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Cell Physiol Biochem. 2012;30(1):95-112. doi: 10.1159/000339050. Epub 2012 Jun 12.

Expression of the Slc12a1 gene in pancreatic β-cells: molecular characterization and in silico analysis.

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1
Department of Pharmacology and Toxicology, Wright State University, School of Medicine, Dayton, OH 45435, USA.

Erratum in

  • Cell Physiol Biochem. 2013;31(2-3):486. Alvarez-Leefmans, Francisco J [added].

Abstract

The solute carrier protein family 12 group A, member one (Slc12a1) and two (Slc12a2) encode several splice variants of the kidney-specific and the ubiquitous isoforms, respectively, of the bumetanide (BTD)-sensitive Na-dependent K2Cl co-transporter. The Slc12a2 co-transporter is involved in the maintenance of a high intracellular chloride concentration [Cl(-)](i) in β-cells and its inhibition with BTD blocks glucose-induced insulin secretion. In β-cells, [Cl(-)](i) plays an important role in glucose-induced depolarization and insulin secretion. Glucose promotes electrogenic efflux of Cl(-) contributing to β-cell's electrical and secretory activity. To identify the expression pattern of Slc12a1 and Slc12a2 genes in β-cells we have used RT-PCR, Western blotting and immunolocalization studies in mouse pancreatic islets, β-cell lines and rat tissues. Our results demonstrate expression of specific splice variants of Slc12a1 and Slc12a2 transcripts in β-cells i.e., variants 1 of Slc12a1 (NKCC2A) and Slc12a2 (NKCC1a). Molecular cloning and characterization of Slc12a1 variant 1 transcripts from β-cells revealed an alternative splicing event involving the 5'-UTR region. NKCC2A expression at the protein level in islets and β-cells was confirmed by immunoblotting and immunolocalization. Further, NKCC2A, NKCC1a and pro-insulin co-localized in β-cells but not in the exocrine pancreas. Therefore, our results provide for the first time evidence of NKCC2A expression in pancreatic β-cells where it may play a role in insulin secretion.

PMID:
22759959
DOI:
10.1159/000339050
[Indexed for MEDLINE]
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