Relationship between intracellular pH and metabolite concentrations during metabolic inhibition in isolated ferret heart.

1. Intracellular pH (pHi) and phosphorus metabolites were measured in isolated ferret hearts with 31P nuclear magnetic resonance (NMR). 2. The application of cyanide (to mimic hypoxia) produced a fall of the concentration of phosphocreatine ([PCr]) and a rise of those of inorganic phosphate ([Pi]) and sugar phosphates. These were accompanied by an intracellular acidosis. 3. If glycolysis was partly inhibited by prior exposure to a glucose-free solution then the application of cyanide also produced a fall of [ATP]. The acidosis was similar to that observed in the presence of glucose. 4. If glycolysis was completely inhibited by iodoacetate then the acidosis produced by subsequent addition of cyanide developed more quickly. 5. The results are reproduced by a model which incorporates lactic acid production as well as the effects of protons released and absorbed by the changes in metabolite concentrations. The results suggest that the acidosis produced by cyanide (without inhibition of glycolysis) is largely due to lactic acid production. When glycolysis is partly inhibited (glucose-free solution) the acidosis produced by cyanide is partly due to protons released by ATP breakdown. Finally, when glycolysis is entirely inhibited the acidosis is completely due to ATP breakdown. There is no need to postulate a contribution on this time scale from inhibition of pH regulating mechanisms.