DOUBLE STAINING OF PROTEOGLYCANS AND THE HNK-1 CARBOHYDRATE EPITOPE IN PSEUDOEXFOLIATION MATERIAL

Purpose. We combined the techniques of both immunogold and cupromeroni c-blue straining with electron microscopy to investigate the possible relationship between proteoglycans and thee I-INK-I carbohydrate epito pe in pseudoexfoliation material. Methods. The anterior segment was ob tained from an enucleated eye with pseudoexfoliation syndrome. After c upromeronic-blue staining, following the method of Scott, specimens we re embedded in L. R. White resin. Ultrathin sections were incubated su ccessively in primary monoclonal antibodies, and 10 nm gold-conjugated secondary antibody. Results. The present study showed simultaneous pr esence of the HNK-1 epitope and proteoglycans associated with zonular fibrils and pseudoexfoliation material on the posterior surface of the iris. The HNK-1 epitope was immunolocalized to the periphery of the p seudoexfoliation fibers, while cupromeronic blue-positive filaments br idged adjacent pseudoexfoliation fibers. Conclusions. Proteoglycans an d the HNK-1 epitope might play a substantial role In the formation of pseudoexfoliation material via their adhesive function. Proteoglycans probably do not directly bear the HNK-1 epitope in pseudoexfoliation m aterial.