Effect of “flow anoxia” and “non flow anoxia” on the NAD/NADH redox state of the intact brain cortex of the cat

Abstract

In the present study, we compared the nicotin-amide adenine dinucleotide (NAD) reducing potencies of “flow anoxia” and “non flow anoxia” in the cat brain cortex. In animals anaesthetized with alpha-D-glucochloralose “flow anoxia” and “non flow anoxia” were produced by ventilating for 2 and 25 min, respectively, with nitrogen gas. Following “non flow anoxia”, the brain cortices of dead animals were superfused with oxygen saturated artificial cerebrospinal fluid (mock CSF), and subsequently with CSF containing various concentrations (10⁻³–10⁻¹ M) of potassium cyanide. NADH (reduced NAD) fluorescence of the brain cortex was measured through a cranial window with microscope fluororeflectometer. Ventilating the animals for 2 and 25 min with nitrogen gas increased cortical NADH fluorescence (NAD reduction) by 43.5±2.8% and 135.3±6.1%, respectively. Oxygen saturated CSF superfusion of the ischemic brain cortex restored the cortical NAD/NADH redox state to the preanocic level (oxidation of NADH). 10⁻¹ M cyanide, applied after superfusion of the brain cortex with oxygen saturated CSF resulted in comparable NAD reduction to that produced by “non flow anoxia”. On the basis of these findings it is suggested that “non flow anoxia” leads to much greater cortical NAD reduction than “flow anoxia”, because oxygen tension in the cortex may not fall to zero mm Hg during nitrogen anoxia lasting for 2 min. Besides this, a more pronounced substrate mobilization and acidosis may also contribute to the greater NAD reducing potency of “non flow anoxia”. Finally, since 10–15 min after the death of the animal the cerebral carbohydrate reserves are completely exhausted, and in our experiments “non flow anoxia”, reoxygenation of the ischemic brain cortex and inhibition of the cortical mitochondrial electron transport by cyanide (10⁻¹ M) resulted in comparable redox state changes (as far as their magnitude is concerned), it is concluded that the recorded changes in NADH fluorescence were of mitochondrial origin.