title: [Expression of a defect in the respiratory chain in cultured human cells].
abstract: Large scale deletions of mitochondrial DNA (mtDNA) or altered inter-genomic regulation in skeletal muscle have been demonstrated in patients with mitochondrial encephalomyopathies due to Cytochrome C oxidase (COx) deficiency. We have analyzed by Southern blotting and Polymerase Chain Reaction (PCR) the mtDNA in primary muscle cultures (myoblast-myotube stages and at clonal densities) and in fibrogenic subclones obtained from 9 patients with partial COx deficiency who had in their muscle biopsy a subpopulation of mtDNA showing deletions of variable size (between 2.1 and 6.5 Kb). Only in the cultures from one patient, southern analysis revealed in myoblasts and myotubes a mtDNA almost identical to that found in the original muscle biopsy and persistence of deletion in muscle cells grown at clonal densities. The deletion was detectable in fibrogenic lineage only by PCR amplification. The deleted mtDNA molecules were detectable in myogenic or fibrogenic cultures from other patients only by PCR amplification. The different amounts of deleted mtDNA in the various tissues could be due either to an unequal distribution of the altered mtDNA during embryogenesis with amplification of deleted molecules in myogenic lineage or could result from negative selection against the altered mtDNA in rapidly proliferating cells, such as fibroblasts.
authors.affiliation: Istituto di Clinica Neurologica, Centro Dino Ferrari, Università di Milano.
authors.forename: G
authors.initials: G
authors.lastname: Meola
authors.forename: G
authors.initials: G
authors.lastname: Rotondo
authors.forename: M
authors.initials: M
authors.lastname: Velicogna
authors.forename: R
authors.initials: R
authors.lastname: Toppi
authors.forename: V
authors.initials: V
authors.lastname: Sansone
authors.forename: N
authors.initials: N
authors.lastname: Bresolin
authors.forename: G
authors.initials: G
authors.lastname: Comi
authors.forename: A
authors.initials: A
authors.lastname: Bordoni
authors.forename: P
authors.initials: P
authors.lastname: Amati
authors.forename: C
authors.initials: C
authors.lastname: Ausenda
mesh_terms: D015139:Blotting, Southern; D002478:Cells, Cultured; D002872:Chromosome Deletion; D030401:Cytochrome-c Oxidase Deficiency; D004247:DNA / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics*; D004579:Electron Transport; D003576:Electron Transport Complex IV / Q000235:genetics; D005347:Fibroblasts / Q000473:pathology; D005786:Gene Expression Regulation; D006801:Humans; D008931:Mitochondria, Muscle / Q000201:enzymology*; D008956:Models, Chemical; D009124:Muscle Proteins / Q000172:deficiency / Q000235:genetics; D009132:Muscles / Q000201:enzymology / Q000473:pathology; D009245:NADH Dehydrogenase / Q000235:genetics; D009886:Ophthalmoplegia / Q000201:enzymology / Q000235:genetics*; D010086:Oxidative Phosphorylation Coupling Factors / Q000235:genetics; D016133:Polymerase Chain Reaction; D006180:Proton-Translocating ATPases / Q000235:genetics
chemical_list: D004272:DNA, Mitochondrial; D009124:Muscle Proteins; D010086:Oxidative Phosphorylation Coupling Factors; D004247:DNA; D009245:NADH Dehydrogenase; D003576:Electron Transport Complex IV; D006180:Proton-Translocating ATPases
vernacular_title: Espressione dei deficit della catena respiratoria nelle colture cellulari umane.