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