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Originally published In Press as doi:10.1074/jbc.M005667200 on September 19, 2000

J. Biol. Chem., Vol. 275, Issue 49, 38680-38686, December 8, 2000
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Agonist-evoked Mitochondrial Ca2+ Signals in Mouse Pancreatic Acinar Cells*

Antonio GonzálezDagger , Irene Schulz, and Andreas Schmid§

From the Department of Physiology, University of the Saarland, D-66421 Homburg/Saar, Germany

In the present study we have investigated cytosolic and mitochondrial Ca2+ signals in isolated mouse pancreatic acinar cells double-loaded with the fluorescent probes fluo-3 and rhod-2. Stimulation of pancreatic acinar cells with 500 nM acetylcholine caused release of Ca2+ from intracellular stores and produced cytosolic Ca2+ signals in form of Ca2+ waves propagating from the luminal to the basal cell pole. The increase in the cytosolic Ca2+ concentration was followed by Ca2+ uptake into mitochondria. Between onset of cytosolic and mitochondrial Ca2+ signals there was a delay of 10.7 ± 0.4 s. Ca2+ uptake into mitochondria could be inhibited with Ruthenium Red and carbonyl cyanide m-chlorophenylhydrazone, whereas 2,5-di-tert-butylhydroquinone, which inhibits sarco(endo)plasmic reticulum Ca2+ ATPases, did not prevent Ca2+ accumulation in mitochondria. Carbonyl cyanide m-chlorophenylhydrazone-induced Ca2+ release from mitochondria could only be observed after a preceding stimulation of the cell with a physiological agonist or by treatment with 2,5-di-tert-butylhydroquinone, indicating that under resting conditions mitochondria do not contain releasable Ca2+ ions. Analysis of the propagation rate of acetylcholine-induced Ca2+ waves revealed that inhibition of mitochondrial Ca2+ uptake did not accelerate spreading of cytosolic Ca2+ signals. Our experiments indicate that in the early phase of secretagogue-induced Ca2+ signals, mitochondria behave as passive Ca2+-buffering elements and do not actively suppress spreading of Ca2+ signals in pancreatic acinar cells.

* This work was supported by a Deutsche Forschungsgemeinschaft Grant (Schm 876/2-1).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.

Dagger Present address: Dept. of Physiology, University of Extremadura, Faculty of Veterinary Sciences, Avenida University, P. O. Box 643, 10071 Cáceres, Spain

§ To whom correspondence should be addressed. Tel.: 0049-6841-166454; Fax.: 0049-6841-166655; E-mail: andreas.schmid@med-rz.uni-saarland.de.

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