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Papers In Press, published online ahead of print September 19, 2000
Department of Physiology, University of the Saarland, Homburg/Saar 66421
Corresponding Author: andreas.schmid{at}med-rz.uni-saarland.de
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 (ACh) 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 (CCCP), whereas 2,5-di-tert-butylhydroquinone (tBHQ), which inhibits sarco(endo)plasmic reticulum Ca2+ ATPases did not prevent Ca2+ accumulation in mitochondria. CCCP-induced Ca2+ release from mitochondria could only be observed after a preceding stimulation of the cell with a physiological agonist or by treatment with tBHQ, indicating that under resting conditions mitochondria do not contain releasable Ca2+ ions. Analysis of the propagation rate of ACh-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.
J. Biol. Chem, 10.1074/jbc.M005667200
Submitted on June 28, 2000
Revised on September 5, 2000
Accepted on September 18, 2000
Agonist-evoked mitochondrial Ca2+ signals in mouse pancreatic acinar cells
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