Bax-Induced Apoptosis in Lebers Congenital Amaurosis A Dual Role in Rod and Cone Degeneration 英文参考文献.docVIP

Bax-InducedApoptosisinLeber’sCongenitalAmaurosis: ADualRoleinRodandConeDegeneration Se′verineHamann1,3,DanielF.Schorderet1,2,3,SandraCottet1,2 * 1IRO, Institute for Research in Ophthalmology, Sion, Switzerland, 2Department of Ophthalmology, University of Lausanne, Lausanne, Switzerland, 3School of Life Sciences,FederalInstituteofTechnology(EPFL),Lausanne,Switzerland Abstract Pathogenesis in the Rpe652/2 mouse model of Leber’s congenital amaurosis (LCA) is characterized by a slow and progressive degeneration of the rod photoreceptors. On the opposite, cones degenerate rapidly at early ages. Retinal degeneration in Rpe652/2 mice, showing a null mutation in the gene encoding the retinal pigment epithelium 65-kDa protein (Rpe65), was previously reported to depend on continuous activation of a residual transduction cascade by unligandedopsin.However,themechanismsofapoptoticsignalstriggeredbyabnormalphototransductionremainelusive. WepreviouslyreportedthatactivationofaBcl-2-dependentpathwaywasassociatedwithapoptosisofrodphotoreceptors in Rpe652/2 mice during the course of the disease. In this study we first assessed whether activation of Bcl-2-mediated apoptotic pathway was dependent on constitutive activation of the visual cascade through opsin apoprotein. We then challengedthedirectroleofpro-apoptoticBaxproteinintriggeringapoptosisofrodandconephotoreceptors. Quanti- tativePCRanalysisshowedthatincreasedexpressionofpro-apoptoticBaxanddecreasedlevelofanti-apoptoticBcl-2were restoredinRpe652/2/Gnat12/2micelackingtheGnat1geneencodingrodtransducin.Moreover,photoreceptorapoptosis waspreventedasassessedbyTUNELassay.Thesedataindicatethatabnormalactivityofopsinapoproteininducesretinal cellapoptosisthroughtheBcl-2-mediatedpathway.Followingimmunohistologicalandreal-timePCRanalyses,wefurther observed that decreased expression of rod genes in Rpe65-deficient mice was rescued in Rpe652/2/Bax2/2 mice. HistologicalandTUNELstudiesconfirmedthatrodcelldemiseandapoptosisindiseasedRpe652/2m