In Vivo Labeling of Mitochondrial Complex I (NADH:UbiquinoneOxidoreductase) in Rat Brain Using [3H]Dihydrorotenone

Abstract: Defects in mitochondrial energy metabolism have beenimplicated in several neurodegenerative disorders. Defective complex I(NADH:ubiquinone oxidoreductase) activity plays a key role in Leber'shereditary optic neuropathy and, possibly, Parkinson's disease, but there isno way to assess this enzyme in the living brain. We previously described anin vitro quantitative autoradiographic assay using[3H]dihydrorotenone ([3H]DHR) binding to complex I. Wehave now developed an in vivo autoradiographic assay for complex I using[3H]DHR binding after intravenous administration. In vivo[3H]DHR binding was regionally heterogeneous, and brain uptake wasrapid. Binding was enriched in neurons compared with glia, and white matterhad the lowest levels of binding. In vivo [3H]DHR binding wasmarkedly reduced by local and systemic infusion of rotenone and was enhancedby local NADH administration. There was an excellent correlation betweenregional levels of in vivo [3H]DHR binding and the in vitroactivities of complex II (succinate dehydrogenase) and complex IV (cytochromeoxidase), suggesting that the stoichiometry of these components of theelectron transport chain is relatively constant across brain regions. Theability to assay complex I in vivo should provide a valuable tool toinvestigate the status of this mitochondrial enzyme in the living brain andsuggests potential imaging techniques for complex I in humans.

[1]  D. Wallace,et al.  Immunocytochemical Characterization of the Mitochondrially Encoded ND1 Subunit of Complex I (NADH : Ubiquinone Oxidoreductase) in Rat Brain , 2000, Journal of neurochemistry.

[2]  M. Goodman,et al.  Synthesis and evaluation of a new fluorine-18 labeled rotenoid as a potential PET probe of mitochondrial complex I activity , 1999 .

[3]  J. Okun,et al.  Three Classes of Inhibitors Share a Common Binding Domain in Mitochondrial Complex I (NADH:Ubiquinone Oxidoreductase)* , 1999, The Journal of Biological Chemistry.

[4]  I. Hall,et al.  Variations in the subunit content and catalytic activity of the cytochrome c oxidase complex from different tissues and different cardiac compartments. , 1998, Biochimica et biophysica acta.

[5]  M. Ando,et al.  Marginal cells of the stria vascularis of erbils take up glucose via the facilitated transporter GLUT: application of autofluorescence , 1997, Hearing Research.

[6]  K. Frey,et al.  Intrastriatal Neurotoxin Injections Reduce in Vitro and in Vivo Binding of Radiolabeled Rotenoids to Mitochondrial Complex I , 1997, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[7]  J. Timothy Greenamyre,et al.  [3H]Dihydrorotenone Binding to NADH: Ubiquinone Reductase (Complex I) of the Electron Transport Chain: An Autoradiographic Study , 1996, The Journal of Neuroscience.

[8]  K A Frey,et al.  Synthesis and biological evaluation in mice of (2-[11C]methoxy)-6',7'-dihydrorotenol, a second generation rotenoid for marking mitochondrial complex I activity. , 1995, Nuclear medicine and biology.

[9]  M. Wong-Riley,et al.  A metabolic map of cytochrome oxidase in the rat brain: Histochemical, densitometric and biochemical studies , 1995, Neuroscience.

[10]  R. Porter,et al.  Autoradiographic study of mitochondrial complex I and glutamate receptors in the basal ganglia of rats after unilateral subthalamic lesion , 1995, Neuroscience Letters.

[11]  F. Blandini,et al.  Effect of subthalamic nucleus lesion on mitochondrial enzyme activity in rat basal ganglia , 1995, Brain Research.

[12]  J. Greene,et al.  Characterization of the Excitotoxic Potential of the Reversible Succinate Dehydrogenase Inhibitor Malonate , 1995, Journal of neurochemistry.

[13]  J. Greenamyre,et al.  Polysynaptic regulation of glutamate receptors and mitochondrial enzyme activities in the basal ganglia of rats with unilateral dopamine depletion , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[14]  R. Aggeler,et al.  Coupling between catalytic sites and the proton channel in F1F0-type ATPases. , 1994, Trends in biochemical sciences.

[15]  H. VanBrocklin,et al.  [F-18]Fluorodihydrorotenone: Synthesis and evaluation of a mitochondrial electron transport chain (ETC) complex I probe for PET , 1994 .

[16]  R. Porter,et al.  Inhibition of Succinate Dehydrogenase by Malonic Acid Produces an “Excitotoxic” Lesion in Rat Striatum , 1993, Journal of neurochemistry.

[17]  D. Hovda,et al.  Maturation of Cerebral Oxidative Metabolism in the Cat: A Cytochrome Oxidase Histochemistry Study , 1992, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[18]  D. Higgins,et al.  Quantitative Autoradiography of Dihydrorotenone Binding to Complex I of the Electron Transport Chain , 1992, Journal of neurochemistry.

[19]  R. Hevner,et al.  Coordination of ATP production and consumption in brain: Parallel regulation of cytochrome oxidase and Na+, K+-ATPase , 1992, Neuroscience Letters.

[20]  J. Casida,et al.  Interaction of 1‐Methyl‐4‐Phenylpyridinium Ion (MPP+) and Its Analogs with the Rotenone/Piericidin Binding Site of NADH Dehydrogenase , 1991, Journal of neurochemistry.

[21]  J. Parks,et al.  Evidence for a defect in NADH , 1990, Neurology.

[22]  J. Parks,et al.  Abnormalities of the electron transport chain in idiopathic parkinson's disease , 1989, Annals of neurology.

[23]  M. Wong-Riley,et al.  Brain cytochrome oxidase: purification, antibody production, and immunohistochemical/histochemical correlations in the CNS , 1989, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[24]  A. H. V. Schapira,et al.  MITOCHONDRIAL COMPLEX I DEFICIENCY IN PARKINSON'S DISEASE , 1989, The Lancet.

[25]  J. Parks,et al.  A defect in mitochondrial electron-transport activity (NADH-coenzyme Q oxidoreductase) in Leber's hereditary optic neuropathy. , 1989, The New England journal of medicine.

[26]  D. Wallace,et al.  Mitochondrial DNA mutation associated with Leber's hereditary optic neuropathy. , 1988, Science.

[27]  R. C. Collins,et al.  Distribution of Cytochrome Oxidase in Rat Brain: Studies with Diaminobenzidine Histochemistry in vitro and [14C]Cyanide Tissue Labeling in vivo , 1986, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[28]  W. Nicklas,et al.  Inhibition of NADH-linked oxidation in brain mitochondria by 1-methyl-4-phenyl-pyridine, a metabolite of the neurotoxin, 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine. , 1985, Life sciences.

[29]  C. Ragan,et al.  Photoaffinity labelling of mitochondrial NADH dehydrogenase with arylazidoamorphigenin, an analogue of rotenone. , 1984, The Biochemical journal.

[30]  J. Penney,et al.  Quantitative autoradiographic distribution of L-[3H]glutamate-binding sites in rat central nervous system , 1984, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[31]  Diana Brahams,et al.  Medicine and the Law , 1983, The Lancet.

[32]  J. Langston,et al.  Chronic Parkinsonism in humans due to a product of meperidine-analog synthesis. , 1983, Science.

[33]  Guillermo M. Alexander,et al.  Quantitative measurement of local cerebral metabolic rate for glucose utilizing tritiated 2-deoxyglucose , 1981, Brain Research.

[34]  J. Marshall,et al.  Altered succinate dehydrogenase activity of basal ganglia following damage to mesotelencephalic dopaminergic projection , 1981, Brain Research.

[35]  A. Azzi.  Cytochrome c oxidase. Towards a clarification of its structure, interactions and mechanism. , 1980, Biochimica et biophysica acta.

[36]  Louis Sokoloff,et al.  Activity‐dependent Energy Metabolism in Rat Posterior Pituitary Primarily Reflects Sodium Pump Activity , 1980, Journal of neurochemistry.

[37]  M. Wong-Riley.  Changes in the visual system of monocularly sutured or enucleated cats demonstrable with cytochrome oxidase histochemistry , 1979, Brain Research.

[38]  H. A. Padykula.  The localization of succinic dehydrogenase in tissue sections of the rat. , 1952, The American journal of anatomy.

[39]  J. Cooper,et al.  The structural organization of the mitochondrial respiratory chain , 1994 .

[40]  J. A. Volpe,et al.  5 Cytochrome c Oxidase , 1976 .