Increased mitochondrial superoxide generation in neurons from trisomy 16 mice: a model of Down's syndrome.
Increased neuronal cell death in neurodegenerative diseases has been suggested to result from an increased mitochondrial generation of radical oxygen species (ROS). To test this hypothesis, we investigated superoxide formation in cultured hippocampal neurons from diploid and trisomy 16 mice (Ts16), a model of Down's syndrome. Microflurometric techniques were used to measure superoxide-induced oxidation rate of hydroethidine (HEt) to ethidium and reduced nicotinamide adenine dinucleotide (NADH) and reduced nicotinamide adenine dinucleotide phosphate (NADPH) autofluorescence signal to monitor changes in neuronal energy metabolism. We found an increase in superoxide formation by more than 50% in Ts16 neurons in comparison with diploid control neurons. In the presence of the mitochondrial respiratory chain complex I inhibitor rotenone superoxide production was blocked in diploid neurons, but the increased superoxide generation in Ts16 neurons remained. Uncoupling of mitochondrial oxidative phosphorylation using carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) caused irreversible deficiency in the energy metabolism, monitored by NAD(P)H autofluorescence in Ts16 neurons, but not in diploid control neurons. These results suggest an increased basal generation of superoxide in Ts16 neurons, probably caused by a deficient complex I of mitochondrial electron transport chain, which leads to an impaired mitochondrial energy metabolism and finally neuronal cell death.
Institut für Physiologie der Charité, Humboldt Universität Berlin, Germany. sebastian.schuchmann@charite.de
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Heinemann
D000818:Animals; D000968:Antimycin A / Q000494:pharmacology; D016923:Cell Death; D002478:Cells, Cultured; D002874:Chromosome Mapping; D003486:Cyanides / Q000494:pharmacology; D004171:Diploidy; D004195:Disease Models, Animal; D004314:Down Syndrome / Q000235:genetics / Q000473:pathology / Q000503:physiopathology*; D042967:Electron Transport Complex I; D014450:Electron Transport Complex III / Q000378:metabolism; D003576:Electron Transport Complex IV / Q000378:metabolism; D005314:Embryonic and Fetal Development / Q000235:genetics; D006624:Hippocampus / Q000473:pathology / Q000503:physiopathology; D006801:Humans; D051379:Mice; D008817:Mice, Mutant Strains; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology*; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000473:pathology / Q000502:physiology*; D017382:Reactive Oxygen Species / Q000502:physiology; D012402:Rotenone / Q000494:pharmacology; D013481:Superoxides / Q000378:metabolism*; D014314:Trisomy
D003486:Cyanides; D017382:Reactive Oxygen Species; D012402:Rotenone; D013481:Superoxides; D000968:Antimycin A; D009247:NADH, NADPH Oxidoreductases; D003576:Electron Transport Complex IV; D042967:Electron Transport Complex I; D014450:Electron Transport Complex III