Optical Silencing of C. elegans Cells with Arch Proton Pump 英文参考文献.docVIP

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Optical Silencing of C. elegans Cells with Arch Proton Pump 英文参考文献.doc

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Optical Silencing of C. elegans Cells with Arch Proton Pump 英文参考文献

OpticalSilencingofC.elegansCellswithArchProton Pump AyakoOkazaki,YukiSudo,ShinTakagi* DivisionofBiologicalScience,GraduateSchoolofScience,NagoyaUniversity,Nagoya,Japan Abstract Background: Optogenetic techniques using light-driven ion channels or ion pumps for controlling excitable cells have greatlyfacilitatedtheinvestigationofnervoussystemsinvivo.Amodelorganism,C.elegans,withitssmalltransparentbody andwell-characterizedneuralcircuits,isespeciallysuitableforoptogeneticanalyses. Methodology/Principal Findings: We describe the application of archaerhodopsin-3 (Arch), a recently reported optical neuronalsilencer,toC.elegans.Arch::GFPexpressedeitherinallneuronsorbodywallmusclesoftheentirebodybymeans of transgenes were localized, at least partially, to the cell membrane without adverse effects, and caused locomotory paralysisofwormswhenilluminatedbygreenlight(550nm).Pan-neuronalexpressionofArchendowedwormswithquick and sustained responsiveness to such light. Worms reliably responded to repeated periods of illumination and non- illumination, and remained paralyzed under continuous illumination for 30 seconds. Worms expressing Arch in different subsetsofmotorneuronsexhibiteddistinctdefectsinthelocomotorybehaviorundergreenlight:selectivesilencingofA- type motorneurons affected backward movement while silencing ofB-type motorneurons affected forward movement more severely. Our experiments using a heat-shock-mediated induction system also indicate that Arch becomes fully functionalonly12hoursafterinductionandremainsfunctionalformorethan24hour. Conclusions/Sgnificance:Archcanbeusedforsilencingneuronsandmuscles,andmaybeausefulalternativetocurrently widelyusedhalorhodopsin(NpHR)inoptogeneticstudiesofC.elegans. Citation: Okazaki A, Sudo Y, Takagi S (2012) Optical Silencing of C. elegans Cells with Arch Proton Pump. PLoS ONE 7(5): e35370. doi:10.1371/ journal.pone.0035370 Editor:HitoshiOkazawa,TokyoMedicalandDentalUniversity,Japan ReceivedNovember29,2011;AcceptedMarch16,2012;P