Based on our previous investigations, the proposed continuation study of trabecular endothelium of normal and glaucomatous human eyes in a defined tissue culture environment seeks direct information concerning the morphology, pathophysiology, phagocytic activity, contractile protein content, and drug response of the cells. Throughout the investigation, comparable studies will be carried out on control cultures of corneal endothelium, keratocytes, and scleral fibroblasts. For tissue culture, human and monkey eyes will be used as soon as possible after enucleation. The morphology and behavior of the cultured trabecular cells will be studied in the living state by phase-contrast microscopy and time-lapse cinemicrography, and in fixed specimens by light and electron microscopy. The phagocytic activity of the cultured cells will be assessed by their uptake of latex spheres, carmine, and human erythrocytes introduced into the culture medium; the contractile proteins will be analyzed by electron microscopy, by immunofluorescence techniques, and by a radioimmunoassay, and quantification will be carried out by SDS electrophoresis and by a microassay method. We will use these parameters to determine the response of the cells to adrenergic agents and pilocarpine. Similar experiments will be carried out on the trabecular cells harvested from trabeculectomy specimens made available from glaucoma patients, undergoing elective surgery. Our overall objective in this project is to investigate systematically the morphology, and activity of the trabecular endothelium, and the manner in which it is able to regulate, facilitate, or obstruct aqueous outflow, with the prospect of ultimately gaining control of these interactions for therapy, or even prophylaxis, of glaucoma.
Showing the most recent 10 out of 31 publications
