Bovine rod rhodopsin. 1. Bleaching by luminescence in vitro by recombination of radicals from polyunsaturated fatty acids.
Rod outer segments of photoreceptors are characterized by rhodopsin, a membrane protein surrounded by phospholipids containing a very high concentration of polyunsaturated fatty acids. These fatty acids can propagate free radicals, initiated by peroxidation, whose recombination is eventually associated with light emission as chemiluminescence. The results reported here indicate that this effect produces an isomerization of the retinal (bleaching effect) of the rhodopsin, similar to that induced by light in normal vision. In vitro experiments on detergent-suspended rod outer segments (RdOS) from bovine eyes, using an enzymatic source of radicals, xanthine/xanthine oxidase, were carried out. The results indicate that the proposed mechanism is likely, because they can show the bleaching of rhodopsin in RdOS, owing to its extraordinary sensitivity. Thus this mechanism is, also, a possible explanation for anomalous visual effects such as light flashes (phosphene-like) perceived by humans. The functionality of the rhodopsin in the RdOS was first tested by visible light. Rhodopsin reactivation after bleaching was obtained by adding cis-retinal to the suspension, demonstrating the reversibility of the bleaching process. A special experimental system was developed to observe the bleaching from luminescence by radical recombination, avoiding physical contact between the rod outer segment suspension and the radicals to prevent radical-induced damage and modifications of the delicate structure of the rod outer segment.
Department of Physics, University of Rome Tor Vergata, Rome, Italy.
L
L
Narici
M
M
Paci
V
V
Brunetti
A
A
Rinaldi
W G
WG
Sannita
A
A
De Martino
D000818:Animals; D002417:Cattle; D005231:Fatty Acids, Unsaturated / Q000737:chemistry / Q000378:metabolism*; D016166:Free Radical Scavengers / Q000494:pharmacology; D005609:Free Radicals / Q000737:chemistry / Q000378:metabolism*; D005978:Glutathione / Q000494:pharmacology; D007536:Isomerism; D015227:Lipid Peroxidation; D049449:Luminescence; D038761:Photobleaching; D012243:Rhodopsin / Q000737:chemistry / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000378:metabolism / Q000528:radiation effects*; D019820:Xanthine / Q000737:chemistry; D014969:Xanthine Oxidase / Q000737:chemistry
D005231:Fatty Acids, Unsaturated; D016166:Free Radical Scavengers; D005609:Free Radicals; D019820:Xanthine; D012243:Rhodopsin; D014969:Xanthine Oxidase; D005978:Glutathione