Analysis of mouse Cdh6 gene regulation by transgenesis of modified bacterial artificial chromosomes

摘要

Classiccadherinsarecelladhesionmoleculeswhoseexpressionpatternsaredynamicallymodulatedinassociationwiththeirdiversefunctionsduringmorphogenesis.Thelargesizeandcomplexityofcadherinlocihavemadeitachallengetoinvestigatetheorganizationofemcis/em-regulatorymodulesthatcontroltheirspatiotemporalpatternsofexpression.Towardsthisend,weutilizedbacterialartificialchromosomes(BACs)containingtheemCdh6/emgene,amousetypeIIclassiccadherin,tosystematicallyidentifyemcis/em-regulatorymodulesthatgovernitsexpression.ByinsertingaemlacZ/emreportergeneintotheemCdh6/emBACandgeneratingaseriesofmodifiedvariantsviahomologousrecombinationortransposoninsertionsthathavebeenexaminedintransgenicmice,weidentifiedanarrayofgenomicregionsthatcontributetospecificregulationofthegene.Theseregionsspansim;#xA0;350nbsp;kbofthelocusbetween161-kbupstreamand186-kbdownstreamoftheemCdh6/emtranscriptionstartsite.Distinctmodulesindependentlyregulatecompartmentalexpression(i.e.forebrain,hindbrainrhombomeres,andspinalcord)and/orcelllineage-specificexpressionpatterns(i.e.neuralcrestsubpopulationssuchasSchwanncells)ofemCdh6/emattheearlydevelopmentalstages.Withrespecttoregulationofexpressioninneuralcrestcells,wehavefoundthatdistinctregionscontributetodifferentaspectsofexpressionandhaveidentifiedashort79-bpregionthatisimplicatedinregulatingexpressionincellsoncetheyhaveemigratedfromtheneuraltube.TheseresultsbuildapictureofthecomplexorganizationofemCdh6cis/em-regulatorymodulesandhighlightthediverseinputsthatcontributetoitsdynamicexpressionduringearlymouseembryonicdevelopment./p/div