Disrupted Membrane Structure and Intracellular Ca2+ Signaling in Adult Skeletal Muscle with Acute Knockdown of Bin1 英文参考文献.docVIP

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Disrupted Membrane Structure and Intracellular Ca2+ Signaling in Adult Skeletal Muscle with Acute Knockdown of Bin1 英文参考文献.doc

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DisruptedMembraneStructureandIntracellularCa2SignalinginAdultSkeletalMusclewithAcuteKnockdownofBin1英文参考文献

DisruptedMembraneStructureandIntracellularCa2+ SignalinginAdultSkeletalMusclewithAcute KnockdownofBin1 AndoriaTjondrokoesoemo1,KiHoPark1,ChristopherFerrante1,ShinjiKomazaki2,SebastianLesniak1, MarcoBrotto3,Jae-KyunKo1,JingsongZhou4,NoahWeisleder1*,JianjieMa1 1DepartmentofPhysiologyandBiophysics,RobertWoodJohnsonMedicalSchool,UniversityofMedicineandDentistryofNewJersey(UMDNJ),Piscataway,NewJersey, United States of America, 2Department of Anatomy, Saitama Medical School, Saitama, Japan, 3Muscle Biology Research Group, University of Missouri, Kansas City, Missouri,UnitedStatesofAmerica,4DepartmentofMolecularBiophysicsandPhysiology,RushUniversity,Chicago,Illinois,UnitedStatesofAmerica Abstract EfficientintracellularCa2+([Ca2+]i)homeostasisinskeletalmusclerequiresintacttriadjunctionalcomplexescomprisedoft- tubuleinvaginationsofplasmamembraneandterminalcisternaeofsarcoplasmicreticulum.Bin1consistsofaspecialized BARdomainthatisassociatedwitht-tubuledevelopmentinskeletalmuscleandinvolvedintetheringthedihydropyridine receptors(DHPR)tothet-tubule.Here,weshowthatBin1isimportantforCa2+homeostasisinadultskeletalmuscle.Since systemicablationofBin1inmiceresultsinpostnatallethality,invivoelectroporationmediatedtransfectionmethodwas usedtodeliverRFP-taggedplasmidthatproducedshort–hairpin(sh)RNAtargetingBin1(shRNA-Bin1)tostudytheeffectof Bin1knockdowninadultmouseFDBskeletalmuscle.UponconfirmingthereductionofendogenousBin1expression,we showedthatshRNA-Bin1muscledisplayedswollent-tubulestructures,indicatingthatBin1isrequiredforthemaintenance ofintactmembranestructureinadultskeletalmuscle.ReducedBin1expressionledtodisruptionoft-tubulestructurethat 2+ 2+ waslinkedwithalterations tointracellularCa release.Voltage-inducedCa releasedinisolated singlemuscle fibersof shRNA-Bin1showedthatboththemeanamplitudeofCa2+currentandSRCa2+transientwerereducedwhencomparedto the shRNA-control, indicating compromised coupling between DHPR and ryanodine receptor 1. The mean frequency of 2+ osmoticst