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Updated: Aug 1, 2026
Cell-based Calcium Assay for Medium to High Throughput Screening of TRP Channel Functions using FlexStation 3
Published on: August 17, 2011
Calcium release from pituitary secretory granules: modulation by thiols, disulfides, and dihydropyridine calcium
M Y Lorenson1, M L Cuccaro, L S Jacobs
1Department of Medicine, University of Rochester School of Medicine and Dentistry, New York 14642.
Abstract:
The distribution of calcium in isolated bovine pituitary secretory granules was studied by atomic absorption. The total granule calcium (in 26 preparations) averaged 14.5 nmol/mg protein, or 21.2 +/- 1.6% of the total pituitary homogenate calcium. Incubation of granules with KCl resulted in calcium release (78% at 15 mM and 100% at 50 mM, for example). Calcium release was also pH dependent, with greater release at acidic pH values; it was not influenced by either 500 microM strontium or 500 microM lanthanum. Release was augmented by reduced glutathione (GSH), with significant release observable at thiol levels as low as 10 microM. In addition to GSH, cysteine also stimulated release; mercaptoethanol and dithiothreitol were less potent. Interestingly, the disulfides cystine and oxidized glutathione also stimulated calcium release. Since the latter compounds are known to inhibit hormone release from granules, calcium and protein release appear to be regulated independently. A number of dihydropyridines were tested as potential blockers of calcium release from granules. Nimodipine inhibited basal calcium release at high concentrations and potently inhibited GSH-stimulated calcium release, with an apparent Ki in the 10-20 nM range; it also inhibited K(+)-stimulated release but to a lesser extent. Nimodipine, however, did not significantly influence protein or hormone release. GSH-stimulated calcium release was also inhibited by nifedipine, and this inhibition was qualitatively and quantitatively similar to that by nimodipine. Nisoldipine and nitrendipine, however, displayed no significant inhibition. In summary, it appears that the release of secretory granule calcium in vitro is independent of protein release. Thiols and some disulfides stimulate calcium release, and its inhibition by dihydropyridines suggests that granule membranes may contain specific ion channels. The role of granule calcium in the cell remains to be defined.
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