Use of mitochondrial inhibitors to demonstrate that cytochrome oxidase near-infrared spectroscopy can measure mitochondrial dysfunction noninvasively in the brain.
The use of near-infrared spectroscopy to measure noninvasively changes in the redox state of cerebral cytochrome oxidase in vivo is controversial. We therefore tested these measurements using a multiwavelength detector in the neonatal pig brain. Exchange transfusion with perfluorocarbons revealed that the spectrum of cytochrome oxidase in the near-infrared was identical in the neonatal pig, the adult rat, and in the purified enzyme. Under normoxic conditions, the neonatal pig brain contained 15 micromol/L deoxyhemoglobin, 29 micromol/L oxyhemoglobin, and 1.2 micromol/L oxidized cytochrome oxidase. The mitochondrial inhibitor cyanide was used to determine whether redox changes in cytochrome oxidase could be detected in the presence of the larger cerebral hemoglobin concentration. Addition of cyanide induced full reduction of cytochrome oxidase in both blooded and bloodless animals. In the blooded animals, subsequent anoxia caused large changes in hemoglobin oxygenation and concentration but did not affect the cytochrome oxidase near-infrared signal. Simultaneous blood oxygenation level-dependent magnetic resonance imaging measurements showed a good correlation with near-infrared measurements of deoxyhemoglobin concentration. Possible interference in the near-infrared measurements from light scattering changes was discounted by simultaneous measurements of the optical pathlength using the cerebral water absorbance as a standard chromophore. We conclude that, under these conditions, near-infrared spectroscopy can accurately measure changes in the cerebral cytochrome oxidase redox state.
Cooper
C E
CE
Department of Biological Sciences, University of Essex, Colchester, UK.
Cope
M
M
Springett
R
R
Amess
P N
PN
Penrice
J
J
Tyszczuk
L
L
Punwani
S
S
Ordidge
R
R
Wyatt
J
J
Delpy
D T
DT
D000818:Animals; D001921:Brain / Q000201:enzymology* / Q000648:ultrastructure; D003486:Cyanides / Q000494:pharmacology; D003576:Electron Transport Complex IV / Q000032:analysis*; D008928:Mitochondria / Q000187:drug effects / Q000201:enzymology*; D010477:Perfusion; D051381:Rats; D019265:Spectroscopy, Near-Infrared / Q000379:methods*; D013552:Swine
D003486:Cyanides; D003576:Electron Transport Complex IV