The neuronal K+Cl− co-transporter 2 (Slc12a5) modulates insulin secretion
Abstract Intracellular chloride concentration ([Cl−]i) in pancreatic β-cells is kept above electrochemical equilibrium due to the predominant functional presence of Cl− loaders such as the Na+K+2Cl− co-transporter 1 (Slc12a2) over Cl−extruders of unidentified nature. Using molecular cloning, RT-PCR,...
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doaj-d1c00f15f43e41669bcde19b13071f672020-12-08T02:30:02ZengNature Publishing GroupScientific Reports2045-23222017-05-017111410.1038/s41598-017-01814-0The neuronal K+Cl− co-transporter 2 (Slc12a5) modulates insulin secretionShams Kursan0Timothy S. McMillen1Pavani Beesetty2Eduardo Dias-Junior3Mohammed M. Almutairi4Abu A. Sajib5J. Ashot Kozak6Lydia Aguilar-Bryan7Mauricio Di Fulvio8Department of Pharmacology and Toxicology, Wright State University, School of MedicinePacific Northwest Diabetes Research InstituteDepartment of Neuroscience, Cell Biology and Physiology, Wright State University, School of MedicineDepartment of Pharmacology and Toxicology, Wright State University, School of MedicineDepartment of Pharmacology and Toxicology, Wright State University, School of MedicineDepartment of Genetic Engineering and Biotechnology, University of DhakaDepartment of Neuroscience, Cell Biology and Physiology, Wright State University, School of MedicinePacific Northwest Diabetes Research InstituteDepartment of Pharmacology and Toxicology, Wright State University, School of MedicineAbstract Intracellular chloride concentration ([Cl−]i) in pancreatic β-cells is kept above electrochemical equilibrium due to the predominant functional presence of Cl− loaders such as the Na+K+2Cl− co-transporter 1 (Slc12a2) over Cl−extruders of unidentified nature. Using molecular cloning, RT-PCR, Western blotting, immunolocalization and in vitro functional assays, we establish that the “neuron-specific” K+Cl− co-transporter 2 (KCC2, Slc12a5) is expressed in several endocrine cells of the pancreatic islet, including glucagon secreting α-cells, but particularly in insulin-secreting β-cells, where we provide evidence for its role in the insulin secretory response. Three KCC2 splice variants were identified: the formerly described KCC2a and KCC2b along with a novel one lacking exon 25 (KCC2a-S25). This new variant is undetectable in brain or spinal cord, the only and most abundant known sources of KCC2. Inhibition of KCC2 activity in clonal MIN6 β-cells increases basal and glucose-stimulated insulin secretion and Ca2+ uptake in the presence of glibenclamide, an inhibitor of the ATP-dependent potassium (KATP)-channels, thus suggesting a possible mechanism underlying KCC2-dependent insulin release. We propose that the long-time considered “neuron-specific” KCC2 co-transporter is expressed in pancreatic islet β-cells where it modulates Ca2+-dependent insulin secretion.https://doi.org/10.1038/s41598-017-01814-0 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Shams Kursan Timothy S. McMillen Pavani Beesetty Eduardo Dias-Junior Mohammed M. Almutairi Abu A. Sajib J. Ashot Kozak Lydia Aguilar-Bryan Mauricio Di Fulvio |
spellingShingle |
Shams Kursan Timothy S. McMillen Pavani Beesetty Eduardo Dias-Junior Mohammed M. Almutairi Abu A. Sajib J. Ashot Kozak Lydia Aguilar-Bryan Mauricio Di Fulvio The neuronal K+Cl− co-transporter 2 (Slc12a5) modulates insulin secretion Scientific Reports |
author_facet |
Shams Kursan Timothy S. McMillen Pavani Beesetty Eduardo Dias-Junior Mohammed M. Almutairi Abu A. Sajib J. Ashot Kozak Lydia Aguilar-Bryan Mauricio Di Fulvio |
author_sort |
Shams Kursan |
title |
The neuronal K+Cl− co-transporter 2 (Slc12a5) modulates insulin secretion |
title_short |
The neuronal K+Cl− co-transporter 2 (Slc12a5) modulates insulin secretion |
title_full |
The neuronal K+Cl− co-transporter 2 (Slc12a5) modulates insulin secretion |
title_fullStr |
The neuronal K+Cl− co-transporter 2 (Slc12a5) modulates insulin secretion |
title_full_unstemmed |
The neuronal K+Cl− co-transporter 2 (Slc12a5) modulates insulin secretion |
title_sort |
neuronal k+cl− co-transporter 2 (slc12a5) modulates insulin secretion |
publisher |
Nature Publishing Group |
series |
Scientific Reports |
issn |
2045-2322 |
publishDate |
2017-05-01 |
description |
Abstract Intracellular chloride concentration ([Cl−]i) in pancreatic β-cells is kept above electrochemical equilibrium due to the predominant functional presence of Cl− loaders such as the Na+K+2Cl− co-transporter 1 (Slc12a2) over Cl−extruders of unidentified nature. Using molecular cloning, RT-PCR, Western blotting, immunolocalization and in vitro functional assays, we establish that the “neuron-specific” K+Cl− co-transporter 2 (KCC2, Slc12a5) is expressed in several endocrine cells of the pancreatic islet, including glucagon secreting α-cells, but particularly in insulin-secreting β-cells, where we provide evidence for its role in the insulin secretory response. Three KCC2 splice variants were identified: the formerly described KCC2a and KCC2b along with a novel one lacking exon 25 (KCC2a-S25). This new variant is undetectable in brain or spinal cord, the only and most abundant known sources of KCC2. Inhibition of KCC2 activity in clonal MIN6 β-cells increases basal and glucose-stimulated insulin secretion and Ca2+ uptake in the presence of glibenclamide, an inhibitor of the ATP-dependent potassium (KATP)-channels, thus suggesting a possible mechanism underlying KCC2-dependent insulin release. We propose that the long-time considered “neuron-specific” KCC2 co-transporter is expressed in pancreatic islet β-cells where it modulates Ca2+-dependent insulin secretion. |
url |
https://doi.org/10.1038/s41598-017-01814-0 |
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