ATP production rate in mitochondria isolated from microsamples of human muscle

Mitochondrial ATP production (MAPR) was determined using a bioluminescence method in mitochondrial preparations of human skeletal muscle. We obtained muscle samples from 21 healthy subjects using the percutaneous muscle biopsy technique. The subjects were grouped according to their degree of physical activity, i.e., from sedentary to highly active. The MAPR for each subject was related to the mitochondrial protein content and to the activity of glutamate dehydrogenase (GDH) estimated in muscle homogenates and in isolated mitochondria. With the use of GDH as a reference base, the MAPR could be expressed per muscle mass. MAPR was determined for different individual substrates and also for the combination of pyruvate, palmitoyl-carnitine, alpha-ketoglutarate, and malate (MAPRPPKM). The MAPR observed was higher for the combination of substrates than for any individual substrate tested. The relation between mitochondrial ATP production rate and GDH activity was the same for all subjects irrespective of physical activity status, but MAPRPPKM/kg muscle varied from 6.6 +/- 1.3 mmol.min-1.kg-1 in sedentary subjects to 11.0 +/- 2.2 in highly active subjects. The present method, which uses 50 mg of muscle tissue, enables mitochondrial function to be estimated in healthy subjects or patients with mitochondrial disorders.