JournalofVisualizedExperimentswww.jove.comCopyright©2011CreativeCommonsAttribution-NonCommercialLicenseNovember2011|57|e3381|Page1of6VideoArticleElectroporationofCraniofacialMesenchymeJacquelineM.Tabler1,KarenJ.Liu11DepartmentofCraniofacialDevelopment,King'sCollegeLondonURL:http://www.jove.com/video/3381DOI:doi:10.3791/3381Keywords:DevelopmentalBiology,Issue57,craniofacial,electroporation,Xenopuslaevis,frog,cartilage,mesenchymeDatePublished:11/28/2011Citation:Tabler,J.M.,Liu,K.J.ElectroporationofCraniofacialMesenchyme.J.Vis.Exp.(57),e3381,doi:10.3791/3381(2011).AbstractElectroporationisanefficientmethodofdeliveringDNAandotherchargedmacromoleculesintotissuesatprecisetimepointsandinpreciselocations.Forexample,electroporationhasbeenusedwithgreatsuccesstostudyneuralandretinaldevelopmentinXenopus,chickenandmouse1-10.However,itisimportanttonotethatinallofthesestudies,investigatorswerenottargetingsofttissues.Becauseweareinterestedincraniofacialdevelopment,weadaptedamethodtotargetfacialmesenchyme.Whenwesearchedtheliterature,wefound,tooursurprise,veryfewreportsofsuccessfulgenetransferintocartilaginoustissue.Themajorityofthesestudiesweregenetherapystudies,suchassiRNAorproteindeliveryintochondrogeniccelllines,or,animalmodelsofarthritis11-13.Inothersystems,suchaschickenormouse,electroporationoffacialmesenchymehasbeenchallenging(personalcommunications,DeptofCraniofacialDevelopment,KCL).WehypothesizedthatelectroporationintoprocartilaginousandcartilaginoustissuesinXenopusmightworkbetter.Inourstudies,weshowthatgenetransferintothefacialcartilagesoccursefficientlyatearlystages(28),whenthefacialprimordiumisstillcomprisedofsofttissuepriortocartilagedifferentiation.Xenopusisaveryaccessiblevertebratesystemforanalysisofcraniofacialdevelopment.CraniofacialstructuresaremorereadilyvisibleinXenopusthaninanyothervertebratemodel,primarilybecauseXenopusembryosarefertilizedexternally,allowinganalysesoftheearlieststages,andfacilitatingliveimagingatsinglecellresolution,aswellasreuseofthemothers...
