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ABDBBCABBWhatfactorsarerelatedinfolliclegrowth?APutative model of how cross-linked WDR54 inhibits the internalization of the activated EGF receptor. TG2 (transglutaminase 2), Ub (ubiquitin), pY (tyrosine phosphorylation)31Laboratory of Germ Cell Physiology and EngineeringBiotechnology DivisionHiroaki KONISHIProfessor,Ph.D.Biotechnology DivisionLaboratory of BiochemistryKanako MOROHAKUAssistant Professor, Ph.D.How are the ovulated oocytes selected?WearegoingtoidentifythemechanismsofCoogenesisandfolliculogenesisusinginvitroovaryculture.AMost of oocytes degenerate in the course of development.Keywords: Oogenesis, Folliculogenesis,In vitro germ cell culture, Germ cell preservation,Animal reproductionTheobjectiveofourlaboratorymainlyfocusesonunderstandingoogenesisandfolliculogenesisinmammalianovarybygeneratinginvitroculturemethodfortheirliveimagingsystem.Backgrounds:Inmammalstheovarycontainsmorethan1millionoocytesatthemaximum.However,onlylessthan1%ofoocytesareovulatedduringreproductiveperiod,whereasmostofoocytesdegenerateinthecourseoffollicledevelopment.Thequestionsare;Howaretheovulatedoocytesselectedduringfollicledevelopment?Whatfactorsarerelatedinfolliclegrowth?Ourgoalsaretomakeclearthesequestionsandapplyournewfindingstolivestockreproductionandhumaninfertility.Research keywords?Signal transduction?Protein phosphorylation?Knock out mouse?adaptor proteinWehaveidentifiedmanyfunctionalmoleculesactingdownstreamoftheEGF(epidermalgrowthfactor)tounderstandthecomplicatednetworkofsignalingbyphospho-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.Thenumberofaminoonacidtheboundariesisindicated.primarystructureresiduesB:CLPABPKOmicewerenormalatbirthandgrewnormallyyoung.However,oldmalemicepresentedafattyphenotype,similarinmetabolicsyndrome,whenseentothatHistologicalsectionsofmouseovarycollectedat10dayafterbirth.A:Eachoocyteisenclosedinsinglefollicle,andoocytesaregrowingconcomitantlywithfolliclegrowth.B:EnlargedfigureofA.Alotofprimordialfolliclesatthesmallestfolliclestageexistasaprimordialfolliclepool(dashedcircle).50?m

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