down-regulation of znt8 expression in ins-1 rat pancreatic beta cells reduces insulin content and glucose-inducible insulin secretionznt8表达下调ins-1鼠胰腺β细胞减少胰岛素含量和glucose-inducible胰岛素分泌.pdfVIP

Down-Regulation of ZnT8 Expression in INS-1 Rat Pancreatic Beta Cells Reduces Insulin Content and Glucose-Inducible Insulin Secretion Yi Fu1,6, Wei Tian1,6, Emily B. Pratt2,5, Lisa B. Dirling3,4,6, Show-Ling Shyng2,5, Charles K. Meshul3,4,6, David M. Cohen1,6* 1 Division of Nephrology Hypertension, Departments of Medicine, Cell and Developmental Biology, Oregon Health Science University, Portland, Oregon, United States of America, 2 Department of Physiology and Pharmacology, Oregon Health Science University, Portland, Oregon, United States of America, 3 Department of Behavioral Neuroscience, Oregon Health Science University, Portland, Oregon, United States of America, 4 Department of Pathology, Oregon Health Science University, Portland, Oregon, United States of America, 5 Center for Research in Occupational and Environmental Toxicity, Oregon Health Science University, Portland, Oregon, United States of America, 6 The Research Service, Portland V.A. Medical Center, Portland, Oregon, United States of America Abstract The SLC30A8 gene codes for a pancreatic beta-cell-expressed zinc transporter, ZnT8. A polymorphism in the SLC30A8 gene is associated with susceptibility to type 2 diabetes, although the molecular mechanism through which this phenotype is manifest is incompletely understood. Such polymorphisms may exert their effect via impacting expression level of the gene product. We used an shRNA-mediated approach to reproducibly downregulate ZnT8 mRNA expression by .90% in the INS- 1 pancreatic beta cell line. The ZnT8-downregulated cells exhibited diminished uptake of exogenous zinc, as determined using the zinc-sensitive reporter dye, zinquin. ZnT8-downregulated cells showed reduced insulin content and decreased insulin secretion (expressed as percent of total insulin content) in response to hyperglycemic stimulus, as determined by insulin immunoassay. ZnT8-depleted cells al