title: Mitochondrial dysfunction in SOD1G93A-bearing astrocytes promotes motor neuron degeneration: prevention by mitochondrial-targeted antioxidants.
abstract: Mitochondrial dysfunction and oxidative stress contribute to motor neuron degeneration in amyotrophic lateral sclerosis (ALS). Recent reports indicate that astrocytes expressing the mutations of superoxide dismutase-1 (SOD1) may contribute to motor neuron injury in ALS. Here, we provide evidence that mitochondrial dysfunction in SOD1(G93A) rat astrocytes causes astrocytes to induce apoptosis of motor neurons. Mitochondria from SOD1(G93A) rat astrocytes displayed a defective respiratory function, including decreased oxygen consumption, lack of ADP-dependent respiratory control, and decreased membrane potential. Protein 3-nitrotyrosine was detected immunochemically in mitochondrial proteins from SOD1(G93A) astrocytes, suggesting that mitochondrial defects were associated with nitroxidative damage. Furthermore, superoxide radical formation in mitochondria was increased in SOD1(G93A) astrocytes. Similar defects were found in mitochondria isolated from the spinal cord of SOD1(G93A) rats, and pretreatment of animals with the spin trap 5,5-dimethyl-1-pyrroline N-oxide restored mitochondrial function, forming adducts with mitochondrial proteins in vivo. As shown previously, SOD1(G93A) astrocytes induced death of motor neurons in cocultures, compared with nontransgenic ones. This behavior was recapitulated when nontransgenic astrocytes were treated with mitochondrial inhibitors. Remarkably, motor neuron loss was prevented by preincubation of SOD1(G93A) astrocytes with antioxidants and nitric oxide synthase inhibitors. In particular, low concentrations (approximately 10 nm) of two mitochondrial-targeted antioxidants, ubiquinone and carboxy-proxyl nitroxide, each covalently coupled to a triphenylphosphonium cation (Mito-Q and Mito-CP, respectively), prevented mitochondrial dysfunction, reduced superoxide production in SOD1(G93A) astrocytes, and restored motor neuron survival. Together, our results indicate that mitochondrial dysfunction in astrocytes critically influences motor neuron survival and support the potential pharmacological utility of mitochondrial-targeted antioxidants in ALS treatment.
authors.affiliation: Departamento de Histología, and Center for Free Radical and Biomedical Research, Facultad de Medicina, Universidad de la República, 11800 Montevideo, Uruguay. pcassina@fmed.edu.uy
authors.forename: Patricia
authors.initials: P
authors.lastname: Cassina
authors.forename: Adriana
authors.initials: A
authors.lastname: Cassina
authors.forename: Mariana
authors.initials: M
authors.lastname: Pehar
authors.forename: Raquel
authors.initials: R
authors.lastname: Castellanos
authors.forename: Mandi
authors.initials: M
authors.lastname: Gandelman
authors.forename: Andrés
authors.initials: A
authors.lastname: de León
authors.forename: Kristine M
authors.initials: KM
authors.lastname: Robinson
authors.forename: Ronald P
authors.initials: RP
authors.lastname: Mason
authors.forename: Joseph S
authors.initials: JS
authors.lastname: Beckman
authors.forename: Luis
authors.initials: L
authors.lastname: Barbeito
authors.forename: Rafael
authors.initials: R
authors.lastname: Radi
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D000690:Amyotrophic Lateral Sclerosis / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control; D000818:Animals; D030801:Animals, Genetically Modified; D000975:Antioxidants / Q000008:administration & dosage*; D001253:Astrocytes / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D002478:Cells, Cultured; D016503:Drug Delivery Systems / Q000379:methods; D008928:Mitochondria / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D009046:Motor Neurons / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D009410:Nerve Degeneration / Q000201:enzymology* / Q000235:genetics / Q000517:prevention & control; D051381:Rats; D017207:Rats, Sprague-Dawley; D013482:Superoxide Dismutase / Q000235:genetics* / Q000502:physiology
chemical_list: D000975:Antioxidants; C109415:SOD1 G93A protein; D013482:Superoxide Dismutase