Ouabain Stimulates a Na+K+-ATPase-Mediated SFK-Activated Signalling Pathway That Regulates Tight Junction Function in the Mouse Blastocyst 英文参考文献.docVIP

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Ouabain Stimulates a Na+K+-ATPase-Mediated SFK-Activated Signalling Pathway That Regulates Tight Junction Function in the Mouse Blastocyst 英文参考文献.doc

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OuabainStimulatesaNaK-ATPase-MediatedSFK-ActivatedSignallingPathwayThatRegulatesTightJunctionFunctionintheMouseBlastocyst英文参考文献

OuabainStimulatesaNa+/K+-ATPase-MediatedSFK- ActivatedSignallingPathwayThatRegulatesTight JunctionFunctionintheMouseBlastocyst HollyGiannatselis1,2,3,4,MicheleCalder1,2,3,4,AndrewJ.Watson1,2,3,4 * 1DepartmentofObstetricsandGynaecology,TheUniversityofWesternOntario,London,Ontario,Canada,2DepartmentofPhysiologyandPharmacology,TheUniversity of Western Ontario, London, Ontario, Canada, 3Children’s Health Research Institute, London, Ontario, Canada, 4Lawson Health Research Institute, London, Ontario, Canada Abstract TheNa+/K+-ATPaseplaysapivotalroleduringpreimplantationdevelopment;itestablishesatrans-epithelialionicgradient thatfacilitatestheformationofthefluid-filledblastocystcavity,crucialforimplantationandsuccessfulpregnancy.TheNa / K -ATPaseisalsoimplicatedinregulatingtightjunctionsandcardiotonicsteroid(CTS)-inducedsignaltransductionviaSRC. + + WeinvestigatedtheexpressionofSRCfamilykinase(SFK)members,SrcandYes,duringpreimplantationdevelopmentand determinedwhetherSFKactivityisrequiredforblastocystformation.Embryoswerecollectedfollowingsuper-ovulationof CD1 or MF1 female mice. RT-PCR was used to detect SFK mRNAs encoding Src and Yes throughout preimplantation development.SRCandYESproteinwerelocalizedthroughoutpreimplantationdevelopment.Treatmentofmousemorulae withtheSFKinhibitorsPP2andSU6656for18hoursresultedinareversibleblockadeofprogressiontotheblastocyststage. Blastocyststreatedwith1023Mouabainfor2or10minutesandimmediatelyimmunostainedforphosphorylationatSRC tyr418 displayed reduced phosphorylation while in contrast blastocysts treated with 1024M displayed increased tyr418 fluorescence.SFKinhibitionincreasedandSFKactivationreducedtrophectodermtightjunctionpermeabilityinblastocysts. TheresultsdemonstratethatSFKsareexpressedduringpreimplantationdevelopmentandthatSFKactivityisrequiredfor blastocystformationandisanimportantmediatoroftrophectodermtightjunctionpermeability. Citation: Giannatselis H, Calder M, Watson AJ (2011) Ouabain Stimulates a Na+/K+-ATPase-Me