JournalofVisualizedExperimentswww.jove.comCopyright©2011CreativeCommonsAttribution-NonCommercialLicenseAugust2011|54|e3024|Page1of6VideoArticleMouseinUteroElectroporation:ControlledSpatiotemporalGeneTransfectionAsukaMatsui1,AyaC.Yoshida1,MayumiKubota1,MasaharuOgawa1,TomomiShimogori11LabforMolecularMechanismsofThalamusDevelopment,RIKENBrainScienceInstituteCorrespondenceto:TomomiShimogoriattshimogori@brain.riken.jpURL:http://www.jove.com/video/3024DOI:doi:10.3791/3024Keywords:Neuroscience,Issue54,Inutero,electroporation,transfection,CNS,geneexpression,gainoffunction,lossoffunction,neuron,axonDatePublished:8/15/2011Citation:Matsui,A.,Yoshida,A.C.,Kubota,M.,Ogawa,M.,Shimogori,T.MouseinUteroElectroporation:ControlledSpatiotemporalGeneTransfection.J.Vis.Exp.(54),e3024,doi:10.3791/3024(2011).AbstractInordertounderstandthefunctionofgenesexpressedinspecificregionofthedevelopingbrain,includingsignalingmoleculesandaxonguidancemolecules,localgenetransferorknock-outisrequired.Genetargetingknock-inorknock-outintolocalregionsispossibletoperformwithcombinationwithaspecificCREline,whichislaborious,costly,andtimeconsuming.Therefore,asimpletransfectionmethod,aninuteroelectroporationtechnique,whichcanbeperformedwithshorttime,willbehandytotestthepossiblefunctionofcandidategenespriortothegenerationoftransgenicanimals1,2.Inadditiontothis,inuteroelectroporationtargetsareasofthebrainwherenospecificCRElineexists,andwilllimitembryoniclethality3,4.Here,wepresentamethodofinuteroelectroporationcombiningtwodifferenttypesofelectrodesforsimpleandconvenientgenetransferintotargetareasofthedevelopingbrain.First,auniqueholdingmethodofembryosusinganopticfiberopticlightcablewillmakesmallembryos(fromE9.5)visiblefortargetedDNAsolutioninjectionintoventriclesandneedletypeelectrodesinsertiontothetargetedbrainarea5,6.Thepatterningofthebrainsuchascorticalareaoccuratearlyembryonicstage,therefore,theseearlyelectroporationfromE9.5makeabigcontributiontounderstandentireareapatterningevent.Second,thepreciseshapeo...
