HCO3？Cl？ Exchange Inactivation and Reactivation during Mouse Oocyte Meiosis Correlates with MEKMAPK-Regulated Ae2 Plasma Membrane Localization 英文参考文献.docVIP

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HCO3？Cl？ Exchange Inactivation and Reactivation during Mouse Oocyte Meiosis Correlates with MEKMAPK-Regulated Ae2 Plasma Membrane Localization 英文参考文献.doc

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HCO3？Cl？ Exchange Inactivation and Reactivation during Mouse Oocyte Meiosis Correlates with MEKMAPK-Regulated Ae2 Plasma Membrane Localization 英文参考文献

HCO32/Cl2 ExchangeInactivationandReactivation duringMouseOocyteMeiosisCorrelateswithMEK/ MAPK-RegulatedAe2PlasmaMembraneLocalization ChenxiZhou1,2,MarioTiberi1,3,4,BinhuiLiang1,3,4,SethL.Alper5,JayM.Baltz1,2,3* 1Ottawa Hospital Research Institute, Ottawa, Ontario, Canada, 2Division of Reproductive Medicine, Department of Obstetrics and Gynecology, University of Ottawa Faculty of Medicine, Ottawa, Ontario, Canada, 3Department of Cellular and Molecular Medicine, University of Ottawa Faculty of Medicine, Ottawa, Ontario, Canada, 4DepartmentsofPsychiatryandMedicine,UniversityofOttawaFacultyofMedicine,Ottawa,Ontario,Canada,5MolecularandVascularMedicineUnitandRenalDivision, BethIsraelDeaconessMedicalCenter,andDepartmentofMedicine,HarvardMedicalSchool,Boston,Massachusetts,UnitedStatesofAmerica Abstract Background: Germinal Vesicle (GV) stage mouse oocytes in first meiotic prophase exhibit highly active HCO32/Cl2 exchange—a class of transport nearly ubiquitously involved in regulation of intracellular pH and cell volume. During meiosis,however,oocyteHCO32/Cl2exchangebecomesinactivatedduringfirstmetaphase(MI),remainsinactiveinsecond metaphase (MII), and is reactivated only after egg activation. Previous work using pharmacological manipulations had indicated that activity of the MEK/MAPK signaling pathway was negatively correlated withHCO32/Cl2 exchange activity duringmeiosis.However,themechanismbywhichtheexchangerisinactivatedduringmeioticprogressionhadnotbeen determined,norhadtheroleofMEK/MAPKbeendirectlyestablished. Methodology/PrincipalFindings:ExpressionofaconstitutivelyactiveformofMEK(MAPkinasekinase),whichprevented thenormaldownregulationofMAPKaftereggactivation,alsopreventedreactivationofHCO32/Cl2exchange.Conversely, suppression of endogenous MAPK activity with dominant negative MEK activated the normally quiescent HCO32/Cl2 exchangeinmatureMIIeggs.AGFP-taggedformoftheHCO32/Cl2exchangerisoformAe2(Slc4a2)wasstronglyexpressed attheGVoocyteplasmamembrane,butmembraneloc