Combining Membrane Potential Imaging with l-Glutamate or GABA Photorelease 英文参考文献.docVIP

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Combining Membrane Potential Imaging with l-Glutamate or GABA Photorelease 英文参考文献.doc

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Combining Membrane Potential Imaging with l-Glutamate or GABA Photorelease 英文参考文献

CombiningMembranePotentialImagingwith L-GlutamateorGABAPhotorelease KasparE.Vogt1,StephanGerharz1,JeremyGraham2,MarcoCanepari1,3,4* 1DivisionofPharmacologyandNeurobiology,Biozentrum–UniversityofBasel,Basel,Switzerland,2CAIRNResearchLtd.,Faversham,UnitedKingdom,3Inserm,U836, Team3,BP170,Grenoble,France,4Universite′ JosephFourier,InstitutdesNeurosciences,BP170,Grenoble,France Abstract Combiningmembranepotentialimagingusingvoltagesensitivedyeswithphotolysisof L-glutamateorGABAallowsthe monitoringofelectricalactivityelicitedbytheneurotransmitteratdifferentsub-cellular sites.Herewedescribea simple system and some basic experimental protocols to achieve these measurements. We show how to apply the neurotransmitter and how to vary the dimension of the area of photolysis. We assess the localisation of photolysis and oftherecordedmembranepotentialchangesbydepolarisingthedendritesofcerebellarPurkinjeneuronswithL-glutamate photoreleaseusingdifferentexperimentalprotocols.WefurthershowintheapicaldendritesofCA1hippocampalpyramidal neuronshow L-glutamatephotoreleasecanbeusedtocalibratefluorescencechangesfromvoltagesensitivedyesinterms of membrane potential changes (in mV) and how GABA photorelease can be used to investigate the phenomenon of shunting inhibition. We also show how GABA photorelease can be used to measure chloride-mediated changes of membrane potential under physiological conditions originating from different regions of a neuron, providing important informationonthelocalintracellularchlorideconcentrations.Themethodandtheproofofprinciplereportedhereopen thegatewaytoavarietyofimportantapplicationswheretheadvantagesofthisapproacharenecessary. Citation:VogtKE,GerharzS,GrahamJ,CanepariM(2011)CombiningMembranePotentialImagingwith L-GlutamateorGABAPhotorelease.PLoSONE6(10): e24911.doi:10.1371/journal.pone.0024911 Editor:OlivierJacquesManzoni,InstitutNationaldelaSante′ etdelaRechercheMe′dicale,France ReceivedApril30,2011;AcceptedAugust23,2