Evidence for reduced binding of cyclic GMP to cyclic GMP phosphodiesterase in photoreceptors of mice heterozygous for the rd gene.

The binding of radiolabelled cGMP to rod outer segment proteins has been investigated in mice, heterozygous for the recessive rd gene that leads to rod dysplasia. Two binding sites were detected, by Scatchard analysis, in a crude cGMP phosphodiesterase fraction, extracted with an EDTA wash from outer segments. Affinities were normal but the capacity of both was reduced 25-35%. Photoaffinity labelling with 3H-cGMP, followed by SDS PAGE and fluorography, suggested that cGMP PDE was the principal binding component in the extracts. If the finding reflects cGMP binding in situ, it might explain the 30-40% lower than normal level of cGMP found in the +/rd retina. Visual pigment has been regenerated in isolated normal and heterozygotic retinas by the application of active isomers of cis-retinal, and the time course of cGMP recovery to 'dark-adapted' levels monitored. The increase in the concentration of cGMP was significantly delayed as compared to that of rhodopsin. No differences in time course or kinetics of recovery were discerned between the two genotypes.