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J Biol Chem, Vol. 274, Issue 47, 33327-33333, November 19, 1999

Regulation of Ryanodine Receptor Opening by Lumenal Ca2+ Underlies Quantal Ca2+ Release in PC12 Cells

Schuichi KoizumiDagger §, Peter LippDagger , Michael J. BerridgeDagger , and Martin D. BootmanDagger parallel

From the Dagger  Laboratory of Molecular Signalling, The Babraham Institute, Babraham Hall, Cambridge, CB2 4AT United Kingdom, the parallel  Department of Zoology, University of Cambridge, Downing Street, Cambridge, CB2 3EJ United Kingdom, and the § Division of Pharmacology, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya, Tokyo 158, Japan

Graded or "quantal" Ca2+ release from intracellular stores has been observed in various cell types following activation of either ryanodine receptors (RyR) or inositol 1,4,5-trisphosphate receptors (InsP3R). The mechanism causing the release of Ca2+ stores in direct proportion to the strength of stimulation is unresolved. We investigated the properties of quantal Ca2+ release evoked by activation of RyR in PC12 cells, and in particular whether the sensitivity of RyR to the agonist caffeine was altered by lumenal Ca2+. Quantal Ca2+ release was observed in cells stimulated with 1 to 40 mM caffeine, a range of caffeine concentrations giving a >10-fold change in lumenal Ca2+ content. The Ca2+ load of the caffeine-sensitive stores was modulated by allowing them to refill for varying times after complete discharge with maximal caffeine, or by depolarizing the cells with K+ to enhance their normal steady-state loading. The threshold for RyR activation was sensitized ~10-fold as the Ca2+ load increased from a minimal to a maximal loading. In addition, the fraction of Ca2+ released by low caffeine concentrations increased. Our data suggest that RyR are sensitive to lumenal Ca2+ over the full range of Ca2+ loads that can be achieved in an intact PC12 cell, and that changes in RyR sensitivity may be responsible for the termination of Ca2+ release underlying the quantal effect.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.


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