Oocyte–Targeted Deletion Reveals That Hsp90b1 Is Needed for the Completion of First Mitosis in Mouse Zygotes 英文参考文献.docVIP

Oocyte–TargetedDeletionRevealsThatHsp90b1Is NeededfortheCompletionofFirstMitosisinMouse Zygotes ChristopheAudouard1,FlorentLeMasson1,ColetteCharry1,ZihaiLi2,ElisabethS.Christians1*¤ 1CentredeBiologieduDe′veloppement, Universite′ Toulouse3-PaulSabatier,UPS,UMR5547,Toulouse,France,2DivisionofBasicSciences,HollingsCancerCenter, DepartmentofMicrobiologyandImmunology,MedicalUniversityofSouthCarolina,Charleston,SouthCarolina,UnitedStatesofAmerica Abstract Background: Hsp90b1 is an endoplasmic reticulum (ER) chaperone (also named Grp94, ERp99, gp96,Targ2, Tra-1, Tra1, Hspc4)(MGI:98817)contributingwithHspa5(alsonamedGrp78,BIP)(MGI:95835)toproteinfoldinginERcompartment. Besidesitshighproteinexpressioninmouseoocytes,littleisknownaboutHsp90b1duringthetransitionfromoocyte-to- embryo.BecausetheconstitutiveknockoutofHsp90b1isresponsibleforperi-implantationembryoniclethality,itwasnot yetknownwhetherHsp90b1isafunctionallyimportantmaternalfactor. Methodology/Findings: To circumvent embryonic lethality, we established an oocyte-specific conditional knockout line taking advantage of the more recently created floxed Hsp90b1 line (Hsp90b1flox, MGI:3700023) in combination with the transgenic mouse line expressing the cre recombinase under the control of zona pellucida 3 (ZP3) promoter (Zp3-cre, MGI:2176187). Altered expression of Hsp90b1 in growing oocytes provoked a limited, albeit significant reduction of the zona pellucida thickness but no obvious anomalies in follicular growth, meiotic maturation or fertilization. Interestingly, mutantzygotesobtainedfromoocyteslackingHsp90b1wereunabletoreachthe2-cellstage.TheyexhibitedeitheraG2/M block or, more frequently an abnormal mitotic spindle leading to developmental arrest. Despite the fact that Hspa5 displayedasimilarprofileofexpressionasHsp90b1,wefoundthatHSPA5andHSP90B1didnotfullycolocalizeinzygotes suggestingdistinctfunctionforthetwochaperones.Consequently,evenifHSPA5wasoverexpressedinHsp90b1mutant embryos, it did not compensate for H