An assessment of rat photoreceptor sensitivity to mitochondrial blockade

To report results of functional, biochemical and structural studies of photoreceptor mitochondria in isolated rat retinas under conditions of mitochondrial inhibition.

Dark-adapted rat retinas were incubated in a modified Ringer's bicarbonate medium under aerobic and anaerobic conditions. Several different procedures were used to inhibit mitochondrial function; N2, 0.01 mM antimycin A, and 1 and 10 mM potassium cyanide (KCN). Measurements were made of lactic acid production, retinal adenosine triphosphate (ATP) content, and receptor potentials. Morphology of the inner segment mitochondria was examined by electron microscopy.

In the presence of N2, 0.01 mM antimycin, or 1 mM KCN, lactic acid production was linear throughout the 60- minute period; and the rate was similar for each condition. Retinal ATP content and the amplitude of the receptor potential were also maintained at high levels after short-term incubations with either N2, antimycin A, or 1 mM KCN. In contrast, use of 10 mM KCN produced an entirely different set of results. These effects were studied both at the alkaline pH (8.9) found when this concentration of KCN was simply added to bicarbonate-buffered media and at the normal pH (after readjustment) of 7.4. With 10 mM KCN (pH 8.9), retinal lactate production was severely depressed, retinal ATP content was nearly depleted within 5 to 10 minutes, and the amplitude of the receptor potential rapidly declined to a low level. The deleterious effects of 10 mM KCN on these parameters were lessened to varying degrees when pH was readjusted to 7.4. Electron microscopic observations of rat rod inner segments indicated generally excellent survival of these organelles after incubation with either N2, antimycin A, or 1 mM KCN in comparison with their appearance under oxygenated conditions. However, the inner segments were significantly disrupted after incubation of retinas with 10 mM KCN.

Findings suggest that the loss of the receptor potential and depletion of ATP observed with minutes after exposing isolated rat retinas to media containing 10 mM KCN results from the inhibition of both respiration and glycolysis by this high concentration of KCN. In contrast, when conditions are chosen so that only respiration is impaired (as with N2, antimycin A, or 1 mM KCN) photoreceptor cells are resistant to short-term episodes of mitochondrial inhibition, principally because the upregulation of glycolysis generates sufficient ATP to compensate reasonably well for the loss in mitochondrially produced ATP.