EthanolNaltrexone Interactions at the mu-Opioid Receptor. CLSMFCS Study in Live Cells 英文参考文献.docVIP

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EthanolNaltrexone Interactions at the mu-Opioid Receptor. CLSMFCS Study in Live Cells 英文参考文献.doc

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EthanolNaltrexone Interactions at the mu-Opioid Receptor. CLSMFCS Study in Live Cells 英文参考文献

Ethanol/NaltrexoneInteractionsatthemu-Opioid Receptor.CLSM/FCSStudyinLiveCells VladanaVukojevic′1.*,YuMing1.,ClaudioD’Addario1,RudolfRigler2,3,Bjo¨rnJohansson1,LarsTerenius1 1DepartmentofClinicalNeuroscience,KarolinskaInstitutet,CMML8:01,Stockholm,Sweden,2DepartmentofMedicalBiochemistryandBiophysics,KarolinskaInstitutet, Stockholm,Sweden,3LaboratoryofBiomedicalOptics,SwissFederalInstituteofTechnology,Lausanne,Switzerland Abstract Background:Alcoholismisawidespreadchronicdisorderofcomplexaetiologywithasignificantnegativeimpactonthe individualandthesociety.Mechanismsofethanolactionarenotsufficientlywellunderstoodatthemolecularlevelandthe pharmacotherapy of alcoholism is still in its infancy. Our study focuses at the cellular and molecular level on ethanol- inducedeffectsthataremediatedthroughthemu-opioidreceptor(MOP)andontheeffectsofnaltrexone,awell-known antagonistatMOPthatisusedclinicallytopreventrelapseinalcoholism. Methodology/Principal Findings: Advanced fluorescence imaging by Confocal Laser Scanning Microscopy (CLSM) and Fluorescence Correlation Spectroscopy (FCS) are used to study ethanol effects on MOP and plasma membrane lipid dynamics in live PC12 cells. We observed that relevant concentrations of ethanol (10–40mM) alter MOP mobility and surfacedensity,andaffectthedynamicsofplasmamembranelipids.ComparedtotheactionofspecificligandsatMOP, ethanol-inducedeffectsshowcomplexkineticsandpointtoabiphasicunderlyingmechanism.Pretreatmentwithnaloxone ornaltrexoneconsiderablymitigatestheeffectsofethanol. Conclusions/Significance:WesuggestthatethanolactsbyaffectingthesortingofMOPattheplasmamembraneofPC12 cells. Naltrexone exerts opposite effects on MOP sorting at the plasma membrane, thereby countering the effects of ethanol.OurexperimentalfindingsgivenewinsightonMOP-mediatedethanolactionatthecellularandmolecularlevel. Wesuggestanewhypothesistoexplainthewellestablishedethanol-inducedincreaseintheactivityoftheendogenous opioidsystem. Citation:Vukojevic′ V,MingY,D’Addar