Related Experiment Videos
Double staining of proteoglycans and the HNK-1 carbohydrate epitope in pseudoexfoliation material
1Department of Ophthalmology, Faculty of Medicine, Kyushu University, Fukuoka, Japan. kubota@eye.med.kyushu-u.ac.jp
Purpose:
We combined the techniques of both immunogold and cupromeronic-blue staining with electron microscopy to investigate the possible relationship between proteoglycans and the HNK-1 carbohydrate epitope in pseudoexfoliation material.
Methods:
The anterior segment was obtained from an enucleated eye with pseudoexfoliation syndrome. After cupromeronic-blue staining, following the method of Scott, specimens were embedded in L. R. White resin. Ultrathin sections were incubated successively in primary monoclonal antibodies, and 10 nm gold-conjugated secondary antibody.
Results:
The present study showed simultaneous presence 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 pseudoexfoliation fibers, while cupromeronic blue-positive filaments bridged adjacent pseudoexfoliation fibers.
Conclusions:
Proteoglycans and 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 material.