Mechanisms of the photo-damage of a fluorescent dye (methylene blue) and of the protective action of antioxidants and quenchers of singlet oxygen on the outer retinal rod segments (ORRS) and retinal function in situ and in vivo were studied. The methylene blue-induced formation of singlet oxygen in the ORRS resulted in accumulation of lipid peroxidation products, oligomerization of rhodopsin, and in a decrease in rhodopsin thermal resistance. Modification of the lipid and protein components of the visual cells by singlet oxygen inhibited the electrical activity of both isolated frog retina in situ and rabbit retina in vivo (waves a and b on the electroretinogram). The antioxidants (alpha-naphthol, alpha-tocopherol, 4-methyl-2,6-di-tert-butylphenol) and singlet oxygen quenchers, [1,4-diazabicyclo (2,2,2)octane] and alpha-tocopherol prevented the damaging effects of the fluorescent dye induced by singlet oxygen formation.
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