Enzymatic Relay Mechanism Stimulates Cyclic GMP Synthesis in Rod Photoresponse Biochemical and Physiological Study in Guanylyl Cyclase Activating Protein 1 Knockout Mice 英文参考文献.docVIP

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Enzymatic Relay Mechanism Stimulates Cyclic GMP Synthesis in Rod Photoresponse Biochemical and Physiological Study in Guanylyl Cyclase Activating Protein 1 Knockout Mice 英文参考文献.doc

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Enzymatic Relay Mechanism Stimulates Cyclic GMP Synthesis in Rod Photoresponse Biochemical and Physiological Study in Guanylyl Cyclase Activating Protein 1 Knockout Mice 英文参考文献

EnzymaticRelayMechanismStimulatesCyclicGMP SynthesisinRodPhotoresponse:Biochemicaland PhysiologicalStudyinGuanylylCyclaseActivating Protein1KnockoutMice ClintL.Makino1,Xiao-HongWen1.,ElenaV.Olshevskaya2.,IgorV.Peshenko2.,AndreyB.Savchenko2, AlexanderM.Dizhoor2* 1DepartmentofOphthalmology,MassachusettsEyeandEarInfirmaryandHarvardMedicalSchool,Boston,Massachusetts,UnitedStatesofAmerica,2Departmentof BasicSciencesandPennsylvaniaCollegeofOptometry,SalusUniversity,ElkinsPark,Pennsylvania,UnitedStatesofAmerica Abstract Regulation of cGMP synthesis by retinal membrane guanylyl cyclase isozymes (RetGC1 and RetGC2) in rod and cone photoreceptorsbycalcium-sensitiveguanylylcyclaseactivatingproteins(GCAP1andGCAP2)isoneofthekeymolecular mechanismsaffectingtheresponsetolightandisinvolvedincongenitalretinaldiseases.Theobjectiveofthisstudywasto identifythephysiologicalsequenceofeventsunderlyingRetGCactivationinvivo,bystudyingtheelectrophysiologicaland biochemical properties of mouse rods in a new genetic model lacking GCAP1. The GCAP12/2 retinas expressed normal levels of RetGC isozymes and other phototransduction proteins, with the exception of GCAP2, whose expression was 2+ elevated in a compensatory fashion. RetGC activity in GCAP12/2 retinas became more sensitive to Ca and slightly increased. The bright flash response in electroretinogram (ERG) recordings recovered quickly in GCAP12/2, as well as in RetGC12/2 GCAP1 2/2,andRetGC22/2GCAP12/2hybridrods,indicatingthatGCAP2activatesbothRetGCisozymesinvivo. IndividualGCAP12/2rodresponsesvariedinsizeandshape,likelyreflectingvariableendogenousGCAP2levelsbetween different cells, but single-photon response (SPR) amplitude and time-to-peak were typically increased, while recovery kineticsremainedfasterthaninwildtype.RecoveryfrombrightflashesinGCAP12/2 wasprominentlybiphasic,because rare,aberrantSPRsproducingtheslowertailcomponentweremagnified.Thesedataprovidestrongphysiologicalevidence thatrodphotoresponserecoveryisshapedbytheseq