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Suppression of sulfonylurea- and glucose-induced insulin secretion in vitro and in vivo in mice lacking the chloride transport protein ClC-3.

Artikel i vetenskaplig tidskrift
Författare Dai-Qing Li
Xingjun Jing
Albert Salehi
Stephan C Collins
Michael B Hoppa
Anders H. Rosengren
Enming Zhang
Ingmar Lundquist
Charlotta S Olofsson
Matthias Mörgelin
Lena Eliasson
Patrik Rorsman
Erik Renström
Publicerad i Cell metabolism
Volym 10
Nummer/häfte 4
Sidor 309-15
ISSN 1932-7420
Publiceringsår 2009
Publicerad vid Institutionen för neurovetenskap och fysiologi, sektionen för fysiologi
Sidor 309-15
Språk en
Länkar dx.doi.org/10.1016/j.cmet.2009.08.0...
Ämnesord Animals, Calcium, metabolism, Chloride Channels, genetics, metabolism, Chlorides, metabolism, Cytoplasmic Granules, metabolism, Glucagon-Like Peptide 1, metabolism, Glucose, metabolism, Insulin, secretion, Insulin-Secreting Cells, cytology, drug effects, metabolism, Mice, Mice, Knockout, RNA Interference, Sulfonylurea Compounds, pharmacology
Ämneskategorier Fysiologi

Sammanfattning

Priming of insulin secretory granules for release requires intragranular acidification and depends on vesicular Cl(-)-fluxes, but the identity of the chloride transporter/ion channel involved is unknown. We tested the hypothesis that the chloride transport protein ClC-3 fulfills these actions in pancreatic beta cells. In ClC-3(-/-) mice, insulin secretion evoked by membrane depolarization (high extracellular K(+), sulfonylureas), or glucose was >60% reduced compared to WT animals. This effect was mirrored by a approximately 80% reduction in depolarization-evoked beta cell exocytosis (monitored as increases in cell capacitance) in single ClC-3(-/-) beta cells, as well as a 44% reduction in proton transport across the granule membrane. ClC-3 expression in the insulin granule was demonstrated by immunoblotting, immunostaining, and negative immuno-EM in a high-purification fraction of large dense-core vesicles (LDCVs) obtained by phogrin-EGFP labeling. The data establish the importance of granular Cl(-) fluxes in granule priming and provide direct evidence for the involvement of ClC-3 in the process.

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