title: MtDNA mutations associated with Leber's hereditary optic neuropathy: studies on cytoplasmic hybrid (cybrid) cells.
abstract: Leber's hereditary optic neuropathy (LHON) has been associated with "primary" and "secondary" mtDNA missense point mutations, and a synergistic role has been proposed for secondary mutations. No previous study has investigated the effects of LHON primary or primary plus secondary mutations on the respiratory competence of cell lines. We constructed and compared cybrid cell lines obtained from two unrelated LHON patients both carrying the common 11778/ND4 primary mutation. One of the patients also carried the 13708/ND5 and 4216/ND1 secondary mutations. The cybrid clones were evaluated for growth efficiency, oxygen consumption, complexes I, III and IV enzymatic activity and mitochondrial protein synthesis. Complex activity and mitochondrial protein synthesis were not significantly changed in cybrid clones from the patients. Oxygen consumption was significantly decreased in all clones carrying the 11778/ND4 primary mutation demonstrating its pathogenic role in impairing cell respiration. Clones also carrying the secondary mutations showed an even lower oxygen consumption and a significantly higher doubling time, suggesting that the co-presence of the secondary mutations could be relevant in further reducing the cell fitness.
authors.affiliation: Department of Neurology, University of Padova, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Vergani
authors.forename: A
authors.initials: A
authors.lastname: Martinuzzi
authors.forename: V
authors.initials: V
authors.lastname: Carelli
authors.forename: P
authors.initials: P
authors.lastname: Cortelli
authors.forename: P
authors.initials: P
authors.lastname: Montagna
authors.forename: G
authors.initials: G
authors.lastname: Schievano
authors.forename: R
authors.initials: R
authors.lastname: Carrozzo
authors.forename: C
authors.initials: C
authors.lastname: Angelini
authors.forename: E
authors.initials: E
authors.lastname: Lugaresi
mesh_terms: D000328:Adult; D001706:Biopsy; D002455:Cell Division; D002460:Cell Line; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D004272:DNA, Mitochondrial / Q000235:genetics*; D042967:Electron Transport Complex I; D003576:Electron Transport Complex IV / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000473:pathology; D006580:Genetic Carrier Screening; D006801:Humans; D006822:Hybrid Cells; D008297:Male; D009154:Mutation; D009245:NADH Dehydrogenase / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D015418:Optic Atrophies, Hereditary / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D010101:Oxygen Consumption; D010375:Pedigree; D012016:Reference Values; D012867:Skin / Q000378:metabolism* / Q000473:pathology
chemical_list: D004272:DNA, Mitochondrial; D009247:NADH, NADPH Oxidoreductases; D009245:NADH Dehydrogenase; D003576:Electron Transport Complex IV; D042967:Electron Transport Complex I