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Originally published In Press as doi:10.1074/jbc.M908329199 on April 10, 2000

J. Biol. Chem., Vol. 275, Issue 26, 19978-19984, June 30, 2000
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Depletion of Intracellular Ca2+ by Caffeine and Ryanodine Induces Apoptosis of Chinese Hamster Ovary Cells Transfected with Ryanodine Receptor*

Zui PanDagger , Derek Damron§, Anna-Liisa Nieminen, Manjunatha B. BhatDagger , and Jianjie MaDagger ||

From the Departments of Dagger  Physiology & Biophysics and  Anatomy, Case Western Reserve University School of Medicine and the § Center for Anesthesiology Research, Cleveland Clinic Foundation, Cleveland, Ohio 44106

Recent studies have suggested a central role for Ca2+ in the signaling pathway of apoptosis and certain anti-apoptotic effects of Bcl-2 family of proteins have been attributed to changes in intracellular Ca2+ homeostasis. Here we report that depletion of Ca2+ from endoplasmic reticulum (ER) leads to apoptosis in Chinese hamster ovary cells. Stable expression of ryanodine receptor (RyR) in these cells enables rapid and reversible changes of both cytosolic Ca2+ and ER Ca2+ content via activation of the RyR/Ca2+ release channel by caffeine and ryanodine. Sustained depletion of the ER Ca2+ store leads to apoptosis in Chinese hamster ovary cells, whereas co-expression of Bcl-xL and RyR in these cells prevents apoptotic cell death but not necrotic cell death. The anti-apoptotic effect of Bcl-xL does not correlate with changes in either the Ca2+ release process from the ER or the capacitative Ca2+ entry through the plasma membrane. The data suggest that Bcl-xL likely prevents apoptosis of cells at a stage downstream of ER Ca2+ release and capacitative Ca2+ entry.

* This work was supported by National Institutes of Health Grant RO1-AG15556, the Charlotte Geyer Foundation, and an Established Investigatorship from the American Heart Association (to J. M.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

|| To whom correspondence should be addressed. Tel.: 216-368-2684; Fax: 216-368-1693; E-mail: jxm63@po.cwru.edu.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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