The uptake and release of calcium by heart mitochondria

Abstract

The kinetics of uptake of Ca2+ by rat heart mitochondria were studied by a spectrophotometric method with Arsenazo III indicator. The exponential rate coefficients measured with or without added phosphate increase with the amount of Ca2+ added up to about 24μm. Evidence is given that the effect is attributable to a combination of formation of chelates at low concentrations to act as Ca2+ buffers, with co-transport of substrate to provide more respiratory fuel. The inhibitory effect of Mg2+ depends on the Ca2+ concentration, so with a constant [Mg2+] the low concentrations of Ca2+ are most inhibited, and the rate coefficients are still more Ca2+-dependent. Ca2+ uptake is slowed by local anaesthetics such as butacaine and dibucaine, and also by propranolol and palmitoyl-CoA. After an uptake, the release of Ca2+ was investigated. The spontaneous release involves an initially slow and small appearance of free Ca2+ and is followed by an auto-accelerated phase. The release is accompanied by a gradual decrease in internal ATP; it is initiated by palmitoyl-CoA (reversed by carnitine), by lysophosphatidylcholine, by Na+ salts (reversed by oligomycin) and by K+ salts added to a K+-free medium containing valinomycin. The process is probably a response to an increased energy load imposed on the mitochondria by the various conditions, which include the spontaneous action of phospholipase activated by traces of Ca2+. The problem of how much mitochondrial activity is participating in normal heart Ca2+ turnover is discussed, and experiments showing only 7–14% exchange of the mitochondrial Ca2+ occurring in vivo in 10 or 20min are reported.