Immunocytochemical characterization of the mitochondrially encoded ND1 subunit of complex I (NADH : ubiquinone oxidoreductase) in rat brain.
In Parkinson's disease, there is a selective defect in complex I of the electron transfer chain. To better understand complex I and its involvement in neurodegenerative disease, we raised an antibody against a conserved epitope of the human mitochondrially encoded subunit 1 of complex I (ND1). Antibodies were affinity purified and assessed by ELISA, immunoblotting, and immunocytochemistry. Immunoblots of brain homogenates from mouse, rat, and monkey brain showed a single 33-kDa band consistent with the predicted molecular mass of the protein. Subcellular fractionation showed the protein to be enriched in mitochondria. Immunocytochemistry in rat brain revealed punctate labeling in cell bodies and processes of neurons. Immunoreactively generally co-localized with subunit IV of complex IV. In striatum, ND1 immunoreactively was greatly enriched in large cholinergic neurons and neurons containing nitric oxide synthase, two cell populations that are resistant to excitotoxic and metabolic insults. In substantia nigra, many dopaminergic neurons had little ND1 immunoreactivity, which may help to explain their sensitivity to complex I inhibitors. In spinal cord, ND1 immunoreactively was enriched in motor neurons. We conclude that complex I is differentially distributed across brain regions, between neurons and glia, and between types of neurons. This antibody should provide a valuable tool for assessing complex I in normal and pathological conditions.
Pettus
E H
EH
Department of Neurology, Center for Molecular Medicine, Yerkes Regional Primate Research Center, Emory University, Atlanta, Georgia, USA.
Betarbet
R
R
Cottrell
B
B
Wallace
D C
DC
Madyastha
V
V
Greenamyre
J T
JT
D000818:Animals; D000918:Antibody Specificity; D001921:Brain / Q000201:enzymology* / Q000648:ultrastructure; D002531:Cerebellum / Q000201:enzymology; D002540:Cerebral Cortex / Q000201:enzymology; D003342:Corpus Striatum / Q000201:enzymology; D042967:Electron Transport Complex I; D004797:Enzyme-Linked Immunosorbent Assay; D005917:Globus Pallidus / Q000201:enzymology; D006624:Hippocampus / Q000201:enzymology; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008928:Mitochondria / Q000201:enzymology*; D009247:NADH, NADPH Oxidoreductases / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000201:enzymology; D013378:Substantia Nigra / Q000201:enzymology; D014018:Tissue Distribution
D009247:NADH, NADPH Oxidoreductases; D042967:Electron Transport Complex I