title: The regulation of extramitochondrial steady state free Ca2+ concentration by rat insulinoma mitochondria.
abstract: For the study of Ca2+ handling by mitochondria of an insulin secretory tissue, a method for the isolation of functionally intact insulinoma mitochondria is described. The mitochondria had a respiratory control ratio of 6.3 +/- 0.3 with succinate as a substrate. The regulation of extramitochondrial [Ca2+]o concentration by suspensions of insulinoma mitochondria was studied using Ca2+-selective minielectrodes. The mitochondria were found to maintain an ambient free Ca2+ concentration of about 0.3 and 0.9 microM in the absence or presence of Mg2+ (1 mM), respectively. The addition of Na+ resulted in a dose-dependent (half-maximal 4 mM Na+) increase in steady state [Ca2+]o. Na+ accelerated the ruthenium red-induced Ca2+ efflux, suggesting the existence of a Ca2+/2Na+ antiporter, as described in mitochondria of excitable tissues. Experiments were performed to study the effects of various agents on the steady state extramitochondrial free Ca2+. cAMP, 3-isobutyl-1-methylxanthine, and NADH were found to have no effect, whereas phosphoenolpyruvate induced a net Ca2+ efflux, the kinetic of which suggests deleterious effects on mitochondrial functions. A small decrease in pH (0.1 unit) of the incubation buffer resulted in an increase of the extramitochondrial Ca2+ steady state that was reversible upon restoration of the pH to its initial value. In conclusion, insulinoma mitochondria were able to maintain an extramitochondrial [Ca2+]o steady state in the submicromolar range that was markedly influenced by the ionic composition of the incubation medium. Thus, mitochondria may play a role in the regulation of cellular calcium homeostasis and insulin release.
mesh_terms: D007516:Adenoma, Islet Cell / Q000378:metabolism*; D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D007340:Insulinoma / Q000378:metabolism*; D007700:Kinetics; D008928:Mitochondria / Q000378:metabolism*; D009374:Neoplasms, Experimental / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012964:Sodium / Q000494:pharmacology
chemical_list: D012964:Sodium; D002118:Calcium