Freeze-fracture label of functional and dysfunctional human corneal endothelium

Previous freeze-fracture results from our laboratory have shown a reduction in a population of intramembrane particles in the lateral endothelial membranes from dysfunctional human corneas. The size range of these intramembrane particles corresponds to that which has previously been reported for the glycoprotein enzyme Na, K-ATPase in enzyme enriched freeze-fractured membranes. In order to investigate glycoconjugate changes potentially related to the particle reduction, wheat germ agglutinin (WGA), which has been shown to bind to the sugar residues in the ATPase subunit, was used to label three types of corneas with dysfunctional endothelial cells (Fuchs' endothelial dystrophy, aphakic and pseudophakic bullous keratopathy) and two types of corneas (eye bank and keratoconus) with functional endothelium using the technique of thin section freeze-fracture label. Apical WGA labelling on all types of dysfunctional cells was shown to be drastically reduced in comparison to both types of functional corneal endothelial cells. Lateral membranes of dysfunctional cells, exposed by freeze-fracturing, also showed a great reduction in WGA labelling as compared to the fractured lateral membranes of functional cells. The differences observed in lectin labelling of lateral membranes may be related in part to the decreased intramembrane particle density observed in dysfunctional human corneal endothelial cells.