Metabolic studies of pyruvate- and lactate-perfused guinea pig hearts by ¹³C NMR. Determination of substrate preference by glutamate isotopomer distribution

Guinea pig hearts perfused in vitro with 99% enriched [3-¹³C]pyruvate and [3-¹³C]lactate have been examined by ¹³C NMR spectroscopy at 75.45 MHz. Resonances from the intracellular metabolites, glutamate, aspartate, alanine, citrate, malate, lactate, and acetylcarnitine are detected in the [3-¹³C]pyruvate-perfused hearts while glutamate is the only metabolite observed in the [3-¹³C]lactate-perfused hearts. Extracts obtained from individual freeze-clamped hearts run under high resolution conditions show similar distributions of metabolites indicating that the intracellular concentrations of these metabolites are indeed quite dependent upon the substrate being utilized. The excellent signal to noise in the spectra of lactate-perfused hearts allows observation of spin-spin coupling between ¹³C-enriched nuclei in the various glutamate isotopomers within the intact heart. It is shown that the steady-state distribution of glutamate isotopomers is determined by the amount of ¹²C- versus ¹³C-enriched acetyl-CoA entering the citric acid cycle and this provides the basis for a direct determination of substrate utilization by the heart in the presence of competing substrates. Similar information may be derived from the ¹³C spectrum of an extract prepared from a single [3-¹³C]pyruvate-perfused heart. Our results indicate that lactate is preferred over glucose by the guinea pig heart even in the presence of insulin.