Early changes of muscle mitochondria in Duchenne dystrophy. Partition and activity of mitochondrial enzymes in fractionated muscle of unaffected boys and adults and patients.
(1) Biopsies from the gastrocnemius muscle of patients with Duchenne dystrophy were partitioned into a myofibrillar plus nuclear fraction, a mitochondrial fraction and a supernatant fraction. The fractions were assayed for mitochondrial enzymes and protein, in order to obtain information about the integrity of mitochondrial structure and function. Muscles from boys and adults without neuromuscular disease were treated likewise. (2) In adults, muscle possesses a significantly higher specific activity (on protein basis) of monoamine oxidase and rotenone-insenitive NADH-cytochrome c reductase (RINCR) than in boys. In childhood, monoamine oxidase activity increases with age. At the age of 5 yr, the specific activity is 50% of the adult value. RINCR activity is constant in childhood. With adolescence it increases from 20 +/- 2 (SEM) to 35 +/- 6 mumoles cytochrome c reduced per min per g protein, and it remains at this level. Palmitoyl-CoA synthetase activity remains constant with age. (3) In Duchenne dystrophy the extractable protein content from muscle is decreased to 75%. The specific activities of the matrix enzymes propionyl-CoA carboxylase and glutamate dehydrogenase are 1.8 and 2.8 times increased, the inner membrane enzyme cytochrome c oxidase is 2.8 times increased, the inner membrane enzyme cytochrome c oxidase is 2.8 times increased. Of the outer membrane enzymes RINCR is 2.0 times increased, while palmitoyl-CoA synthetase is not changed in acitivity. In Duchenne dystrophy monoamine oxidase activity also increases with age. In part this may be due to mitochondria from adipose tissue and macrophages, which are increasingly present in older patients. The specific activities of enzymes with a predominant cytosolic localisation, creatine kinase and adenylate kinase, are increased by a factor of 1.5 and 1.7. (4) The subcellular distribution of the studied enzymes in human skeletal muscle was found to be similar as in animal studies. In mitochondrial fractions from Duchenne patients the recoveries of the following enzymes are decreased: glutamate dehydrogenase (from  25 to 9%), creatine kinase (1.1-0.66%), adenylate kinase (0.44-0.22%), hexokinase (7.1-2.7%), monoamine oxidase (36-21%), RINCR (30-17%), and palmitoyl-CoA synthetase (40-21%). The recoveries of last 3 mitochondrial outer membrane enzymes in the supernatant fractions are correspondingly increased. These results indicate an increased fragility of the mitochondrial membranes in dystrophic muscles. (5) The reported changes are clearly evident in a one-year-old patient, which indicates that the mitochondria are involved early in the disease process.
D000263:Adenylate Kinase / Q000378:metabolism; D000293:Adolescent; D002262:Carboxy-Lyases / Q000378:metabolism; D002458:Cell Fractionation; D002648:Child; D002675:Child, Preschool; D003066:Coenzyme A Ligases / Q000378:metabolism; D003402:Creatine Kinase / Q000378:metabolism; D003574:Cytochrome c Group; D003576:Electron Transport Complex IV / Q000378:metabolism; D005969:Glutamate Dehydrogenase / Q000378:metabolism; D006593:Hexokinase / Q000378:metabolism; D006801:Humans; D007223:Infant; D008297:Male; D043586:Methylmalonyl-CoA Decarboxylase; D008931:Mitochondria, Muscle / Q000201:enzymology / Q000378:metabolism*; D008995:Monoamine Oxidase / Q000378:metabolism; D009124:Muscle Proteins / Q000032:analysis; D009136:Muscular Dystrophies / Q000378:metabolism*; D009245:NADH Dehydrogenase / Q000378:metabolism; D010171:Palmitoyl Coenzyme A; D011422:Propionates / Q000378:metabolism
D003574:Cytochrome c Group; D009124:Muscle Proteins; D011422:Propionates; D010171:Palmitoyl Coenzyme A; D005969:Glutamate Dehydrogenase; D008995:Monoamine Oxidase; D009245:NADH Dehydrogenase; D003576:Electron Transport Complex IV; D006593:Hexokinase; D003402:Creatine Kinase; D000263:Adenylate Kinase; D002262:Carboxy-Lyases; D003066:Coenzyme A Ligases; D043586:Methylmalonyl-CoA Decarboxylase