Difference between revisions of "Kunz 1999 J Neurochem"
| Line 5: | Line 5: | ||
|year=1999 | |year=1999 | ||
|journal=J. Neurochem. | |journal=J. Neurochem. | ||
|abstract=Biochemical micromethods were used for the | |abstract=Biochemical micromethods were used for the investigation of changes in mitochondrial oxidative phosphorylation associated with cytochrome c oxidase deficiency in brain cortex from Movbr (mottled viable brindled) mice, an animal model of Menkes’ copper deficiency syndrome. Enzymatic analysis of cortex homogenates from Movbr mice showed an approximately twofold decrease | ||
investigation of changes in mitochondrial oxidative phosphorylation | in cytochrome c oxidase and a 1.4-fold decrease in NADH:cytochrome c reductase activities as compared with controls. Assessment of mitochondrial respiratory function was performed using digitonin-treated homogenates of the cortex, which exhibited the main characteristics of isolated brain mitochondria. Despite the substantial changes in respiratory chain enzyme activities, no significant differences were found in maximal pyruvate or succinate oxidation rates of brain cortex homogenates from Movbr and control mice. Inhibitor titrations were | ||
associated with cytochrome c oxidase deficiency | used to determine flux control coefficients of NADH:CoQ oxidoreductase and cytochrome c oxidase on the rate of mitochondrial respiration. Application of amobarbital to titrate the activity of NADH:CoQ oxidoreductase showed very similar flux control coefficients for control and mutant animals. Alternately, titration of respiration with azide revealed for Movbr mice significantly sharper inhibition curves than for controls, indicating a more than twofold elevated flux control coefficient of cytochrome c oxidase. Owing to the reserve capacity of respiratory chain enzymes, the reported changes in activities do not seem to affect whole-brain high-energy phosphates, as observed in a previous study using 31P NMR. | ||
in brain cortex from Movbr (mottled viable brindled) | |||
mice, an animal model of Menkes’ copper deficiency | |||
syndrome. Enzymatic analysis of cortex homogenates | |||
from Movbr mice showed an approximately twofold decrease | |||
in cytochrome c oxidase and a 1.4-fold decrease | |||
in NADH:cytochrome c reductase activities as compared | |||
with controls. Assessment of mitochondrial respiratory | |||
function was performed using digitonin-treated homogenates | |||
of the cortex, which exhibited the main characteristics | |||
of isolated brain mitochondria. Despite the substantial | |||
changes in respiratory chain enzyme activities, no | |||
significant differences were found in maximal pyruvate or | |||
succinate oxidation rates of brain cortex homogenates | |||
from Movbr and control mice. Inhibitor titrations were | |||
used to determine flux control coefficients of NADH:CoQ | |||
oxidoreductase and cytochrome c oxidase on the rate of | |||
mitochondrial respiration. Application of amobarbital to | |||
titrate the activity of NADH:CoQ oxidoreductase showed | |||
very similar flux control coefficients for control and mutant | |||
animals. Alternately, titration of respiration with azide | |||
revealed for Movbr mice significantly sharper inhibition | |||
curves than for controls, indicating a more than twofold | |||
elevated flux control coefficient of cytochrome c oxidase. | |||
Owing to the reserve capacity of respiratory chain enzymes, | |||
the reported changes in activities do not seem to | |||
affect whole-brain high-energy phosphates, as observed | |||
in a previous study using 31P NMR. | |||
|keywords=Cytochrome c oxidase deficiency, Brain mitochondria, Oxidative phosphorylation, Copper deficiency, Flux control coefficients | |keywords=Cytochrome c oxidase deficiency, Brain mitochondria, Oxidative phosphorylation, Copper deficiency, Flux control coefficients | ||
|articletype=Protocol; Manual | |articletype=Protocol; Manual | ||
| Line 42: | Line 15: | ||
|organism=Mouse | |organism=Mouse | ||
|tissues=Neurons; Brain | |tissues=Neurons; Brain | ||
|preparations=Permeabilized Cell or Tissue; Homogenate | |preparations=Isolated Mitochondria, Permeabilized Cell or Tissue; Homogenate | ||
|articletype=Protocol; Manual | |articletype=Protocol; Manual | ||
}} | }} | ||
Revision as of 16:09, 7 September 2011
| Kunz WS, Kuznetsov AV, Clark JF, Tracey I, Elger CE (1999) Metabolic consequences of the cytochrome c oxidase deficiency in brain of copper-deficient Movbr mice. J. Neurochem. 72: 1580-1585. |
Kunz WS, Kuznetsov AV, Clark JF, Tracey I, Elger CE (1999) J. Neurochem.
Abstract: Biochemical micromethods were used for the investigation of changes in mitochondrial oxidative phosphorylation associated with cytochrome c oxidase deficiency in brain cortex from Movbr (mottled viable brindled) mice, an animal model of Menkes’ copper deficiency syndrome. Enzymatic analysis of cortex homogenates from Movbr mice showed an approximately twofold decrease in cytochrome c oxidase and a 1.4-fold decrease in NADH:cytochrome c reductase activities as compared with controls. Assessment of mitochondrial respiratory function was performed using digitonin-treated homogenates of the cortex, which exhibited the main characteristics of isolated brain mitochondria. Despite the substantial changes in respiratory chain enzyme activities, no significant differences were found in maximal pyruvate or succinate oxidation rates of brain cortex homogenates from Movbr and control mice. Inhibitor titrations were used to determine flux control coefficients of NADH:CoQ oxidoreductase and cytochrome c oxidase on the rate of mitochondrial respiration. Application of amobarbital to titrate the activity of NADH:CoQ oxidoreductase showed very similar flux control coefficients for control and mutant animals. Alternately, titration of respiration with azide revealed for Movbr mice significantly sharper inhibition curves than for controls, indicating a more than twofold elevated flux control coefficient of cytochrome c oxidase. Owing to the reserve capacity of respiratory chain enzymes, the reported changes in activities do not seem to affect whole-brain high-energy phosphates, as observed in a previous study using 31P NMR. • Keywords: Cytochrome c oxidase deficiency, Brain mitochondria, Oxidative phosphorylation, Copper deficiency, Flux control coefficients
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