Evaluation of mitochondrial function by measuring the heat production in state 3 and state 4 respiration

Using a microcalorimetric method, we have measured the heat production in states 3 and 4 respiration of a mitochondrial preparation from rat heart ventricle. Adenosine triphosphate production in state 3 respiration was also determined for the same preparation after heat production was measured. In Tris-buffered solution with pyruvate and malate used as substrates, the total heat production in states 3 and 4 respiration for a 10-min reaction period were about 709 and 207 mJ/mg of mitochondrial protein, respectively. 2,4-Dinitrophenol, an uncoupler of mitochondrial oxidative phosphorylation, increased the heat production in both states 3 and 4 respiration. On the other hand, antimycin A1, an inhibitor of the electron transport system, decreased the heat production in both states. When ventricle mitochondrial dysfunction was induced by the in vivo administration of a high dose of isoproterenol, heat production was increased in state 4 respiration. These results indicate that the calorimetric method can be used to evaluate the mitochondrial function.