoxyuridine C 25 IdUR), to label lymphocytes recovered fromthe para-aortic lymph nodes (PALN) which drain the uterus, in virgin mice or 4 days after insemination. Labelled cellswere transferred to day 6 pregnant recipients via the lateral tail vein, and recruitment into a range of tissues wasassessed by y counting 24 hours after transfer. Tissue specific patterns of lymphocyte recruitment were identified, withpreferential recruitment of PALN lymphocytes into lymphoid and uterine tissues. PALN lymphocytes from mateddonors showed a pattern of recruitment into the uterus, preferentially into implantation tissues as opposed to interimplantationtissues (p=O.OI), which was not explained simply by the increased vascularity of this site. The distinctionwas not evident when PALN lymphocytes were recovered from virgin donors. PALN lymphocytes from virgin donorsalso showed a greater propensity to recruit back to the PALN and other lymphoid tissues (p

81. INTRAOVARIAN REGULATION OF FOLLICULOGENESIS: SIGNAL MOLECULESAND THEIR CELLULAR SOURCESDavid T ArmstrongReproductive Medicine Unit, Dept ofObstetrics and Gynaecology, University ofAdelaide, SAFolliculogenesis in mammals is a continuous and dynamic process that begins during embryonic development in uteroand continues until the end of adult reproductive life. The overall process is regulated by a complex array of interactingchemical signals originating both within and external to the ovaries. The identities and relative importance of specificsignals change as follicle growth progresses through a series of morphologically distinct stages, beginning withinitiation of growth of the dormant primordial follicles, and ending in either atresia or ovulation. The initiation offollicle growth occurs when a primordial follicle is activated by as yet unknown mechanisms and enters a pool ofgrowing follicles. Recent evidence has established the essential role of the oocyte as the driving force in promotingfollicle growth throughout pre-antral and early antral stages, mediated via recently identified members of the TGF~superfamily of growth factors, GDF9 and GDF9B, which have essential roles in both proliferation and differentiationof granulosa cells. The follicular somatic cells, including both granulosa and thecal cells, also contribute to regulationof follicle growth during these stages, through secretions of several other peptide growth factors, in particular bFGF,IGF-I, TGF~, activin, and inhibin. Upon reaching antral stage of development, follicles acquire responsiveness tofollicle-stimulating hormone (FSH) and luteinizing hormone (LH), and these gonadotrophins then assume primaryimportance in regulation of growth to the pre-ovulatory stage, with growth factors continuing to play supportive andmodulating roles. The actions of FSH and LH are mediated in part via androgens and oestrogens secreted by the thecaland granulosa cells, respectively. With the approach of ovulation, migratory cells derived from the circulation,including macrophages and to a lesser extent other leukocyte subsets, infiltrate the ovarian stroma and thecal layer ofthe pre-ovulatory follicle, where they play essential roles in the tissue remodelling that leads to rupture of the follicleand its transformation to a corpus luteum at ovulation. Their actions are mediated via pro-inflammatory moleculesincluding cytokines, chemokines and eicosanoids secreted by the macrophages and by the follicle cells themselves.Recent evidence implicates signals initiated in the endometrium in response to semen exposure, in enhancing thisleukocytic infiltration of the ovary, and influencing their pattern of cytokine secretion, with important consequences inenhancing ovulation and luteinization.82. OESTROGEN TARGET SITES IN HUMAN TESTISPTK Saunders, TL Gaskell, JE Sierens, MR Millar, RM Sharpe and GA ScobieMRC Human Reproductive Sciences Unit, Edinburgh University Medical School, Edinburgh EH16 4SB, UKIt has been proposed that oestrogens play a role in regulating germ cell function in adulthood and during fetal life.Oestrogen action is mediated via high affinity intracellular receptors expressed in target tissues [1]. Two subtypes ofoestrogen receptor known as ERa (NR3A1) and ERp (NR3A2), have been cloned and hER~ variant isoformsidentified. In target cells these receptors can exist as homo- or heterodimers [1]. We have used immunohistochemistryto examine the patterns of expression of ERs in human fetal and adult testis to determine the cellular targets foroestrogen action. In parallel studies we have prepared ER constructs and used these to examine steroid binding to ERhomo- and heterodimers in transfected cells.In human testicular tissues (fetal or adult) we have never detected ERa mRNA or protein. With a polyclonal antibodywe detected ER~ protein in multiple testicular cell nuclei including those of Sertoli cells, Leydig cells and peritubularmyoid cells as well as some germ cells [2]. Additional studies using monoclonal antibodies capable of discriminatingbetween human wild type ER~ and the human ER~cx/~2 [3] splice variant revealed discrete patterns of expression ofthe subtypes. In adult testis immunoexpression of wtER~ was most intense in pachytene spermatocytes and roundspermatids, whilst low levels of expression were detected in Sertoli cells, spermatogonia, preleptotene, leptotene,zygotene and diplotene spermatocytes. Expression of ER~cx/P2 protein was highest in Sertoli cells and spermatogoniawith low/variable expression in preleptotene, pachytene and diplotene spermatocytes [4]. In the fetal testis wtER~ wasundetectable in gonocytes whereas these cells expressed the highest levels of ER~cx/~2 compared with other testicularcell types. Both ERp1 and ER~cxl~2 were detected in some but not all Sertoli cells, peritubular cells and Leydig cells.Transfection studies with ERa and ER~ constructs confmned that wtER~ and ERa were activated following exposureto oestogenic ligands but that the ER~cx/~2 variant did not bind oestrogens and was not capable of activating an EREcontainingreporter construct.The testicular cells most likely to be targets for oestrogens in adulthood are round spermatids in which levels ofexpression of wtERp1 are high. In contrast, expression of ERcxlP2, an isoform that may act as a dominant negativeinhibitor of ER action, in adult Sertoli cells and spermatogonia, and in fetal gonocytes could prevent these cellsresponding to oestrogens.1. Nilsson, S., et al., Phys Rev, 2001. 81: 1535. 2. Saunders, P.T.K., et al., Mol Hum Reprod, 2001. 7: 227. 3. Ogawa,S., et al., NAR, 1998.26: 3505.4. Saunders, P.T.K., et al., JCEM, 2002. 87: 2706.8483. MALE CONTRACEPTION - APPROACH TO HORMONAL METHODSRobert I McLachlanPrince Henry's Institute ofMedical Research, Melbourne VictoriaWorldwide, men particip~te actively in contraception through natural methods, condo~ usage and vasectomy but no new,acceptable, safe and reverSIble methods have become ~v~lable. Male hormonal contraception (MHC) is the most promising avenueof research. T~stosterone (T). treatment suppresses pItUItary LH and FSH secretion, the primary endocrine signals required forspermatoge~esI.s. Large mUltIc~ntre WH~-sponsored trials have shown that high doses of T reduce sperm counts to zero(azoospeI1~l1a) In -70% CaucaSIan men whIle over 90% have sperm counts < 1 million/m (severe oligospermia), a level at whichcontraceptIve efficacy appears adequate. ~ut the need for frequent injections and androgenic side effects (acne, mood changes,redu~ed ~DL-cholesterol) demand al~ernat~ve methods of T administration. Testosterone implants provide the prototype for morephySIOlogICal T treatm~nt .but pellet InsertI.o~extrusion limit their wide application. New long acting intramuscular preparations,such as T undecanoate III 011, are most promlSlng and may allow 8-12 weekly injection intervals.Co-admi~istration of .other gonadotropin-suppressing agents, such as GnRH antagonists or progestins, enhance spermatogenicsu~pressI~n .and permI.t the use of lower T doses. In MHC, progestin action is probably via gonadotropin suppression but directactIons WIthIn the testIS have not been excluded. A range of progestins have been used including levonorgestrel desogestrel anddepot med:oxy-prog~sterone acetate, and studies have shown that -75% and -95% of men become azoospe~c or severelyohgospermIc, respectively. The only T plus progestin efficacy study to date has reported excellent contraceptive efficacy over a 1year ~xposure (Tu~er et al, ESA 2002, a~stract). The stage is set for pharmaceutical company involvement, sadly lacking in thepast, In order t~ b~ng the first MHC regImen to market. About 5% of men fail to suppress adequately and understanding thereason(s) for theIr faIlure to respond is important in gaining the widest acceptance of MHC.We have explored the MHC effects on sp~rmatogenesis in men using stereological techniques for germ cell quantification andshowed that. type Apale--+B spermatogomal development and sperm release (spermiation) are the key sites of action. Thesperm~to~omal. defe~t probably results from FSH withdrawal but the subtypes affected and their mechanism of action are unknown.SpermIatIon faIlure IS a feature of both acute (accounting for dramatic falls in sperm output within 3 weeks) and chronic MHCtreatment. Both FSH and T regulate rodent spermiation but nothing is known in man. Cell-cell adhesion/communication andregulation of gene expression in later germ cell types can now be explored using laser capture microdissection and in vitro culture.F~om an endocrine regulatory viewpoint, MHC suppresses serum LH to 0.05). In ~ll treatment groups pnm?rdIal follicles were the most abundant follicle type, however primary andsecondary f~lhcles were also observed m grafts from all treatment groups. This study confirms that xenograftingsup?orts folhcular development within wallaby pouch young ovarian tissue (1-2). Interestingly, follicle survival andfolhcular development was sustained in all treatment groups, which indicates that the hormonal environment of therecipient may not playas crucial a role as previously believed. [1] Mattiske et al (2001) Proc Aust Mamm Soc 47:35.[2]Mattiske D et al (2002) Reproduction 123:143-153. [3]Cox S-L et al (2000) Fertil Steril74:366-371.85

85. DEVELOPMENT OF ANTRAL FOLLICLES IN RABBIT OVARIAN XENOGRAFTSIN IMMUNODEFICIENT MICEK. Muirhead l , S-L. Cox 2 and M. Holland l .1Pest Animal Control Cooperative Research Center, CSIRO Sustainable Ecosystems, GPO Box 284, Canberra, ACT2615, 2Department ofPhysiology, Monash University, Clayton 3800The primordial follicle pool is established in mammalian ovaries in v~ry early ~ife. Fro~ ~is. time forward thepopulation of primordial follicles progressively decreases as a result of eIther atre~Ia or th~ ImtlatlOn of g~owt~ anddevelopment. Very little is understood about the processes that regulate normal follIcle recrU1tme~t and .survIVal ~n therabbit. This report describes an initial trial in the use of ovarian xenografting as a means to mvestlgate folhculardynamics in juvenile rabbits. Fresh ovarian tissue from NZ White rabbits aged 2, 4, 8 and 12 weeks old were graftedunder the renal capsule of ovariectomised NOD-SCID mice (n=35). Grafts were recovered 8 days, 2 weeks, 4 weeks or8 weeks later, assessed macroscopically and subjected to histological evaluation. All grafts were present and hadbecome vascularised at the time of autopsy. Grafts from two-week-old donor rabbits recovered 2 and 4 weeks aftergrafting did not contain any follicles. In contrast four-week-old donor rabbit tissue (grafted for.4 weeks) containe?healthy looking primordial and preantral follicles, resembling folliculogenesis normally observed m 8-week-old rabbItovaries. Similarly four-week-old donor tissue (grafted for 8 weeks) contained follicles at all stages of developmentincluding antral follicles, resembling 12-week-old normal rabbit ovaries. Whilst tissue from 8 week old donors (graftedfor 4 weeks) displayed apparently normal primordial, preantral and antral follicles (diameters of approximatel~ 1rnn:),tissue grafted for only 8 days and 16 days contained primarily degenerating preantral follicles and healthy pnmordlalfollicles. Interestingly, corpora lutea like structures were detected in a number of grafts from donors aged 12 w~eksthough "ovulation" of normal oocytes remains unconfirmed. In general the dev~lopment a~d morpholo~y of folhcleswithin the xenografted rabbit ovarian tissue resembled the normal pattern of folhculogenesls. Thus ovarian xenograftsin NOD-SCID mice can serve as an experimental model for juvenile rabbit follicular growth and development. Weintend to use this technique in conjunction with an in vitro culture system we have developed for rabbit ovarian corticalexplants to investigate factors regulating folliculogenesis.86. ACUTE FSH WITHDRAWAL IMPAIRS SERTOLI AND GERM CELLDEVELOPMENT IN THE IMMATURE RATSarah J. Meachem, Jessica Ziolowski# and Kate L. Loveland#Prince Henry's Institute ofMedical Research, Clayton, Victoria, 3168#Monash Institute ofReproduction and Development, Monash University, Clayton, Victoria, 3168Pituitary follicle stimulating hormone (FSH) is required for normal testicular developmen~, affecting both Serto.li ~dgerm cells (l) through its receptors on Sertoli cells, and thereby determining the po~ent1al for sperm productlO.n madulthood. The impact of FSH on in vitro Sertoli cell proliferation alters during maturatIOn (2), though the mech~ms~sunderpinning these actions and their development are not understood. To establish an in ~ivo model for examm~tlOnof the role of FSH at discrete developmental stages, we assessed the impact of acute FSH wIthdrawal on the Sertoli andgerm cell populations during early postnatal life. Sprague Dawley rat pups (n = 7/grou~) ~e.re ~assively immunisedagainst FSH using an ovine polyclonal antibody to rat FSH (10 mg/kg by subcutaneous daIly mJect~on) for 2 and ~ daysprior to death on days 3, 9 and 18 days postpartum. Control animals recei~ed a normal. sh~ep lmmunoglobulm ~10mg/kg). Testes were immersion fixed with Bouin's fluid and embedded in resm for det~rnun~tlOn of cell num~er us~ngthe optical disector stereological technique, and paraffin for assessment of prolif~ratlOn and apopto~lS usmgimmunohistochemistry. Sertoli and germ cell numbers per testis were unchanged followmg 2 days of FSH WIthdrawalat day 3, 9, and 18 compared to controls. In response to 4 days of FSH withdrawal in day 9 rats, Sertoli cell an~ typeA/intermediate spermatogonial number were decreased to 73% (P

89. IMMUNOCONTRACEPTIVE EFFECT OF RECOMBINANT PORCINE ZONAPELLUCIDA C PROTEIN IN RABBITSE McLaughlin\ AJ Robinso~,B Inglis 2, J Swan 2 , M.K. Holland 2 & PJ Kerr 21School ofEnvironmental & Life Sciences, University ofNewcastle, Callaghan NSW 2308& 2Pest Animal Control CRC, CSIRO Sustainable Ecosystems, GPO Box 284, Canberra, ACT2615.Despite the success of biocontrol methods of population control (such as calicivirus) rabbit populations within Australiaremain a significant problem particularly in temperate zones. An alternative is to induce sterility in the female rabbitpopulation via immunisation with a viral vector containing an ovarian antigen. Fertility trials using total porcine zonaepellucida as an immunogen have been shown to induce infertility in a number of diverse species. Here we describe theproduction of recombinant porcine Zona Pellucida C (pZPC) protein and its use as an immunocontraceptiveimmunogen in small scale fertility trials in domestic rabbits. Recombinant protein was produced by expression of thefull length porcine ZPC cDNA in a vaccinia virustr7 system in a co-infected monkey kidney cell line (CVl) andpurified using lectin affinity chromatography. Two adult male and ten adult female laboratory rabbits were inoculatedwith IOOllg recombinant pZPC in Freunds Complete adjuvant and boosted twice with 50llg in Incomplete Freundsadjuvant on Days 28 and 56. Sera were collected prior to immunization and following boosting (Days 14,44, 70). Fourmale rabbits of proven fertility were used to mate both immunized and untreated control rabbits (n=4). The immunizedrabbits were sacrificed 9 days post mating and the reproductive tracts examined for corpora lutea and implantation sacs.Ovaries from rabbits immunized with recombinant pZPC were fixed in Bouin's solution. Tissues were paraffinembedded and sectioned prior to staining with hematoxylin and eosin. By Day 70 sera from all animals containedantibody capable of detecting recombinant pZPC protein by Western blotting. Immunoperoxidase staining was detectedon the zona pellucida of early primary and more mature follicles when the antisera from immunized animals were usedto probe normal ovarian sections. At autopsy all ten immunized animals had visible corpora lutea but only two werepregnant with 10 and 2 viable implantations respectively. The remaining eight females displayed signs ofpseudopregnancy only. In contrast all control females (n=4) had normal sized litters (mean±sem. 7.25±1.3).Histological evaluation showed no difference in ovarian folliculogenesis between treated animals and age matchedcontrol specimens. Serum antibody titres measured using ELISA showed no difference between males and femalesimmunized with pZPC and no correlation with pregnancy. As antibody was not detected bound to the ZP in vivo themechanism by which recombinant pZPC induced infertility remains to be elucidated.90. EFFECT OF ANGIOGENESIS INHIBITORS ON OESTROGEN-MEDIATEDENDOMETRIAL ENDOTHELIAL CELL PROLIFERATION IN THEOVARIECTOMISED MOUSEB Heryanto\KE Lipson 2 , and PAW Rogers!lCentrefor Women's Health Research, Monash University Department ofObstetrics & Gynaecology, Monash MedicalCentre, 246 Clayton Rd, Clayton, Victoria 3168, Australia2SUGEN, Inc., 230 E. GrandAve, South San Francisco, CA,94080, USAIt has been suggested that endometrial angiogenesis in response to the sex steroids oestrogen (E) and progesterone (P)is mediated at a local level via compounds such as vascular endothelial growth factor (VEGF), fibroblast growth factor(FGF), and platelet-derived growth factor (PDGF), acting through their respective tyrosine kinase receptors (RTK). Theaim of the present study was to use RTK angiogenic inhibitor compounds to determine if VEGF, FGF or PDGF playarole in mediating endometrial endothelial cell (EC) proliferation following administration of E and P. Endometrial ECproliferation was induced in adult ovariectomized mice by either E alone for 1 day (EI), or P with low dose E followedby P with high dose E (PE). Each angiogenesis inhibitor compound was injected daily for four days (lOOmg/Kg/day,s.c.) before endometrial tissue collection. We also evaluated the effect of VEGF antiserum (0.2ml i.p.) on ECproliferation at E1. All the angiogenic inhibitor compounds significantly reduced EC proliferation activity at El andPE. The greatest reduction in EC proliferative index was at EI in the group treated with a VEGF receptor inhibitor (2.5± 0.7% versus 27.9 ± 1.1 %, P

93. MAST CELL NUMBER AND SPATIAL LOCATION IN THE VAGINAL CUL-DE-SACOF THE BRUSHTAIL POSSUM AFTER ADMINISTRATION OF EXOGENOUSOESTRADIOLPM Mahonel, PR Hurse, BJ McLeod!,2, MA McConnele1Department ofAnatomy and Structural Biology; 3Department ofMicrobiology, University ofOtago, Dunedin, NewZealand. 2AgResearch Invermay Agriculture Centre, Mosgiel, New ZealandMast cells are present in large numbers in the vaginal cul-de-sac, a component of the reproductive tract in femaleBrushtail possums. Their presence does not appear to be an immunological response to the presence of microflora, asestablished in a previous study of mast cell number and spatial location in cul-de-sac tissues (1). In this study mast cellnumbers and spatial location were determined in seasonally anoestrus possums and those treated with sesame oil oroestradiol 17~ implants for 6 days.Cul-de-sac tissue was collected aseptically for microbiological analysis, and the Fractionator and Optical Disectorstereological methods were used to quantify mast cell populations in tissue sections.In all groups (n=6/group), microflora populations were consistently low «4.0 x 10 4 organisms gram-I). Mean weightand volumes of epithelial and connective tissues were significantly greater in oestradiol treated animals). Meannumbers of mast cells in epithelial and connective tissues did not differ significantly amongst groups. Mean mast celldensity was therefore significantly greater in the seasonally anoestrus and control animals than the oestradiol treated.Although previous emperical studies may have reported a decrease in mast cell number when circulating oestrogenlevels were elevated (2, 3) this study has highlighted the need for careful interpretation of both tissue volume and cellnumber (4).References(l)Mahoney et al., (2002) (in press). (2)Krishna et al., (1986) Journal of Reproduction and Fertility 32:1211-7.(3)Wilhelm et al., (2000) Endocrinology 141:1178-86. (4)Mayhew TM and Gundersen HIG (1996) Journal ofAnatomy 188:1-15.94. IMMUNOLOCALISATION OF INTERLEUKIN 11 AND ITS RECEPTOR INCYCLING ENDOMETRIUM AND IMPLANTATION SITES OF THE RHESUS MONKEYE. Dimitriadist, L. Robb 2 , v-x Liu 3, W. M. Carter 2 , H. M. Martin 2 , L. A. Salamonsen!Prince Henry's Institute ofMedical Research, Clayton, 3168 1 , The Walter And Eliza Hall Institute ofMedical Research,PO Royal Melbourne Hospital, Melbourni, Institute ofZoology, Beijing, 100080, China 3 .Interleukin 11 (IL-l1) signalling is essential for stromal cell decidualization and embryo implantation in the mouse 1and is involved in decidualization in the human 2 . IL-ll action is mediated via binding to its specific IL-l1 receptoralpha (IL-I1Ra). The aim was to examine the temporal and spatial location of IL-l1 and IL-I1Ra in cyclingendometrium and implantation sites of the Rhesus monkey. Immunohistochemistry was performed on cycling monkeytissue from proliferative phase day 13 (one day before ovulation) and secretory phase days 19,24 and 29 (5, 10, and 15days respectively after ovulation) (n=4 per day), and from implantation sites between days 24 and 35 of pregnancy(n=8). Data was expressed as relative intensity units. IL-ll immunoreactivity was elevated in the secretory phase ofthe menstrual cycle compared to the proliferative phase. At day 24 of the cycle, both IL-ll (2.8±1.2, mean±SEM) andIL-llRa (3.4±0.8) staining were most intense in glandular epithelium. Staining in the stroma for IL-ll was minimal(0.4±0.1) and moderate for IL-llRa (1.3±0.3). By day 29 of the cycle staining for IL-ll was higher in stromacompared to glandular epithelium (3.4±0.8 vs 1.7±0.6, P

-AUTHOR INDEXAuthorAbstract No.Aitken, RJ 5,9,54,56 Duckworth, J 28Aloe, J 78 Dyson, M 35, 71Amman,G 63Anderson, ST 65,69 Ecroyd, H 54,88Armstrong, DT 40,48,73,81 Eriksson, BM 19Ashman, RJ 14 Ethier, J-F 71Aspden, WJ 24,37 Evans, G 19,21,22Asquith, K 54Assinder, S 49,53 Fenwick, MA 59Findlay, IK 35,62,71,92Baker, MA 9 Fink, J 53Barclay, JL 65 Fisher, PI 17Bathgate, R 19 Fleming, JS 27,57Beagley, KW 88 Forbes, E 47Blewitt, M 2Breier, B 15 Galloway, SM 80Britt,K 62,63 Gargett, CE 61Brockway, A 20 Garry, PG 42Bromfield, JJ 77 Gaskell, TL 82Bucak, K 61 Gibbs, GM 8Bunn, SJ 42 Gilbert, RE 58Byers, S 38 Gilchrist, RB 39,40,70,73Grattan, DR 65Cakouros, D 68 Green, DPL 36Carter, WM 94 Gregan, S 80Cetica, P 39 Gromoll, E 6Chong, S 2 Grupen, CG 14Chrousos, GP 66Clarke, HG 38 Hallett, A 29Clarke, II 43 Hamilton, HM 16Clyne, CD 64 Handelsman, DJ 47Collett, RA 13 Hanrahan, SP 80Conway, AJ 47 Harding, R 95Cook, S 78 Hartwich, KM 16Cotton, LM 8 Harvey, AJ 12,13Cowan, P 28 Hedger, M 45,51Cox, S-L 84,85 Herlihy, A 55Crone, J 63 Herrid, M 7Cui, X 28 Heryanto, B 90Curlewis, JD 65,69 Hessian, PA 36Hickox, D 55Daish, T 68 Hill, JR 17Davies, CJ 17 Holland, MK 1,85,89Davis, GH 80 Hollinshead, FK 21,22de Gooyer, TE 58 Hope, SA 38de Kretser, DM 8,44,45,51,55,87 Hospers, M 95Dekker, GA 75 Hurst, PR 27,42,59,93Dent, L 29Dharmarajan, AM 67 Inglis, B 89Dimitriadis, E 94 Ingman, WV 50D'Occhio, MJ 24,37 Irving-Rodgers, HF 37,60Dorstyn, L 68Druker, R 2 Jasper, MJ 26Drummond, AE 35,44,62,71 Jenkin, G 8492 98

Jones, D 28 Montgomery, GW 57 Sawyer, H 80 Thomas, RE 73Jones, M 62 Morrison, JR 8 Schlafer, DH 17 Thompson, JG 12,13,39,73Jones, RC 56,88 Morrissey, MP 40 Schlatt, S 32 Tilbrook, AJ 43Juengel, JL 80 Moss, TJM 95 Scobie, GA 82 Torres, CAA 18Muir, J 45,51 Scobie, S 28 Tremellen, KP 33, 75Kakar, MA 23 Muirhead, K 85 Scott, CJ 43 Trigg, TE 24Kaye, PL 11 Murata, Y 63 Sebire, K 8,55 Trounson, A 30Keelan, J 15 Sferruzzi-Perri, A 29 Tumer,L 47Kelly,DJ 58 Newcombe, N 88 Sharkey, DJ 75Kelly, JM 16 Newnham, JP 95 Sharpe, RM 82 Waddell, BJ 74Kerr, PJ 89 Nicholson, HD 49,53,57 Shaw, G 25,96 Wade,MA 56Kind, KL 12, 13 Nie,G 92 Shaw, J 20,84 Waldhauser, F 63Kleemann, DO 16,23 Nieschlag, E 6 Short, RV 87 Walker, SK 16,23Knee, RA 88 Nitsos, I 95 Sidhu, KS 10 Waters, MJ 65,69Kovacic, A 64 Nogoy,N 52 Sierens, IE 82 Wells, D 15Kovacs, J 63 Nogueira, ET 18 Simoni, M 6 Weston, GC 91Krupa,M 60 Norman, RJ 26 Simpson, ER 62,63,64 White, C 72Krutskikh, A 9 NottIe, MB 14 Smith, JF 52,57 Whitelaw, E 2Kumar, S 68 Smith, JT 74 Wijayanti, GE 25Kusters, DHL 65 O'Brien, JK 21,22 Snow,M 84 Wilkinson-Berka, JL 58O'Bryan, MK 8,55 Speed, CJ 64 Williams, PJ 4Lau,P 65 Ochsenktihn, R 51 Stanton, JL 36 Wilson, T 80Le, M-T 71 O'Connor, A 45 Stephenson, T 46 Wishart, S 47Lee,R 15,52 Okuma, Y 45 Sun,M 49Lepore, D 78 O'Leary, S 48 Sutton, M 39 Yu,F 57Li,A 76 O'Neill, C 76 Swan, J 89Lipson, E 90 Zaitseva, M 61Liu, PY 47 Parry,L 53 Tam, SP 65 Zhang, Z 87Liu, Y-X 94 Paya, K 63 Tan, OL 27 Zhou, J 64Lorimer, MF 23 Peterson, J 15 Temple-Smith, PD 41 Ziolowski, J 86Loveland, BE 31 Peura, TT 16 Thomas, P 78Loveland, KL 44,86,87 Phillips, D 45Luetjens, CM 6 Preis, J 2Luu,K 92Quinn, MCJMacmillan, K 2036Maddocks, S 23 Rakyan, V 2Mahoney, PM 93 Rath,D 22Mancio, AB 18 Ravelich, S 15Markham, K 11 Renfree, MB 25,41,87,96Martin, GB 18 Richings, NM 41Martin, HM 94 Ritter, LJ 26,40Martin, L 3 Robb,L 72,94Mate, KE 10 Roberts, CT 95Mattiske, D 84 Robertson, DM 46Maxwell, WMC 19,21,22 Robertson, SA 29,34,48,50,75,77McConnell, MA 93 Robinson, AJ 89McDonald, R 57 Rodger, JC 10McFarlane,. J 7 Rodgers, RJ 37,38,60McIlfatrick, SM 14 Rogers, PAW 61, 90, 91McLachlan, RI 46,47,83 Roman, SD 56McLaughlin, E 89 Ross, S 78McLeod, BJ 53,93 Rudiger, SR 16,23McNair, L 96 Ryan, NK 26McNatty, KP 80Meachem, SJ 86 Salamonsen, LA 72,92,94Meehan, T 44, 87 Santiago, LL 18Meyers, SA 57 Saunders, PTK 82Millar, MR 82 Sawyer, DE 594 95

start time8:30am9:00am9:30am10:00am10:30am11:00am11:30amnoon12:30pm1:00pm1:30pm2:00pm2:30pmSunday22ndESAESAlSRB PROGRAM AT A GLANCEMonday23rdESAJapan/Australiaplenary,Hall 0, start8:45amServierAwardInvsetigatorScientistAwardHall 0SRBFoundersLectureHall 0SRBOrals #1&2Rooms 2 &10, start8:45amOrals #3&4Rooms 2 &10Lunch & Poster viewing* Posterdiscussion#1,2,3,4&5until 3:45includesafternoonteamorning teaSRBSymposiumRoom 2Symposium,Hall 0* FreeCommun.Rooms 11 &2HarrisonLectureHall 0Lunch & Poster viewing* Posterdiscussion#6,7,8,9,10 & 11until 3:45includesafternoonteamorning teaSRBSymposiumHall 0Wednesday25thESAJointADS/ESAplenary, HallB/C*SRBSymposiumRoom 2* ESAIADSSymposiumHall B/C* ESASymposiumRoom 1morning teaafternoon teaSRBSRBSymposium,Hall 0Lunch & Poster viewingposter Orals # 9 &viewing 10--..,...--,-__--l Rooms 1 & 2ESAIADSPlenaryHall B/Cstart time3:00pm3:30pm4:00pm4:30pm5:00pm5:30pm6:00pm6:30pm7:00pm7:30pm8:00pmSunday22ndESASRBSymposiumRoom 2Welcome FunctionExhibition FoyerWomen in EndocrinologyRiverview 3 (starts 7pm)Monday23rdESA* FreeCommun.(rooms 4 &10) and Jointreproductivesession (HallD)ESAAGM,Hall 0, starts5:45pm,SRBSRBafternoonteaOral #5Room 2andJointReproductive sessionHall 0SRB StudentMeeting,AdelaideClubStudent BBQ, AdelaideUniversity ClubGold Coast RoomTuesday24thESA* FreeCommun.(rooms 11 &10 and HallD)SRBSRBafternoonteaSRB JuniorScientistAwardFinalistsRoom 1Taft Lecture, SRB AGM,Hall 0 Room 1Conference DinnerStamford Grand Gelenelg,catch tram at 7pmWednesday25thESAESASymposium,Room 1ESAIADSsymposium,Hall B/CSRBESAlSRBlate breakingscience,Room 296 97