Cell Arrest and Cell Death in Mammalian Preimplantation Development Lessons from the Bovine Model 英文参考文献.docVIP

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Cell Arrest and Cell Death in Mammalian Preimplantation Development Lessons from the Bovine Model 英文参考文献.doc

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Cell Arrest and Cell Death in Mammalian Preimplantation Development Lessons from the Bovine Model 英文参考文献

CellArrestandCellDeathinMammalianPreimplantation Development:LessonsfromtheBovineModel SandraLeidenfrost1,MarcBoelhauve2,4,MyriamReichenbach2,TunaGu¨ngo¨r2 ,Horst-Dieter Reichenbach3,FredSinowatz1,EckhardWolf2,FelixA.Habermann1* 1InstituteofAnatomy,HistologyandEmbryology,DepartmentofVeterinarySciences,Ludwig-Maximilians-UniversityMunich,Munich,Germany,2ChairforMolecular AnimalBreedingandBiotechnology,DepartmentofVeterinarySciences,andLaboratoryforFunctionalGenomeAnalysis(LAFUGA),GeneCenter,Ludwig-Maximilians- UniversityMunich,Munich,Germany, 3BiotechnologyUnit,Institute ofAnimal Breeding, Bavarian StateInstitute for Agriculture,Poing, Germany, 4SouthWestphalia UniversityofAppliedSciences,Soest,Germany Abstract Background:Thecauses,modes,biologicalroleandprospectivesignificanceofcelldeathinpreimplantationdevelopment inhumansandothermammalsarestillpoorlyunderstood.Earlybovineembryosrepresentaveryattractiveexperimental modelfortheinvestigationofthisfundamentalandimportantissue. Methods and Findings: To obtain reference data on the temporal and spatial occurrence of cell death in early bovine embryogenesis, three-dimensionally preserved embryos of different ages and stages of development up to hatched blastocysts were examined in toto by confocal laser scanning microscopy. In parallel, transcript abundance profiles for selected apoptosis-related genes were analyzed by real-time reverse transcriptase-polymerase chain reaction. Our study documentsthatinvitroaswellasinvivo,thefirstfourcleavagecyclesarepronetoahighfailurerateincludingdifferent typesofpermanentcellcyclearrestandsubsequentnon-apoptoticblastomeredeath.Invitroproducedandinvivoderived blastocystsshowedasignificantincidenceofcelldeathintheinnercellmass(ICM),butonlyinpartwithmorphological featuresofapoptosis.Importantly,transcriptsforCASP3,CASP9,CASP8andFAS/FASLGwerenotdetectableorfoundat verylowabundances. Conclusions: In vitro and in vivo, errors and failures of the first and the next three cleavage divisio