Affiliation:
1. Department of Physiology & Biophysics, School of Medicine, University of Washington, Seattle, Washington
2. Department of Physics, Pohang University of Science & Technology, Pohang, Korea
Abstract
Pancreatic β-cells maintain glucose homeostasis by their regulated Ca2+-dependent secretion of insulin. Several cellular mechanisms control intracellular Ca2+ levels, but their relative significance in mouse β-cells is not fully known. We used photometry to measure the dynamics of cytosolic Ca2+ ([Ca2+]i) clearance after brief, depolarization-induced Ca2+ entry. Treatment with thapsigargin or cyclopiazonic acid, inhibitors of the sarco-endoplasmic reticulum Ca2+-ATPase (SERCA) pumps, nearly doubled the peak and slowed the decay of the depolarization-induced Ca2+ transients. The remaining thapsigargin-insensitive decay was slowed further by inhibition of the plasma membrane Ca2+-ATPase (PMCA) and plasma membrane Na+/Ca2+ exchanger (NCX) via alkalization of the bath solution, by adding lanthanum, or by substitution of Na+ with Li+. Mitochondrial Ca2+ uptake contributed little to clearance in thapsigargin-pretreated cells. Together, the SERCA, PMCA, and NCX transport mechanisms accounted for 89 to 97% of clearance in normal solutions. We developed a quantitative model for the dynamic role of removal mechanisms over a wide range of [Ca2+]i. According to our model, 50 to 64% of initial Ca2+ removal is via the SERCA pump, whereas the NCX contributes 21–30% of the extrusion at high [Ca2+]i, and the PMCA contributes 21–27% at low [Ca2+]i.
Publisher
American Diabetes Association
Subject
Endocrinology, Diabetes and Metabolism,Internal Medicine
Cited by
78 articles.
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