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1、™METHODSINMOLECULARBIOLOGY•307LugnierLugnierSERIESEDITOR:JohnM.Walker™PhosphodiesteraseMethodsandProtocolsMETHODSINMOLECULARBIOLOGY307PhosphodiesteraseMethodsandProtocolsEditedbyClaireLugnierPharmacologieetPhysico-ChimiedesInteractionsCellulairesetMolécula
2、ires,CentreNationaldelaRechercheScientifique,UniversitéLouisPasteurdeStrasbourg,PhosphodiesterasePhosphodiesteraseFacultédePharmacie,Illkirch,FranceThemultigenicfamilyofcyclicnucleotidephosphodiesterases(PDEs)isnowknowntooffermanynewtherapeuticpossibilitie
3、sthroughtheirinfluenceoverintracellularsignaling,thoughtheirprecisecellmechanismstomodulatingthatprocessremainuncertain.InPhosphodiesteraseMethodsandProtocols,apanelofresearchleadersinthePDEfielddescribesnewconceptsandMethodsMethodstechniquesforinvestigati
4、ngtheroleofPDEsinorchestratingnormalandpathophysiologicalresponses.Presentedinstep-by-stepdetail,thesereadilyreproduciblemethodsallowthemea-surementofcyclicnucleotidevariationsinlivingcells,aswellastheirvisualizationinaspatio-temporalmanner,thelocalization
5、andcharacterizationoftheiractivitiesintissuesandlivingcells,andtheassessmentoftargetedPDEsincreatingspecifictoolsanddrugs.AdditionalchaptersandProtocolsandProtocolsdiscussthegenerationofPDE4knockoutmicetodemonstratenotonlythepotentialroleoftargetedPDEs,but
6、alsotheiruseinfurtherstudiesofthecentralroleofPDEregulationinintracellularsignalingcontrol.Thesetechniquesoffercliniciansawaytofindnewtherapiesfornumerouspathologieswhosemolecularoriginsareunknownandwhosetreatmentissymptom-atic.Alterationsofintracellularsi
7、gnalingrelatedtoPDEderegulationinsuchpathologiesasinflammation,neurodegeneration,andcancermayexplainthedifficultiesobservedintheirpreventionandtreatment.TheprotocolsfollowthesuccessfulMethodsinMolecularBiology™EditedbyEditedbyseriesformat,eachofferingstep-
8、by-steplaboratoryinstructions,anintroductionoutliningtheprinciplebehindthetechnique,listsofthenecessaryequipmentandreagents,andtipsontroubleshootingandavoidingknownpitfalls.Path-breakingandhighlypractical,Pho
