双顺反子表达技术在线虫神经元钙成像中的应用-论文.pdf

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1、II]I3⋯Ⅵ哪．pibb．ac．cnANovelBicistronicExpressionStrategyforNeuronalCalciumImaginginelegans冰ZHANGHai-Ning”，HUANGWen—Ming”，rANGSong”，XUTao2】”(1)KeyLaboratoryofMolecularBiophysics，MinistryofEducation,CollegeofL~eScienceandTechnology,H“azhongUniversityofScienceandTechno

2、logy,Wuhan430074，China；2)Nati0nolLaborat0ryofBiomacromolecule，InstituteofBiophysics，ChineseAcademyofSciences，Beijing100101，China)AbstractCalciumimaginghasbeenwidelyusedtomonitortheactivityofvariousneuronsineelegans．However，itisachallengetodeterminethecalciumtransi

3、entinafreelymovingwormfortworeasons．Onereasonisthechallengemensurmgtheco—expresslonofthegeneticallyencodedcalciumindicatorandreferencefluorescentproteininthesametargetneurons·Anotherreasonisthec0nlrrlonoroblemwithspectrumcross．talkbetweenthefluorescentpairusedmost

4、often：G—CaMPs(calciumindicator)andDsReds(thereferencefluorescentprotein)．Spectrumcross—talkoccasionallyintroducesartifactsinthecalciumtransientmeasuremen~-Herein,wedevelopedanovelbicistronicexpressionstrategytoensureco-expressioneficiencyandsimplifythelabelingfort

5、hetargetneurons·Moreover．wepresentedanewfluorescentproteinpairforcalciumima~ng，mKate2andG-CaMPs，whichhavelessspectrumGross．talk．Wesuccessfullytestedournewlabelingstrategyinthe￡elegansASHsensoryneuron．Weanticipatethatthisimprovedtechniquewillfacilitateneuralcircuit

6、studiesinC．elegans．Keywordsbicistronicexpression，calciumimaging，CelegansDoI：10．3724／SP．J．1206．2013．00188Animalbehaviorsarecoordinatedbyneuralcameleons，andnon—FRETproteins(alsointensity-circuitsIl1．Howthebraincontrolsbehaviorshasbeenabasedindicators)，includingG—CaM

7、P，camgaroo，andresearchhotspotfordecades．Unlikecomplexpericam．invertebratesorothervertebrates，thenematodeInbrief,bothindicatortypesaresuitableforCaenorhabditiselegansisaperfectmode1organismformeasuringthechangeincalciumintensityforalivingstudyingthisproblembecausea

8、nadulteelegansworm．However。theintensity-basedproteins，G—CaMPhemaphroditeonlyhas302neuronsandengagesinforinstance．areadvantageousbecausethedynamicvarious