Cytochrome C oxidase expression in retinal rod outer segment disks

Abstract

Abstract Purpose The rod outer segment (ROS) plays a pivotal role in the phototransduction cascade, a process which requires a high level of energy consumption. In rods ATP production is confined to the mitochondria in the inner part, and the distances between sites of energy production and expenditure do not seem adequate to supply phototransduction. The goal of this study was to test the hypothesis that Cytochrome c oxydase (Cox) is located and catalytically active in ROS disk membranes. Methods Osmotically intact disks were isolated from 20 bovine retinal ROS obtained by sucrose gradient centrifugation in the presence of protease inhibitor cocktail and Ampicillin under dim red light at 4°C. Cox expression was studied by confocal laser scanning microscopy and semiquantitative Western‐immunoblotting, with a specific antibody. Cox activity in disks was investigated spectrophotometrically. Results Results from immunohystochemical confocal laser scanning microscopy, Western blot, and biochemical experiments suggest that Cox is uniformly distributed onto disks and therein catalytically active to an extent comparable to that of isolated mitochondria. Conclusion The expression and activity of Cox in bovine ROS disks is suggestive of an aerobic metabolism in ROS. This study opens new scenarios on the energy production in rods and on the possible role of its deficiency in some important retinal diseases which have not been clarified yet.