Down-Regulation of ZnT8 Expression in INS-1 Rat Pancreatic Beta Cells Reduces Insulin Content and Glucose-Inducible Insulin Secretion 英文参考文献.docVIP

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Down-Regulation of ZnT8 Expression in INS-1 Rat Pancreatic Beta Cells Reduces Insulin Content and Glucose-Inducible Insulin Secretion 英文参考文献.doc

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Down-Regulation of ZnT8 Expression in INS-1 Rat Pancreatic Beta Cells Reduces Insulin Content and Glucose-Inducible Insulin Secretion 英文参考文献

Down-RegulationofZnT8ExpressioninINS-1Rat PancreaticBetaCellsReducesInsulinContentand Glucose-InducibleInsulinSecretion YiFu1,6,WeiTian1,6,EmilyB.Pratt2,5,LisaB.Dirling3,4,6,Show-LingShyng2,5,CharlesK.Meshul3,4,6 , DavidM.Cohen1,6 * 1DivisionofNephrologyHypertension,DepartmentsofMedicine,CellandDevelopmentalBiology,OregonHealthScienceUniversity,Portland,Oregon,UnitedStates ofAmerica,2DepartmentofPhysiologyandPharmacology,OregonHealthScienceUniversity,Portland,Oregon,UnitedStatesofAmerica,3DepartmentofBehavioral Neuroscience,OregonHealthScienceUniversity,Portland,Oregon,UnitedStatesofAmerica,4DepartmentofPathology,OregonHealthScienceUniversity,Portland, Oregon,UnitedStatesofAmerica,5CenterforResearchinOccupationalandEnvironmentalToxicity,OregonHealthScienceUniversity,Portland,Oregon,UnitedStates ofAmerica,6TheResearchService,PortlandV.A.MedicalCenter,Portland,Oregon,UnitedStatesofAmerica Abstract TheSLC30A8genecodesforapancreaticbeta-cell-expressedzinctransporter,ZnT8.ApolymorphismintheSLC30A8geneis associated with susceptibility to type 2 diabetes, although the molecular mechanism through which this phenotype is manifestisincompletelyunderstood.Suchpolymorphismsmayexerttheireffectviaimpactingexpressionlevelofthegene product.WeusedanshRNA-mediatedapproachtoreproduciblydownregulateZnT8mRNAexpressionby.90%intheINS- 1pancreaticbetacellline.TheZnT8-downregulatedcellsexhibiteddiminisheduptakeofexogenouszinc,asdetermined using the zinc-sensitive reporter dye, zinquin. ZnT8-downregulated cells showed reduced insulin content and decreased insulinsecretion(expressedaspercentoftotalinsulincontent)inresponsetohyperglycemicstimulus,asdeterminedby insulin immunoassay. ZnT8-depleted cells also showed fewer dense-core vesicles via electron microscopy. These data indicate that reduced ZnT8 expression in cultured pancreatic beta cells gives rise to a reduced insulin response to hyperglycemia.Inaddition,althoughweprovidenodirectevidence,thesedatasuggestthatanSLC30A8expression-level