Faculty Collaboration Database

Brain cytochrome oxidase: purification, antibody production, and immunohistochemical/histochemical correlations in the CNS. J Neurosci 1989 Nov;9(11):3884-98

Date

11/01/1989

Pubmed ID

2555458

Pubmed Central ID

PMC6569932

DOI

10.1523/JNEUROSCI.09-11-03884.1989

Scopus ID

2-s2.0-0024355783 (requires institutional sign-in at Scopus site)   96 Citations

Abstract

Cytochrome oxidase (CO) is a mitochondrial energy-generating enzyme used in brain studies as a marker of neural functional activity. The activity of CO in different brain regions, revealed histochemically, is distributed nonhomogeneously but in distinct patterns. Localized differences in CO activity could arise from localized differences in enzyme amount or from localized regulation of enzyme turnover number (molecular activity). To distinguish between these alternatives, we used antibodies against purified calf brain CO to assess the immunohistochemical distribution of CO amount (protein immunoreactivity) in several brain regions. Calf brain mitochondria (synaptic and nonsynaptic populations) were isolated from gray matter homogenates by differential centrifugation. CO was purified from detergent extracts of the mitochondria by cytochrome c-Sepharose 4B affinity chromatography. Antisera against the purified CO were raised in rabbits. The antibodies reacted specifically with CO, predominantly subunit IV, in SDS immunoblots. The antibodies did not react in SDS immunoblots with any other proteins solubilized from mitochondria or caudate nucleus but did cross-react with brain CO from other mammalian species and with bovine heart CO. The immunohistochemical distribution of CO amount matched the histochemical distribution of CO activity in all regions tested, including the monkey hippocampus and the mouse olfactory bulb, somatosensory (barrel) cortex, and cerebellum. Thus, the amount of CO in neural tissue is distributed in the same nonhomogeneous pattern as the histochemical activity of CO. The results suggest that mechanisms exist by which CO molecules are selectively distributed within neurons to meet local metabolic demands posed by neural functional activity.

Author List

Hevner RF, Wong-Riley MT

MESH terms used to index this publication - Major topics in bold

Animals
Brain
Cattle
Cell Fractionation
Chromatography, Affinity
Electron Transport Complex IV
Enzyme-Linked Immunosorbent Assay
Immunoblotting
Immunoenzyme Techniques
Immunoglobulin G
Kinetics
Microscopy, Electron
Mitochondria
Mitochondria, Heart
Molecular Weight
Synapses