Expression of opsin and IRBP genes in mutant RCS rats.
The retinal pigment epithelium of RCS rats bearing the autosomal recessive rdy mutation fails to ingest shed rod outer segment tips. Accumulation of disk debris in the subretinal space of the maturing mutant retina causes a secondary degeneration of photoreceptor cells. Two hypotheses have been offered as possible explanations of the death of photoreceptor cells in this disorder: (1) photoreceptors are starved for amino acids, retinal, oxygen, etc; and (2) that IRBP levels and synthesis may be decreased and interfere with retinal transport and this deficiency is lethal to these cells. To test these hypotheses, we have studied the effect of this mutation on the levels of expression of opsin and IRBP genes, and gene products and on rates of synthesis at various ages in dystrophic RCS p+ rats and compared the results to those obtained with normal Long Evans rats. The mutant rats and normal controls had comparable amounts of opsin and IRBP mRNA transcripts and rates of synthesis up to post-natal day 45 (P45) but opsin transcripts were barely detectable at P60 and thereafter. IRBP mRNA levels were also very low after P62 although somewhat higher than opsin mRNA. Opsin could be detected immunochemically, albeit at lower levels, at all the ages studied up to P310, but IRBP levels fell below detection after P45. We localized opsin and IRBP in the retina by post-embedding EM immunocytochemical procedures and found that opsin is present in the remnants of rod outer segment debris, even at P390, long after detectable opsin synthesis had ceased. These data suggest that expression of opsin and IRBP genes is not influenced by the shape and state of the outer segments, and that the rdy mutation does not influence the expression of the opsin and IRBP in these retinas until the photoreceptor cells are profoundly damaged. Thus, neither hypothesis about the causes of cell death in this disorder is supported.
Agarwal
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Department of Pathology, University of Texas Health Science Center, San Antonio 78284-7750.
Nir
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Papermaster
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D000375:Aging / Q000378:metabolism; D000818:Animals; D005136:Eye Proteins / Q000096:biosynthesis / Q000235:genetics*; D015151:Immunoblotting; D008854:Microscopy, Electron; D010786:Photoreceptor Cells / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D011922:Rats, Mutant Strains; D012160:Retina / Q000648:ultrastructure; D012177:Retinol-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D017299:Rod Opsins; D000099214:Retinol-Binding Proteins, Interstitial
D005136:Eye Proteins; D012333:RNA, Messenger; D012177:Retinol-Binding Proteins; D017299:Rod Opsins; D000099214:Retinol-Binding Proteins, Interstitial