Immunohistochemical localization of cyclic GMP in goldfish retina.

Abstract

The immunocytochemical procedure for cyclic GMP (cGMP) localization has been modified by the development of: 1) an anti-cGMP antibody depleted control serum and 2) a tissue preadsorption procedure that greatly reduces nonspecific fluorescence. cGMP remaining associated with tissue sections following prolonged exposure to aqueous buffer was determined, by extraction and measurement by radioimmunoassay, to be approximately 38% of the initial cGMP content. Application of this improved information about histological distribution ofcGMP is already known from previous microdissection studies (9,10). immunohistochemical procedure to goldfish retinas reveals the highest concentrations of cGMP to be associated with the cone inner segments and nuclei of the photoreceptors. This retinal distribution of cGMP is in good agreement with that independently determined by microdissection studies. The validity of this procedure is therefore strengthened and cautions to its application and interpretation are outlined.

Morphological and physiological characteristics of dermal photoreceptors in <i>Lymnaea stagnalis </i>

“…We observed that the area at the outer segment of rods was immunopositive to rhodopsin antibody; the outer segment of photoreceptor layer, inner plexiform layer, ganglion cell layer was immunopositive to cGMP antibody; and the inner- and outer-segment of the photoreceptor layer was immunopositive to β-arrestin antibody (data not shown). These findings were consistent with previous observations 27 – 31 .…”

Immunocytochemistry of cGMP in the Cerebellum of the Immature, Adult, and Aged Rat: the Involvement of Nitric Oxide. A Micropharmacological Study

“…However, there are several problems associated with this method, the most important being the washing away of the cyclic nucleotides during the experimental procedure (Cumming et al, 1980;Ortez et al, 1980;Rall and Lehne, 1982). Using antibodies against the soluble guanylate cyclase the enzyme was shown to be associated with all cell types in the cerebellum (Ariano et al, 1982;Zwiller et al, 1981).…”

“…In the cerebellum cGMP was found associated with glial processes (Chan-Palay and Palay, 1979;Cumming et al, 1977Cumming et al, , 1979 by using the cGMP antibody as developed by Steiner et al (1972), in the immunocytochemical approach as introduced by Wedner et al (1972). However, there are several problems associated with this method, the most important being the washing away of the cyclic nucleotides during the experimental procedure (Cumming et al, 1980;Ortez et al, 1980;Rall and Lehne, 1982). Using antibodies against the soluble guanylate cyclase the enzyme was shown to be associated with all cell types in the cerebellum (Ariano et al, 1982;Zwiller et al, 1981).…”

Cytochemical demonstration of guanylate cyclase activity in retinal photoreceptors with special reference to changes under light and dark adaptation.

“…In this respect, information on histological distribution of cyclic GMP and measurement of cyclic GMP directly in photoreceptors in the retina after variation of the environment would be useful. Previous studies have been carried out concerning layer-by-layer analysis of quick-frozen eyes (2,19,36) or immunohistochemical localization of cyclic GMP (37). The ultrastructural localization of the enzymes required for cyclic nucleotide synthesis and degradation has been a matter of continuing interest.…”