Opsin distribution and synthesis in degenerating photoreceptors of rd mutant mice

Abstract

The distribution of opsin and the capacity of photoreceptors to synthesize opsin was studied in retinas of mice bearing the rd (retinal degeneration) mutation and compared to control normal mice. Opsin was localized by means of pre-embedding and post-embedding immunocytochemistry and electron microscopy. Cones were identified with anti-cone antibodies and peanut agglutinin lectin which labels cone matrix sheaths. Opsin synthesis was measured by incorporation of [ 35S]methionine into opsin which was detected by immunoblots. Immunocytochemistry revealed that degeneration of rod outer segments was accompanied by accumulation of opsin in the plasma membrane enveloping the inner segment, nuclei and synaptic terminals. Rod photoreceptors degenerated faster than cones. By post-natal day 19 (P19), 87% of the remaining inner segments were cones. Opsin synthesis in rd mice could no longer be measured after P15. However, opsin molecules could be detected both by immunocytochemistry and immunoblotting up to P30. Between P20 and P30 all detectable opsin was localized in the plasma membrane which envelopes nuclei and synaptic terminals. Unlabeled surviving nuclei after P30 are probably cones. The opsin which is detected in rod inner segment plasma membranes might be derived, by back diffusion, from degenerating outer segments and represent molecules synthesized at an earlier time. Alternatively opsin accumulation might be a result of low levels of opsin synthesis, undetectable by [ 35S]methionine incorporation, which continues in the absence of outer segments.