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

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 ex...

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Autores principales: Fu, Yi, Tian, Wei, Pratt, Emily B., Dirling, Lisa B., Shyng, Show-Ling, Meshul, Charles K., Cohen, David M.
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2009
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2682581/
https://www.ncbi.nlm.nih.gov/pubmed/19479076
http://dx.doi.org/10.1371/journal.pone.0005679
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author Fu, Yi
Tian, Wei
Pratt, Emily B.
Dirling, Lisa B.
Shyng, Show-Ling
Meshul, Charles K.
Cohen, David M.
author_facet Fu, Yi
Tian, Wei
Pratt, Emily B.
Dirling, Lisa B.
Shyng, Show-Ling
Meshul, Charles K.
Cohen, David M.
author_sort Fu, Yi
collection PubMed
description 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 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. In addition, although we provide no direct evidence, these data suggest that an SLC30A8 expression-level polymorphism could affect insulin secretion and the glycemic response in vivo.
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spelling pubmed-26825812009-05-27 Down-Regulation of ZnT8 Expression in INS-1 Rat Pancreatic Beta Cells Reduces Insulin Content and Glucose-Inducible Insulin Secretion Fu, Yi Tian, Wei Pratt, Emily B. Dirling, Lisa B. Shyng, Show-Ling Meshul, Charles K. Cohen, David M. PLoS One Research Article 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 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. In addition, although we provide no direct evidence, these data suggest that an SLC30A8 expression-level polymorphism could affect insulin secretion and the glycemic response in vivo. Public Library of Science 2009-05-25 /pmc/articles/PMC2682581/ /pubmed/19479076 http://dx.doi.org/10.1371/journal.pone.0005679 Text en This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. https://creativecommons.org/publicdomain/zero/1.0/ This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.
spellingShingle Research Article
Fu, Yi
Tian, Wei
Pratt, Emily B.
Dirling, Lisa B.
Shyng, Show-Ling
Meshul, Charles K.
Cohen, David M.
Down-Regulation of ZnT8 Expression in INS-1 Rat Pancreatic Beta Cells Reduces Insulin Content and Glucose-Inducible Insulin Secretion
title Down-Regulation of ZnT8 Expression in INS-1 Rat Pancreatic Beta Cells Reduces Insulin Content and Glucose-Inducible Insulin Secretion
title_full Down-Regulation of ZnT8 Expression in INS-1 Rat Pancreatic Beta Cells Reduces Insulin Content and Glucose-Inducible Insulin Secretion
title_fullStr Down-Regulation of ZnT8 Expression in INS-1 Rat Pancreatic Beta Cells Reduces Insulin Content and Glucose-Inducible Insulin Secretion
title_full_unstemmed Down-Regulation of ZnT8 Expression in INS-1 Rat Pancreatic Beta Cells Reduces Insulin Content and Glucose-Inducible Insulin Secretion
title_short Down-Regulation of ZnT8 Expression in INS-1 Rat Pancreatic Beta Cells Reduces Insulin Content and Glucose-Inducible Insulin Secretion
title_sort down-regulation of znt8 expression in ins-1 rat pancreatic beta cells reduces insulin content and glucose-inducible insulin secretion
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2682581/
https://www.ncbi.nlm.nih.gov/pubmed/19479076
http://dx.doi.org/10.1371/journal.pone.0005679
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