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《斑馬魚模型的心肌病相關(guān)致病基因myl的功能研究》由會員上傳分享，免費在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。
1、上海交通大學(xué)醫(yī)學(xué)院博士學(xué)位論文zebrafishasamodelorganismtoidentifyELCandRLCorthologues，respectively,andfurthermore，characterizedtheirfunctionsduringcardiogenesisandgetimplicationofELCsandRLCsinpathogenesisofcardiomyopathy．MethodsThehumanMYIApeptidesequencewasusedtosearchtheN
2、CBIdatabase，andanalysisofthenucleotideandaminoacidsequencesforthezebrafishandhumanorthologueswereperformedbyClustalWandDSGene．Whole-mountinsituhybridizationWasperformedtodetectgenesexpression．Morpholinoantisenseoligonucleotides(morpholinos)wereinjectedtoknock
3、downthegenes．MoviesofbeatingheartswererecordedusingaZeissmicroscopeequippedwithaNikoncamera．Imagesfrommovieswerethenusedtocountheartrate(HR)andmeasureshorteningfraction(SF)，enddiastolicvolume(EDV)andendsystolicvolume(ESV)．Todetectsarcomereassemblyandmeasuresa
4、rcomerelengthandcardiomyocytessize,whole-mountimmunofluorescencestainingWasperformedandimagedusinganAxioplanllZeissmicroscopeequippedwithanApotome．ThecardiomyocytesnumberwascountedafterprocessingtheimagesfromTg(cmlc2：nuDsRed)transgenicline．CelldeathWasanalyze
5、dbyTUNELassay．上海交通大學(xué)醫(yī)學(xué)院博士學(xué)位論文ResultsWeidentifiedcmlclisthemaincardiachomologueofELCandcmlc2isthemaincardiachomologueofRLCinzebrafishbybioinformaticsanalysis．Depletionofeithercmlclorcmlc2usingmorpholino—modifiedantisenseoligonucleotidesleadstoadisruptioninsarc
6、omerestructureaswellascompromisescardiacfunction，althoughthroughseeminglydistinctmechanisms．Whilemyosinstillassemblesintoarod—likestructureinbothmorphants，thesarcomerelengthislongerincmlclmorphantsbutshorterincmlc2morphantsthanthatinwildtypeembryos．Inaddition
7、，cardiomyocytesizeandnumberareincreasedupondepletionofcmlc1，resultinginalargerventricularchambervolume，whereasdepletionofcmlc2leadstoareductionincardiomyocytesizeandnumber．Thesedataindicatedthatcmlclandcmlc2arerequiredforcardiogenesisandmayhavedistinctfunctio
8、ns．ConclusionsOurdataelucidateddistinctrolesforcmlclandcmlc2duringzebrafishcardiogenesis，suggestingthatcardiomyopathiesresultingfromhumanmutationsinELCsversusRLCsmayhavedistinctpathologic
