ABDBBCABAPutative model of how cross-linked WDR54 inhibits the internalization of the activated EGF receptor. TG2 (transglutaminase 2), Ub (ubiquitin), pY (tyrosine phosphorylation)BWhatfactorsarerelatedinfolliclegrowth?34Keywords: Oogenesis, Folliculogenesis,In vitro germ cell culture, Germ cell preservation,Animal reproductionResearch keywords?Signal transduction?Protein phosphorylation?Knock out mouse?adaptor proteinTheobjectiveofourlaboratorymainlyfocusesonunderstandingoogenesisandfolliculogenesisinmammalianovarybygeneratinginvitroculturemethodfortheirliveimagingsystem.Backgrounds:Inmammalstheovarycontainsmorethan1millionoocytesatthemaximum.However,onlylessthan1%ofoocytesareovulatedduringreproductiveperiod,whereasmostofoocytesdegenerateinthecourseoffollicledevelopment.Thequestionsare;Howaretheovulatedoocytesselectedduringfollicledevelopment?Whatfactorsarerelatedinfolliclegrowth?Ourgoalsaretomakeclearthesequestionsandapplyournewfindingstolivestockreproductionandhumaninfertility.WehaveidentifiedmanyfunctionalmoleculesactingdownstreamoftounderstandthetheEGF(epidermalgrowthfactor)complicatednetworkofsignalingbyphospho-proteomics.Wewillespeciallyfocusonthefunctionofthenewlyidentifiedproteinsthroughthisanalysis.ThecurrentdeterminednumberofunknownmoleculesintheEGFreceptor-mediatedsignalingpathwayisover15.Ourgoalinthenearfutureistoestablishthefunctionofeachmoleculeinthispathwaybybiochemical,molecularbiologicalandcellbiologicalmethods.Research themes1.Toidentifythemolecularmechanismsofoocytedevelopmentandselectioninmammalianovaryusinglivecellimaging.2.Todevelopanewculturemethodandchemicaldefinedmediumforinvitroovaryandfolliclecultures.3.Toidentifythemolecularmechanismsofembryoimplantationandplacentaformation.GAREM projectTheGAREMfamilyofproteinsthatincludetheubiquitouslyexpressedGAREM1andbrain-specifictype,GAREM2,havebeenrecentlycategorizedasnoveladaptorproteins.CLPABP projectA PH domain-containing protein, CLPABP, plays a role in mRNA metabolism, such as its stability or instability, translation regulation, and transport in the cytosol.WDR54 projectWDR54 is a member of theWD40 repeat domain-containing protein family. This protein is cross-linked to tightly form both dimers and trimers, and that it has a regulatory effect on Erk.receptoritsintracellularA:SchematicrepresentationoftheofCLPABP.Thenumberofaminoacidtheonboundariesisindicated.primarystructureresiduesB:CLPABPKOmicewerenormalatbirthandgrewwhennormallyyoung.However,oldmalemicepresentedafattyphenotype,similarinmetabolicsyndrome,seentothatHistologicalsectionsofmouseovarycollectedat10dayafterbirth.A:Eachoocyteisenclosedinsinglefollicle,andoocytesaregrowingconcomitantlywithfolliclegrowth.B:EnlargedfigureofA.Alotofprimordialfolliclesatthesmallestfolliclestageexistasaprimordialfolliclepool(dashedcircle).50?mKanako MOROHAKUAssistant Professor, Ph.D.Hiroaki KONISHIProfessor,Ph.D.How are the ovulated oocytes selected?WearegoingtoidentifythemechanismsofCoogenesisandfolliculogenesisusinginvitroovaryculture.AMost of oocytes degenerate in the course of development.Laboratory of Germ Cell Physiology and EngineeringBiotechnology DivisionBiotechnology DivisionLaboratory of Biochemistry
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