Exogenous substrate utilization in Ca2+-tolerant myocytes from adult rat hearts.

Abstract

A preparation of isolated noncontracting adult rat heart myocytes can be obtained in which viability is high, yield is adequate, and the cells are not sensitive to the deleterious effects of Ca2+ at physiological levels. Myocytes are enzymatically dispersed and maintained in a medium containing amino acids and 1% bovine serum albumin at 37 degrees C. The initial viability of 77% fell to 70% after a 60-min incubation in the presence of 1.5 mM Ca2+. The initial cellular ATP content of the myocytes was 27 nmol/mg protein. Cellular ATP content fell to 21 and 17 nmol/mg protein when cells were incubated for 60 min with and without 1.5 mM Ca2+, respectively. The oxidation of glucose and lactate was increased by 50 and 30%, respectively, by the addition of Ca2+. A decrease in the oxidation rate of these two substrates was noted when the myocytes were incubated with medium containing ethyleneglycol-bis(beta-aminoethylether)-N,N'-tetraacetic acid. No changes in the rate of palmitate oxidation were observed. The significant increase in glucose and lactate oxidation due to ca2+ provides further evidence that ca2+ influences myocardial substrate utilization independent of its effects on mechanical performance of the myocardium.