Examination of sialic acid binding on dystrophic and normal retinal pigment epithelium.
In order to study differences in cell-surface sugars which may be involved in the phagocytic defect in Royal College of Surgeons (RCS) retinas, we have examined the presence or absence of lectin binding to carbohydrates on retinal pigment epithelial (RPE) plasma membranes of dystrophic (RCS-p+) and normal (Long-Evans) rats. A lectin which binds to both sialic acid and N-acetylglucosamine sugar residues, wheat germ agglutinin-ferritin (WGA-fe), was used. The specificity of WGA-fe binding to each sugar was studied by either pre-treating the tissue with neuraminidase enzyme which removes sialic acid residues, or by incubating the WGA-fe lectin with one of the haptens, N-acetylglucosamine. In non-enzyme-treated tissue, RPE cell-surface membranes from RCS retinas were densely labeled with WGA-fe as compared with the labeling on normal RPE, which appeared less dense and patchy in distribution. Wheat germ agglutinin-ferritin labeling in the presence of N-acetylglucosamine was blocked on both RCS and normal RPE surface membranes. After pre-treatment with neuraminidase, WGA-fe labeling on dystrophic RPE membranes was similar to non-enzyme-treated tissue but was enhanced on normal RPE. Labeling was blocked when N-acetylglucosamine was present with the lectin after enzyme pre-treatment. Other lectins which specifically bind to sialic acid, Limulus polyphemus agglutinin-ferritin (LPA-fe) and Limax flavus agglutinin (LFA) demonstrated sparse or no labeling on both RCS and normal RPE membranes. Our data suggests that N-acetylglucosamine residues predominate on RCS and normal RPE cell-surface membranes and that sialic acid binding sites are either not accessible to the lectins or may not be present.
D000117:Acetylglucosamine / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D005293:Ferritins / Q000378:metabolism; D009439:Neuraminidase / Q000378:metabolism; D010857:Pigment Epithelium of Eye / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012162:Retinal Degeneration / Q000378:metabolism*; D012794:Sialic Acids / Q000378:metabolism*; D014909:Wheat Germ Agglutinins / Q000378:metabolism
D012794:Sialic Acids; D014909:Wheat Germ Agglutinins; C053165:wheat germ agglutinin-ferritin; D005293:Ferritins; D009439:Neuraminidase; D000117:Acetylglucosamine