The isolated, nonworking guinea-pig heart was perfused with 14C-labelled glucose, pyruvate, and acetate. Labelled and total alanine, glutamate, aspartate, and glutamine were measured. The alanine content of the heart varied directly with the medium pyruvate concentration. The sum of the concentrations of glutamate and aspartate varied inversely with the alanine concentration. The sum of alanine, glutamate, aspartate, and glutamine in the heart was constant under most perfusion conditions, but loss into the medium increased in the presence of ouabain, low Ca2+ concentration, or malonate. The sum of the total amino acids from the heart and medium remained constant under all conditions except in the presence of malonate, when it was decreased.Labelling of amino acids from 14C-labelled substrates indicated that alanine, glutamate, and aspartate exchanged readily with their α-oxo acids. However, only 30% of the glutamine exchanged with glutamate during 1 h; this proportion varied little with perfusion conditions or the metabolic flux. The results indicate that in a closed system, most of the changes in the concentrations of amino acids are brought about by transamination.

In the perfused guinea pig heart, the rate of glucose-U-14C oxidation to 14CO2 was directly related to the external ratio [Ca2+]/[Na+]2. The metabolic stimulatory effects of ouabain were concentration-dependent, and those brought about by epinephrine were not prevented by the presence of ouabain or added Ca2+. The increased glucose utilization produced by ouabain or Ca2+ was reduced by tetrodotoxin (TTX), whereas that produced by epinephrine or reduced extracellular Na+ (100 mM) was not. It was also found that TTX inhibited the positive inotropic effects of Ca2+ and ouabain, but did not reduce the contractile force during perfusion in presence of epinephrine or at 100 mM Na+. It is concluded that (a) ouabain (or Ca2+) and epinephrine, producing additive effects, have different modes of action; (b) TTX and ouabain (or Ca2+) have antagonistic effects, probably resulting from their opposite action on calcium movements.