Mitochondrial Myopathies: Morphological Approach to Molecular Abnormalities
Cytochrome c oxidase (CCO) activity was shown by biochemical and histochemical examination to be decreased in the skeletal muscles of twelve patients with mitochondrial myopathies, especially in 8 chronic progresssive external ophthalmoplegia (CPEO) cases, which included 2 Kearns-Sayre syndrome and 6 ocular myopathy patients. In 4 MELAS patients, NADH cytochrome c reductase activity was decreased. Immunocytochemical examination, using anti-CCO, anti-complex I and III rabbit sera revealed that CCO was stained more weakly in the muscle fibers of one of the CPEO patients than in those of the control.
Immuno-electron microscopic examination of CCO, complex I and III, using a method of gold labeling, was also performed. Extensive labeling by gold particles, representing the localization of respiratory enzymes, could be seen in close vicinity to the cristae and inner mitochondrial membrane of normal shaped mitochondria. The concentration of gold particles was markedly decreased in one of the CPEO patients.
To detect the localization of mitochondrial DNA or mRNA, in situ hybridization was performed on human biopsied muscles using a 35S labeled mitochondrial DNA probe. The wide distribution of autoradiographic grains for mRNA over the sarcoplasm of all muscle fibers was correlated with the distribution of immuno-stained mitochondria.
Southern blotting revealed large deletions of mitochondrial DNA in six of the patients with CPEO. In one of these patients, in situ hybridization showed a marked decrease in density of autoradiographic grains in muscle fibers.
KeywordsGold Particle Mitochondrial Myopathy Chronic Progressive External Ophthalmoplegia Myoclonus Epilepsy Biopsy Muscle Specimen
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