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RESEARCH ARTICLE |
1 Department of Physiological Sciences, Lund University, Sölvegatan 19, SE-223 62 Lund, Sweden
2 University of Chicago, Department of Neurobiology, Pharmacology and Physiology, 947 E. 58th Street, MC 0926, Chicago, IL 60637, USA
3 Physiologisches Institut, Universität des Saarlandes, D-66421 Homburg/Saar, Germany
*Author for correspondence (e-mail: erik.renstrom{at}mphy.lu.se)
Accepted March 3, 2001
ATP-dependent priming of the secretory granules precedes Ca2+-regulated neuroendocrine secretion, but the exact nature of this reaction is not fully established in all secretory cell types. We have further investigated this reaction in the insulin-secreting pancreatic B-cell and demonstrate that granular acidification driven by a V-type H+-ATPase in the granular membrane is a decisive step in priming. This requires simultaneous Cl- uptake through granular ClC-3 Cl- channels. Accordingly, granule acidification and priming are inhibited by agents that prevent transgranular Cl- fluxes, such as 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) and an antibody against the ClC-3 channels, but accelerated by increases in the intracellular ATP:ADP ratio or addition of hypoglycemic sulfonylureas. We suggest that this might represent an important mechanism for metabolic regulation of Ca2+-dependent exocytosis that is also likely to be operational in other secretory cell types.
Key words: ClC-3 channels, Exocytosis, Granular pH, Insulin, Sulfonylureas
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