资源描述:
《1. Spatial Parkin Translocation and Degradation of Damaged Mitochondria via Mitophagy in Live Cortical Neurons 姚溢洲 》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、CurrentBiology22,545552,March20,2012ª2012ElsevierLtdAllrightsreservedDOI10.1016/j.cub.2012.02.005ReportSpatialParkinTranslocationandDegradationofDamagedMitochondriaviaMitophagyinLiveCorticalNeuronsQianCai,1,2,*HeshamMostafaZakaria,1,3AnthonySimone,1vehicledimethylsulfoxide
2、(DMSO)asacontrol,10mMandZu-HangSheng1,*carbonylcyanidem-chlorophenylhydrazone(CCCP;aDcm1SynapticFunctionsSection,PorterNeuroscienceResearchdissipatingreagent),or10mMCCCPwithlysosomalinhibitorsCenter,NationalInstituteofNeurologicalDisordersand(LIs;10mMpepstatinAand10mME64D)
3、.AlthoughYFP-ParkinStroke,NationalInstitutesofHealth,Room2B-215,wasdiffuseinthecytosolofDMSO-treatedcontrolneurons35ConventDrive,Bethesda,MD20892-3706,USA(n=435),itredistributedtomitochondriain26.67%64.46%2DepartmentofCellBiologyandNeuroscience,Rutgersofneurons(n=420)treat
4、edwithCCCPandin55.87%6University,604AllisonRoad,Piscataway,NJ08854-6999,USA6.57%ofneuronstreatedwithCCCP/LIs(n=570)(Figures3HowardHughesMedicalInstitute,NationalInstitutesof1Aand1B).TreatmentwithCCCP/LIsdoubledthepercentageHealthResearchScholarsProgram,1CloisterCourt,ofneu
5、ronswithParkintranslocationrelativetoCCCPaloneBuilding60,Bethesda,MD20814,USA(p<0.001),suggestingthatlysosomaldegradationcapacityhasasignificantimpactontheclearanceofParkin-targetedmitochondriaviamitophagyinneurons.Second,wecoimmu-Summarynostainedsimilarlytreatedneuronswith
6、antibodiesagainstneuronalmarkerMAP2andmitochondrialmarkersTOM20Mitochondriaareessentialforneuronalsurvivalandfunc-(anoutermembraneprotein)orcytochromec(adynamicinter-tion.Properdegradationofagedanddamagedmitochondriamembranespaceprotein).YFP-Parkinwasrecruitedtomito-throug
7、hmitophagyisakeycellularpathwayformitochon-chondrialabeledwithTOM20orcytochromec(Figure1C)indrialqualitycontrol.RecentstudieshaveindicatedthatCCCP-treatedneurons,butnotinDMSOcontrols.ToexaminePINK1/Parkin-mediatedpathwaysensuremitochondrialParkintranslocationkinetics,weima
8、gedneuronsatvariousintegrityandfunction[1–8].TranslocationofParkintotimepointsduringCCCPt