Abstract
In situ hybridization combined with immunohistochemical techniques has been applied to study patients affected by mitochondrial myopathies with large mitochondrial (mt)DNA deletions. All patients' muscle biopsies showed ragged red fibers (RRFs) and cytochrome oxidase (COX) deficiency. Two digoxygenin-labeled, polymerase chain reaction (PCR)-amplifed DNAs were used as probes. One probe was designed to hybridize only with wild-type mtDNAs, while the other recognized both wild-type and deleted mtDNAs. Concomitant immunocytochemical analysis using antibodies against subunits II, III, (encoded by mtDNA) and IV (encoded by nuclear DNA) of COX was carried out. In our patients deleted mtDNAs are overexpressed in COX-negative RRFs, while wild-type mtDNAs are decreased in the same fibers. Immunohistochemistry studies show that COX IV is overexpressed in RRFs and that COX II and COX III subunits are still present. Deleted mtDNAs are spatially segregated in muscle fibers, where they interfere with the local population of normal mitochondrial genomes, causing a regional deficiency of the mitochondrial respiratory activity.
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Bresolin N, Moggio M, Bet L, Gallanti A, Prelle A, Nobile-Orazio E, Adobbati L, Ferrante C, Pellegrini G, Scarlato G (1986) Progressive cytochrome c oxidase deficiency in a case of Kearns-Sayre syndrome: morphological, immunological, and biochemical studies in muscle biopsies and autopsy tissues. Ann Neurol 21: 564–572
Collins S, Rudduck C, Marzuki S, Dennett X, Byrne E (1991) Mitochondrial genome distribution in histochemically cytochrome c oxidase-negative fibres in patients with a mixture of deleted and wild-type mitochondrial DNA. Biochim Biophys Acta 309: 317
Di Mauro S, Bonilla E, Zeviani M, Nagagawa M, DeVivo DC (1985) Mitochondrial myopathies. Ann Neurol 17: 521–538
Dubowitz V (1985) Muscle Biopsy: a practical approach. Balliere Tindall, London
Hammans SR, Sweeney MG, Wicks DAG, Morgan-Hughes JA, Harding AE (1992) A molecular genetic study of focal histochemical defects in mitochondrial encephalomyopathies. Brain 115: 343–365
Holt J, Harding AE, Morgan-Hughes JA (1988) Deletions of muscle mitochondrial DNA in patients with mitochondrial myopathies. Nature 331: 717–719
Lowry C, Kimmey JS, Felder S, Chi MMY, Kaiser K, Passoneau PN, Kirk KA, Lowry OH (1978) Enzyme patterns in single human muscle fibers. J Biol Chem 253: 8269–8277
Maniatis T (1982) Molecular Cloning: a laboratory manual. Cold Spring Habor Laboratory Press, Cold Spring Habor
Mita S, Schmidt B, Schon EA, DiMauro S, Bonilla E (1989) Detection of “deleted” mitochondrial genomes in cytochrome c oxidase-deficient muscle fibers of a patient with Kearns-Sayre syndrome. Med Sci 86: 9509–9513
Moraes CT, Di Mauro S, Zeviani M, Lombes A, Shanske S, Miranda AF, Nakase H, Bonilla E, Werneck LC, Servidei S, Nonaka I, Koga Y, Spiro AJ, Brownell AKW, Schotland DL, Zupanc M, De Vivo D, Schmidt B, Schon EA, Rowland LP (1989) Mitochondrial DNA deletions progressive external ophthalmoplegia and Kearns-Sayre syndrome. N Engl J Med 320: 1293–1299
Moraes CT, Ricci E, Petruzzella V, Shanske S, Di Mauro S, Schon EA, Bonilla E (1992) Molecular analysis of the muscle pathology associated with mitochondrial DNA deletions. Nature Genet 1 (5): 359–367
Nakase H, Moares CT, Rizzuto R, Lombes A, DiMauro S, Schon EA (1990) Transcription and translation of deleted mitochondrial genomes in Kearns-Sayre syndrome: implications for pathogenesis. Am J Hum Genet 46: 418–427
Olson W, Engel WK, Walsh GO, Einaugler R (1972) Oculocraniosomatic neuromuscular disease with “ragged red” fibers: histochemical and ultrastructural changes in limb muscles of a group of patients with idiopathic progressive external ophthalmoplegia. Arch Neurol 26: 193
Oldfors A, Larsson NG, Holme E, Tulinius M, Kadenbach B, Droste M (1992) Mitochondrial DNA deletion and cytochrome c oxidase deficiency in muscle fibers. J Neurol Sci 169: 177
Petty RHK, Harding AE, Morgan-Huges JA (1986) The clinical features of mitochondrial myopathies. Brain 109: 915–938
Ponzetto C, Bresolin N, Bordoni A, Moggio M, Meola G, Bet L, Prelle A, Scarlato G (1990) Kearns-Sayre syndrome: different amounts of deleted mitochondrial DNA are present in several autoptic tissues. J Neurol Sci 96: 207–210
Reichmann H (1988) Enzyme activity measured in single muscle fibers in partial COX deficiency. Neurology 38: 244–249
Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA (1988) Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239: 487–494
Schon EA, Rizzuto R, Moraes CT, Nakase H, Zeviani M, Di Mauro S (1989) A direct repeat is a hotspot for large-scale deletion of human mitochondrial DNA. Science 244: 346–349
Shoubridge EA, Karpati G, Hastings KEM (1990) Deletion mutants are functionally dominant over wild-type mitochondrial diseases. Cell 62: 43–49
Tautz D, Pfeifle C (1989) A non radioactive in situ hybridization method for the localization of specific RNAs in Drosophila embryos reveals translational control of the segmentation gene hunchback. Chromosoma 98: 81–85
Zeviani M, Moraes CT, DiMauro S, Nakase H, Bonilla E, Schon EA, Rowland LP (1988) Deletions of mitochondrial DNA in Kearns-Sayre syndrome. Neurology 38: 1339–1346
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This work was supported by the “Associazione Amici del Centro Dino Ferrari”
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Prelle, A., Fagiolari, G., Checcarelli, N. et al. Mitochondrial myopathy: correlation between oxidative defect and mitochondrial DNA deletions at single fiber level. Acta Neuropathol 87, 371–376 (1994). https://doi.org/10.1007/BF00313606
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DOI: https://doi.org/10.1007/BF00313606