Sox2 Is Essential for Formation of Trophectoderm in the Preimplantation Embryo 英文参考文献.docVIP

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Sox2 Is Essential for Formation of Trophectoderm in the Preimplantation Embryo 英文参考文献.doc

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Sox2 Is Essential for Formation of Trophectoderm in the Preimplantation Embryo 英文参考文献

Sox2IsEssentialforFormationofTrophectoderminthe PreimplantationEmbryo MariaKeramari1,JanetRazavi1,KarenA.Ingman1,ChristophPatsch2,FrankEdenhofer2,ChristopherM. Ward3,SusanJ.Kimber1* 1FacultyofLifeSciences,UniversityofManchester,Manchester,UnitedKingdom,2StemCellEngineeringGroup,InstituteofReconstructiveNeurobiology,Universityof Bonn-LifeBrainCenterandHertieFoundation,Bonn,Germany,3FacultyofMedicalandHumanSciences,UniversityofManchester,Manchester,UnitedKingdom Abstract Background: In preimplantation mammalian development the transcription factor Sox2 (SRY-related HMG-box gene 2) formsacomplexwithOct4andfunctionsinmaintenanceofself-renewalofthepluripotentinnercellmass(ICM).Previously itwasshownthatSox22/2embryosdiesoonafterimplantation.However,maternalSox2transcriptsmaymaskanearlier phenotype.WeinvestigatedwhetherSox2isinvolvedincontrollingcellfatedecisionsatanearlierstage. MethodsandFindings:WeaddressedthequestionofanearlierroleforSox2usingRNAi,whichremovesbothmaternaland embryonicSox2mRNApresentduringthepreimplantationperiod.BydepletingbothmaternalandembryonicSox2mRNAat the2-cellstageandmonitoringembryodevelopmentinvitroweshowthat,intheabsenceofSox2,embryosarrestatthe morulastageandfailtoformtrophectoderm(TE)orcavitate.Followingknock-downofSox2viathreedifferentshortinterfering RNA(siRNA)constructsin2-cellstagemouseembryos,wehaveshownthatthemajorityofembryos(76%)arrestatthemorula stageorslightlyearlierandonly18.7–21%formblastocystscomparedto76.2–83%incontrolgroups.InSox2siRNA-treated embryosexpressionofpluripotencyassociatedmarkersOct4andNanogremainedunaffected,whereasTEassociatedmarkers Tead4,Yap,Cdx2,Eomes,Fgfr2,aswellasFgf4,weredownregulatedintheabsenceofSox2.Apoptosiswasalsoincreasedin Sox2knock-downembryos.Rescueexperimentsusingcell-permeantSox2proteinresultedinincreasedblastocystformation from18.7%to62.6%andrestorationofSox2,Oct4,Cdx2andYapproteinlevelsintherescuedSox2-siRNAblastocysts. ConclusionandSignificance:WeconcludethatthefirstessentialfunctionofSox2i