Opsin synthesis and mRNA levels in dystrophic retinas devoid of outer segments in retinal degeneration slow (rds) mice.
Opsin gene regulation, as a function of outer segment structure, was studied in normal and mutant retinal degeneration slow (rds) mice. We investigated the level of expression of the opsin gene in the rds mutant to determine if the reduced opsin content observed in this mutation (around 3% of normal) is a consequence of lowered expression of its gene. Normal BALB/c and rds mice were analyzed for levels of opsin mRNA and opsin content by Northern and immunoblot analysis, respectively. The rate of opsin synthesis in isolated retinas was measured by 35S-methionine incorporation in vitro, followed by analysis of the radiolabeled opsin by SDS-gel electrophoresis and autoradiography. Photoreceptor cell loss at various stages of degeneration was determined by quantitation of surviving photoreceptor nuclei. Opsin was localized in the mutant photoreceptors by immunoelectron microscopy of LR gold-embedded retinas using anti-opsin and antibody gold conjugates. The results indicate that 11- and 30-d-old mutant mice have considerable levels of opsin mRNA (60-70% of normal) and opsin synthetic rates (76-92% of normal), after the data from mutant mice are corrected for photoreceptor cell loss. We conclude, therefore, that the very low level of opsin observed in rds mice (approximately 3%) is not a result of greatly reduced expression of the opsin gene. Rather, continuous turnover of newly synthesized opsin as a result of its failure to become sequestered into an intact outer segment appears to account for the low levels of opsin in the rds mutant.
Agarwal
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Department of Pathology, University of Texas Health Science Center, San Antonio 78284.
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Papermaster
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D000818:Animals; D002467:Cell Nucleus / Q000473:pathology; D015342:DNA Probes; D005136:Eye Proteins / Q000096:biosynthesis* / Q000235:genetics; D015870:Gene Expression; D015151:Immunoblotting; D007150:Immunohistochemistry; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008817:Mice, Mutant Strains; D016253:Microscopy, Immunoelectron; D009154:Mutation; D009693:Nucleic Acid Hybridization; D010786:Photoreceptor Cells / Q000378:metabolism / Q000648:ultrastructure; D012333:RNA, Messenger / Q000378:metabolism*; D012162:Retinal Degeneration / Q000235:genetics* / Q000473:pathology; D012374:Rod Cell Outer Segment / Q000473:pathology*; D017299:Rod Opsins
D015342:DNA Probes; D005136:Eye Proteins; D012333:RNA, Messenger; D017299:Rod Opsins