title: Manganese superoxide dismutase levels are elevated in a proportion of amyotrophic lateral sclerosis patient cell lines.
abstract: The most frequent genetic causes of amyotrophic lateral sclerosis (ALS) determined so far are mutations occurring in the gene for copper/zinc superoxide dismutase (CuZnSOD). The mechanism may involve inappropriate formation of hyroxyl radicals, peroxynitrite or malfunctioning of the SOD protein. We hypothesized that undiscovered genetic causes of sporadically occurring amyotrophic lateral sclerosis might be found in the mechanisms that create and destroy oxygen free radicals within the cell. After determining that there were no CuZnSOD mutations present, we measured superoxide production from mitochondria and manganese superoxide dismutase (MnSOD), glutathione peroxidase, NFkappaB, Bcl-2 and Bax by immunoblot. Of the ten sporadic patients we tested we found three patients with significantly increased concentrations of MnSOD. These patients also had lower levels of superoxide production from mitochondria and decreased expression of Bcl-2. No mutations were found in the cDNA sequence of either MnSOD in any of the sporadic patients. A patient with a CuZnSOD mutation (G82R) used as a positive control showed none of these abnormalities. The patients displaying the MnSOD aberrations showed no specific distinguishing features. This result suggests that the cause of ALS in a subgroup of ALS patients (30%) is genetic in origin and can be identified by these markers. The alteration in MnSOD and Bcl-2 are likely epiphenomena resulting from the primary genetic defect. It suggests also that the oxygen free radicals are part of the cause in this subgroup and that dysregulation of MnSOD or increased endogenous superoxide production might be responsible.
authors.affiliation: Metabolism Research Programme, Research Institute, Hospital for Sick Children, University of Toronto, Canada.
authors.forename: G
authors.initials: G
authors.lastname: McEachern
authors.forename: S
authors.initials: S
authors.lastname: Kassovska-Bratinova
authors.forename: S
authors.initials: S
authors.lastname: Raha
authors.forename: M A
authors.initials: MA
authors.lastname: Tarnopolsky
authors.forename: J
authors.initials: J
authors.lastname: Turnbull
authors.forename: J
authors.initials: J
authors.lastname: Bourgeois
authors.forename: B
authors.initials: B
authors.lastname: Robinson
mesh_terms: D000328:Adult; D000368:Aged; D019943:Amino Acid Substitution / Q000235:genetics; D000690:Amyotrophic Lateral Sclerosis / Q000201:enzymology* / Q000235:genetics / Q000473:pathology*; D015153:Blotting, Western; D002478:Cells, Cultured; D004252:DNA Mutational Analysis; D005347:Fibroblasts / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D005979:Glutathione Peroxidase / Q000378:metabolism; D006801:Humans; D008875:Middle Aged; D008928:Mitochondria / Q000201:enzymology / Q000378:metabolism; D008970:Molecular Weight; D016328:NF-kappa B / Q000032:analysis; D010641:Phenotype; D011518:Proto-Oncogene Proteins / Q000032:analysis; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000032:analysis; D013482:Superoxide Dismutase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism; D015854:Up-Regulation; D051028:bcl-2-Associated X Protein
chemical_list: C494826:BAX protein, human; D016328:NF-kappa B; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D013481:Superoxides; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase