Lectin-ferritin binding on dystrophic and normal retinal pigment epithelial membranes

Phagocytosis in the rat retina is a process which involves uptake of shed photoreceptor outer segments by the overlying retinal pigment epithelium (RPE). In rats with inherited retinal degeneration, there is a defect in phagocytosis. One aspect of this phagocytic defect may be an alteration in glycoconjugate-containing membrane components on RPE membranes which mediate this phagocytic interaction. Lectin binding sites have been studied in order to localize the distribution of glycoconjugates on pigment epithelial microvilli in normal and dystrophic retinas and to determine if there are differences in the dystrophic retinas which would provide a clue about the defect. The following ferritin-conjugated lectins were used in this study: Concanavalin A (Con A-fe) or lens culinaris haemagglutinin (LcH-fe) for mannosyl-containing glycoconjugates; and wheat germ agglutinin (WGA-fe) for n-acetylglucosamine and sialic acid-containing glycoconjugates. Control tissue was incubated in the lectin in the presence of its competitor sugar. The number of ferritin particles or lectin-ferritin binding sites per micrometre of microvillous membrane was quantified from electron micrographs using a computer and a digitizing tablet. The number of WGA-fe binding sites on normal RPE microvillous membranes (56.0/μm) was statistically equivalent to the dystrophic membranes (48.8/μm). The number of Con A-fe binding sites on normal (27.3/m) and dystrophic RPE (26.7/μm) was also the same. A dramatic difference in LcH-fe binding sites was demonstrated on normal (1.5/μm) as compared to dystrophic RPE (19.1/μm). Our results indicate that more mannosyl residues are accessible on dystrophic microvillous membranes and, based on what is currently known about LcH binding, that these residues belong to glycoconjugates having fucosyl-containing carbohydrate cores. The data also suggest that in normal animals without a phagocytic defect such fucosyl-containing glycoconjugates are not as accessible and may be masked by other sugar residues in the oligosaccharide chain.