A biomolecular approach to the study of the expression of specific genes in the retina

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Abstract
The central nervous system contains a number of populations of neurons that are morphologically and functionally distinct. To study the genes responsible for the development and maintenance of proteins with unique structural and biochemical properties, polyspecific antisera were produced against normal mouse retinal proteins and used in combination with the rd (retinal degeneration) neurological mutant of the mouse to immunologically identify specific retinal proteins. Western transfer analysis of the proteins present in normal and in mutant retinas identified three classes of neural proteins: (1) those found only in normal retina; (2) those found in normal and in photoreceptorless retinas but not in other tissues; and (3) those found in both normal and mutant retinas, as well as in brain, but not elsewhere. Some of these class 1 proteins were shown to be present in the retinas of other species, including humans, suggesting their importance in the process of vision. The poly A + RNA was isolated from the retinas of normal mice and used to generate a cDNA expression library in lambda gt-11. This library was screened with polyspecific antisera absorbed with the proteins present in mutant retina, and a number of immunologically positive plaques were cloned. Four of these were shown to code for rhodopsin, the major visual protein in mammalian retinas. The approach described is applicable to other systems in order to generate specific immunological and recombinant DNA probes for examining the expression of specific genes during development and differentiation.