胸腺肽β4在小鼠卯母细胞和早期胚胎中的表达与(8)

2009;36(9)张岩等：胸腺肽鄄茁4在小鼠卵母细胞和早期胚胎中的表达与分布规律

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ExpressionandDistributionofThymosin鄄茁4in

MouseOocytesandEarlyEmbryos

ZHANGYan,LIShu-Feng,YANGCai-Rong,ZHENGKe-Jia,LINing,YANYun-Qin*

(DepartmentofAnimalCellandDevelopmentalBiology,NortheastAgriculturalUniversity,Harbin150030,China)

AbstractThymosin-茁4(thymosin-茁4,Tb4)isanimportantG-actinsequesteringfactorincells,whichcouldregulatetheactivityofG-actininvariousmicrofilamentsevents.Previously,manyresearcheshavereportedthatTb4hasbroadcellularandphysiologicalfunctions,butuntilnow,thereislackofsystematicresearchonmammaloocytesandearlyembryonicdevelopment.Tb4isasmallhydrophilicpeptide,initially,whichseparatedandpurifiedfrombovinethymuscomponent吁,withamolecularmassof5ku,andcomposeof43aminoacidresidues.Tb4moleculeishighlyconservative,whichhasbeenfoundedinavarietyoftissuesandorgansinvertebrates,andalsoexistsinpartinvertebrates.Thispeptidehasextensivebiologicalfunctions.Recentstudieshaveshownthatitcloselyrelatewithmanyphysiologicalandpathologicalactivities,suchasimmuneregulation,neurologicaldevelopment,woundhealing,inflammatoryresponse,angiogenesis,apoptosis,tumoroccurrenceandmigration.DynamiceventsofmicrofilamentsincellsalwaysaccompanywithG-actinactivity,whileTb4couldbindG-actinwitharatioof1∶1toinhibitG-actintopolymerizeintomicrofilaments.Incells,nearlyhalfofG-actinareboundwithvariousofsequesteringfactors,whicheffectsonbothG-actinactivityregulationanditsreservation,whileTb4isthechiefexecuterofG-actinsequesteringfactorfamily.Theovaries,oocytesandearlyembryosofKunmingmousewasusedtoinvestigatetheexpressionanddistributionofTb4inoocytesandearyembryos.Duringtheexperiment,theimmuno-histochemistry,immuno-fluorescence-histochemistryandRT-PCRtechniquewereutilizedaschiefstudymethods.ThedisciplineofTb4expressionanddistributioninoocytesmaturationandearlyembryonicdevelopmentwerestudiedbothinvivoandvitro.TheresultsshowedthatTb4weredifferentinexpressionandlocalizationduringmouseoocytesmaturationandearlyembryonicdevelopment.Immuno-histochemistryresultsillustratedthat,accompanywithfolliclegrowing,theexpressionamountofTb4peptideinvitrooocytesisgraduallyincreased.TherearedifferentdistributioncharacteristicsofTb4inGVstageoocytes,whichfeaturedbychromatinconfiguration,suchasdividedtheconfigurationintoNSN,pNSNandSNtype.Inotherstagesofoocytesmaturationandearlyembryonicdevelopment,Tb4couldlocatebothinnuclearandcytoplasm,andtheexpressionamountalwayschangingregularly,whicharecloselyrelatedwithcellsgrowingstatusandthecellcycle.AIODvalueofTb4wasnotsignificantlydifferentintheprocessofoocytesmaturationinvivo,butthefluorescenceintensityrapidlyincreasedform8-cellembryostoblastulastagesintheprocessofembryonicdevelopment.ThetendencyofTb4mRNAtranscriptionaresimilartothepeptideexpressioninthosestages.Specifically,inearlyimplantationblastula,Tb4peptidehighlyexpressesinimplantedlateralcellsofblastula,indicatingthatTb4canregulatethemicrofilamentsdepolymerizationinembryosimplantation,soTb4playsanimportantroleintheprocessofembryosimplantation.Theresearchconcludedthat,Tb4couldregulatemicrofilamentsevents(suchaspolymerizationanddepolymerization)andproliferationofcellsinthedevelopmentalprocessesthroughthechangesofitsexpressionandlocalization.Overall,Tb4playsanimportantroleformouseoocytesmaturation,earlyembryonicdevelopmentandembryosimplantation.

Keywordsthymosin-茁4,mouse,oocytes,embryos,microfilaments,immuno-fluorescence-histochemistry,confocal-laser-scanning-microscopeDOI:10.3724/SP.J.1206.2009.00043

*Correspondingauthor.Tel:86-451-55190846,E-mail:yanyunqin@Received:January6,2009

Accepted:March25,2009