Brain cytochrome oxidase: purification, antibody production, and immunohistochemical/histochemical correlations in the CNS
作者： RF Hevner , MT Wong-Riley
DOI: 10.1523/JNEUROSCI.09-11-03884.1989
关键词: Differential centrifugation 、 Cerebellum 、 Enzyme 、 Molecular biology 、 Affinity chromatography 、 Central nervous system 、 Biochemistry 、 Cytochrome c oxidase 、 Protein subunit 、 Cytochrome 、 Biology
摘要: Cytochrome oxidase (CO) is a mitochondrial energy-generating enzyme used in brain studies as marker of neural functional activity. The activity CO different regions, revealed histochemically, distributed nonhomogeneously but distinct patterns. Localized differences could arise from localized amount or regulation turnover number (molecular activity). To distinguish between these alternatives, we antibodies against purified calf to assess the immunohistochemical distribution (protein immunoreactivity) several regions. Calf mitochondria (synaptic and nonsynaptic populations) were isolated gray matter homogenates by differential centrifugation. was detergent extracts cytochrome c-Sepharose 4B affinity chromatography. Antisera raised rabbits. reacted specifically with CO, predominantly subunit IV, SDS immunoblots. did not react immunoblots any other proteins solubilized caudate nucleus cross-react mammalian species bovine heart CO. matched histochemical all regions tested, including monkey hippocampus mouse olfactory bulb, somatosensory (barrel) cortex, cerebellum. Thus, tissue same nonhomogeneous pattern results suggest that mechanisms exist which molecules are selectively within neurons meet local metabolic demands posed
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