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Am J Physiol Cell Physiol 275: C1022-C1030, 1998;
0363-6143/98 $5.00
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Vol. 275, Issue 4, C1022-C1030, October 1998

Energy-dependent redox state of heme a + a3 and copper of cytochrome oxidase in perfused rat brain in situ

Akira Matsunaga1,2, Yasutomo Nomura1, Satoshi Kuroda1, Mamoru Tamura1, Jun Nishihira3, and Nozomu Yoshimura2

1 Biophysics Group, Research Institute for Electronic Science, Hokkaido University, and 3 Central Research Institute, Hokkaido University School of Medicine, Sapporo 060; and 2 Department of Anesthesiology and Critical Care Medicine, Kagoshima University School of Medicine, Kagoshima 890, Japan

Using the blood-free perfused rat brain, we examined the redox behavior of cytochrome oxidase of two chromophores, heme a + a3 and copper. When perfusate inflow was stopped to induce global ischemia, the reduction of heme a + a3 was triphasic, with a rapid phase, a slow phase, and a second rapid phase. In contrast, the reduction of copper was monophasic after the rapid phase of heme a + a3. The triphasic reduction of heme a + a3 was diminished by energy-depleting treatments, such as addition of an uncoupler. The time course of the reduction of copper was not affected by the energy depletion. During global ischemia the decrease in creatine phosphate nearly paralleled the reduction of heme a + a3, whereas ATP remained at the control level until ~60% of heme a + a3 was reduced in the rapid phase. In the slow phase, ATP started to decrease with the reduction of copper. The redox behavior of copper was similar to the slow phase of the reduction of heme a + a3 because of the higher oxygen affinity of copper than of heme a + a3. Therefore, the rapid phase of the reduction of heme a + a3 can be used as an alarm before a decrease in ATP, whereas the reduction of copper indicates a decrease in ATP under severe hypoxia. Thus the copper signal in noninvasive near-infrared spectroscopy is a useful parameter for the clinical setting.

oxygen supply; mitochondria; uncoupled state; hypoglycemia; near-infrared spectroscopy


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