Inhibition of mitochondrial function affects cellular Ca2+ handling in pancreatic B-cells.
The mitochondrial inhibitors NaN(3) and carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone (FCCP) were used to study the role of mitochondria in pancreatic B-cell Ca2+ homeostasis. In glucose-stimulated B-cells NaN(3) and FCCP both increased the K(ATP) current and thus hyperpolarized the cell membrane potential, as expected for agents depleting cellular ATP. NaN(3) and FCCP stopped the glucose-induced oscillations in the cytosolic free Ca2+ concentration ([Ca2+](c)) and elicited a biphasic response. After a first rapid and transient increase, [Ca2+](c) rose in a second slow phase to a sustained level. In cells pretreated with thapsigargin the first inhibitor-induced rise in [Ca2+](c) was absent, suggesting that it may be due to Ca2+ mobilization from intracellular stores. The glucose-induced oscillations were terminated again by NaN(3) and FCCP, respectively, but the slow increase in [Ca2+](c)of the second phase was still present. A minute increase in [Ca2+](c)elicited by NaN(3) or FCCP was even visible after the removal of extracellular Ca2+, suggesting that the inhibitors also mobilize Ca2+ from mitochondria. NaN(3) and FCCP induced Ca2+ influx into B-cells treated with low glucose concentrations whose voltage-dependent Ca2+ channels are closed. Experiments with thapsigargin-preincubated cells indicate that disturbance of mitochondrial function stimulates Ca2+ influx through voltage-independent Ca2+ pathways. During the NaN(3)-induced increase in [Ca2+](c), K+-elicited depolarizations of the cells did not further augment [Ca2+](c). Evidently, this is due to a direct inhibitory effect of azide on L-type Ca2+ channels. The data demonstrate that disturbing the mitochondrial function affects cellular Ca2+ homeostasis in B-cells at several sites. Thus, it is concluded that intact mitochondrial function is a prerequisite for regular Ca2+ handling in B-cells.
Düfer
Martina
M
Pharmazeutisches Institut, Pharmakologie, Auf der Morgenstelle 8, Universität Tübingen, 72076, Germany.
Krippeit-Drews
Peter
P
Drews
Gisela
G
D000818:Animals; D001386:Azides / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D020746:Calcium Channels, L-Type / Q000187:drug effects; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D004594:Electrophysiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D066298:In Vitro Techniques; D015640:Ion Channel Gating / Q000187:drug effects; D007515:Islets of Langerhans / Q000378:metabolism*; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D051379:Mice; D008928:Mitochondria / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D019284:Thapsigargin / Q000494:pharmacology; D014475:Uncoupling Agents / Q000494:pharmacology
D001386:Azides; D020746:Calcium Channels, L-Type; D004791:Enzyme Inhibitors; D014475:Uncoupling Agents; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; D019284:Thapsigargin; D002118:Calcium