PMID: 7512946Apr 1, 1994Paper

Molecular and biochemical analyses of iodopsin in rd chick retina

Investigative Ophthalmology & Visual Science
Susan L Semple-Rowland, D A Green

Abstract

The results of previous immunocytochemical and electrophysiological studies of retinas of rd (retinal degeneration) chicks suggest that the iodopsin cone visual pigment may be defective in this mutant. The goal of this study was to determine if the primary structure and synthesis of this protein is normal in this animal model of inherited retinal degeneration. Northern cDNA sequence and western analyses were used to study rd/rd iodopsin. cDNAs encoding rd/rd iodopsin were obtained by screening an rd/rd cDNA retinal expression library and by reverse transcription PCR. Western blots were probed with either R4 or COS-1, two different monoclonal antibodies that have been shown to specifically recognize chicken iodopsin. Hybridization of the +/+, +/rd, and rd/rd poly(A)+ RNA with an iodopsin cDNA probe revealed the presence of a single 1.5 kb band in each of the samples, all of which were labeled with equal intensity. No significant differences were found between the published nucleic acid sequences for normal chicken iodopsin cDNA and that determined for rd/rd iodopsin cDNA. Antibody-dependent differences in the staining intensity of the 34 kDa band containing iodopsin were observed on western blots of +/+, +/rd, and rd/rd retinal ...Continue Reading

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