JournalofVisualizedExperimentswww.jove.comCopyright©2011CreativeCommonsAttribution-NonCommercialLicenseMarch2011|49|e2533|Page1of7VideoArticleUsingWholeMountinsituHybridizationtoLinkMolecularandOrganismalBiologyNicoleL.Jacobs1,R.CraigAlbertson1,JasonR.Wiles1,21DepartmentofBiology,SyracuseUniversity2DepartmentofScienceTeaching,SyracuseUniversityCorrespondenceto:NicoleL.Jacobsatnljacobs@syr.edu,JasonR.Wilesatjwiles01@syr.eduURL:http://www.jove.com/video/2533DOI:doi:10.3791/2533Keywords:DevelopmentalBiology,Issue49,insituhybridization,genetics,development,anatomy,vertebrate,undergraduate,education,interdisciplinaryDatePublished:3/31/2011Citation:Jacobs,N.L.,Albertson,R.C.,Wiles,J.R.UsingWholeMountinsituHybridizationtoLinkMolecularandOrganismalBiology.J.Vis.Exp.(49),e2533,doi:10.3791/2533(2011).AbstractWholemountinsituhybridization(WISH)isacommontechniqueinmolecularbiologylaboratoriesusedtostudygeneexpressionthroughthelocalizationofspecificmRNAtranscriptswithinwholemountspecimen.Thistechnique(adaptedfromAlbertsonandYelick,2005)wasusedinanupperlevelundergraduateComparativeVertebrateBiologylaboratoryclassroomatSyracuseUniversity.ThefirsttwothirdsoftheComparativeVertebrateBiologylabcoursegavestudentstheopportunitytostudytheembryologyandgrossanatomyofseveralorganismsrepresentingvariouschordatetaxaprimarilyviatraditionaldissectionsandtheuseofmodels.ThefinalportionofthecourseinvolvedaninnovativeapproachtoteachinganatomythroughobservationofvertebratedevelopmentemployingmoleculartechniquesinwhichWISHwasperformedonzebrafishembryos.Aheterozygousfibroblastgrowthfactor8a(fgf8a)mutantline,ace,wasused.DuetoMendelianinheritance,aceintercrossesproducedwildtype,heterozygous,andhomozygousace/fgf8amutantsina1:2:1ratio.RNAprobeswithknownexpressionpatternsinthemidlineandindevelopinganatomicalstructuressuchastheheart,somites,tailbud,myotome,andbrainwereused.WISHwasperformedusingzebrafishatthe13somiteandprim-6stages,withstudentsperformingthestainingreactioninclass.Thestudyofzebrafishembry...
