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Anoctamin 1 (Ano1) is required for glucose-induced membrane potential oscillations and insulin secretion by murine β-cells
Anions such as Cl(−) and HCO(3)(−) are well known to play an important role in glucose-stimulated insulin secretion (GSIS). In this study, we demonstrate that glucose-induced Cl(−) efflux from β-cells is mediated by the Ca(2+)-activated Cl(−) channel anoctamin 1 (Ano1). Ano1 expression in rat β-cell...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4792454/ https://www.ncbi.nlm.nih.gov/pubmed/26582426 http://dx.doi.org/10.1007/s00424-015-1758-5 |
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author | Crutzen, Raphaël Virreira, Myrna Markadieu, Nicolas Shlyonsky, Vadim Sener, Abdullah Malaisse, Willy J. Beauwens, Renaud Boom, Alain Golstein, Philippe E. |
author_facet | Crutzen, Raphaël Virreira, Myrna Markadieu, Nicolas Shlyonsky, Vadim Sener, Abdullah Malaisse, Willy J. Beauwens, Renaud Boom, Alain Golstein, Philippe E. |
author_sort | Crutzen, Raphaël |
collection | PubMed |
description | Anions such as Cl(−) and HCO(3)(−) are well known to play an important role in glucose-stimulated insulin secretion (GSIS). In this study, we demonstrate that glucose-induced Cl(−) efflux from β-cells is mediated by the Ca(2+)-activated Cl(−) channel anoctamin 1 (Ano1). Ano1 expression in rat β-cells is demonstrated by reverse transcriptase–polymerase chain reaction, western blotting, and immunohistochemistry. Typical Ano1 currents are observed in whole-cell and inside-out patches in the presence of intracellular Ca(++): at 1 μM, the Cl(−) current is outwardly rectifying, and at 2 μM, it becomes almost linear. The relative permeabilities of monovalent anions are NO(3)(−) (1.83 ± 0.10) > Br(−) (1.42 ± 0.07) > Cl(−) (1.0). A linear single-channel current–voltage relationship shows a conductance of 8.37 pS. These currents are nearly abolished by blocking Ano1 antibodies or by the inhibitors 2-(5-ethyl-4-hydroxy-6-methylpyrimidin-2-ylthio)-N-(4-(4-methoxyphenyl)thiazol-2-yl)acetamide (T-AO1) and tannic acid (TA). These inhibitors induce a strong decrease of 16.7-mM glucose-stimulated action potential rate (at least 87 % on dispersed cells) and a partial membrane repolarization with T-AO1. They abolish or strongly inhibit the GSIS increment at 8.3 mM and at 16.7 mM glucose. Blocking Ano1 antibodies also abolish the 16.7-mM GSIS increment. Combined treatment with bumetanide and acetazolamide in low Cl(−) and HCO(3)(−) media provokes a 65 % reduction in action potential (AP) amplitude and a 15-mV AP peak repolarization. Although the mechanism triggering Ano1 opening remains to be established, the present data demonstrate that Ano1 is required to sustain glucose-stimulated membrane potential oscillations and insulin secretion. |
format | Online Article Text |
id | pubmed-4792454 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-47924542016-04-09 Anoctamin 1 (Ano1) is required for glucose-induced membrane potential oscillations and insulin secretion by murine β-cells Crutzen, Raphaël Virreira, Myrna Markadieu, Nicolas Shlyonsky, Vadim Sener, Abdullah Malaisse, Willy J. Beauwens, Renaud Boom, Alain Golstein, Philippe E. Pflugers Arch Ion Channels, Receptors and Transporters Anions such as Cl(−) and HCO(3)(−) are well known to play an important role in glucose-stimulated insulin secretion (GSIS). In this study, we demonstrate that glucose-induced Cl(−) efflux from β-cells is mediated by the Ca(2+)-activated Cl(−) channel anoctamin 1 (Ano1). Ano1 expression in rat β-cells is demonstrated by reverse transcriptase–polymerase chain reaction, western blotting, and immunohistochemistry. Typical Ano1 currents are observed in whole-cell and inside-out patches in the presence of intracellular Ca(++): at 1 μM, the Cl(−) current is outwardly rectifying, and at 2 μM, it becomes almost linear. The relative permeabilities of monovalent anions are NO(3)(−) (1.83 ± 0.10) > Br(−) (1.42 ± 0.07) > Cl(−) (1.0). A linear single-channel current–voltage relationship shows a conductance of 8.37 pS. These currents are nearly abolished by blocking Ano1 antibodies or by the inhibitors 2-(5-ethyl-4-hydroxy-6-methylpyrimidin-2-ylthio)-N-(4-(4-methoxyphenyl)thiazol-2-yl)acetamide (T-AO1) and tannic acid (TA). These inhibitors induce a strong decrease of 16.7-mM glucose-stimulated action potential rate (at least 87 % on dispersed cells) and a partial membrane repolarization with T-AO1. They abolish or strongly inhibit the GSIS increment at 8.3 mM and at 16.7 mM glucose. Blocking Ano1 antibodies also abolish the 16.7-mM GSIS increment. Combined treatment with bumetanide and acetazolamide in low Cl(−) and HCO(3)(−) media provokes a 65 % reduction in action potential (AP) amplitude and a 15-mV AP peak repolarization. Although the mechanism triggering Ano1 opening remains to be established, the present data demonstrate that Ano1 is required to sustain glucose-stimulated membrane potential oscillations and insulin secretion. Springer Berlin Heidelberg 2015-11-18 2016 /pmc/articles/PMC4792454/ /pubmed/26582426 http://dx.doi.org/10.1007/s00424-015-1758-5 Text en © The Author(s) 2015 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Ion Channels, Receptors and Transporters Crutzen, Raphaël Virreira, Myrna Markadieu, Nicolas Shlyonsky, Vadim Sener, Abdullah Malaisse, Willy J. Beauwens, Renaud Boom, Alain Golstein, Philippe E. Anoctamin 1 (Ano1) is required for glucose-induced membrane potential oscillations and insulin secretion by murine β-cells |
title | Anoctamin 1 (Ano1) is required for glucose-induced membrane potential oscillations and insulin secretion by murine β-cells |
title_full | Anoctamin 1 (Ano1) is required for glucose-induced membrane potential oscillations and insulin secretion by murine β-cells |
title_fullStr | Anoctamin 1 (Ano1) is required for glucose-induced membrane potential oscillations and insulin secretion by murine β-cells |
title_full_unstemmed | Anoctamin 1 (Ano1) is required for glucose-induced membrane potential oscillations and insulin secretion by murine β-cells |
title_short | Anoctamin 1 (Ano1) is required for glucose-induced membrane potential oscillations and insulin secretion by murine β-cells |
title_sort | anoctamin 1 (ano1) is required for glucose-induced membrane potential oscillations and insulin secretion by murine β-cells |
topic | Ion Channels, Receptors and Transporters |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4792454/ https://www.ncbi.nlm.nih.gov/pubmed/26582426 http://dx.doi.org/10.1007/s00424-015-1758-5 |
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