The long-term objective of this proposal is to test the hypothesis that specific growth factors, and possibly other glycated polypeptides, which are present in the aqueous humor modulate the functional integrity of the aqueous outflow pathway in health and disease.
The specific aims are: (1) To quantify the amounts of two growth factors, basic fibroblast growth factor (b-FGF) and transferrin, that are present in normal primary aqueous humor of the human eye. (2) To determine the quantitative and qualitative differences in these growth factors, as well as in other glycated polypeptides, in secondary aqueous humor as compared to primary aqueous humor, by using the cat as an experimental model. (3) To analyze the types and amounts of growth factors in the aqueous humor obtained from patients with various diseases of the eye, including glaucoma. (4) To test in vitro whether specific molecules that are present in the aqueous humor stimulate the growth of cells of the trabecular meshwork and of the fibroblasts of Tenon's capsule. The health-relatedness of the project is: (1) to explore the feasibility of using growth factors for the control of age-related disorders of the anterior segment of the eye, especially primary open-angle glaucoma; and (2) to control the post-surgical over-growth and synthetic activity of trabecular meshwork cells and of Tenon's capsule fibroblasts, as well as neovascularization, that contribute to failure of filtering surgical procedures for glaucoma. The experimental design is to investigate in detail the qualities and quantity of B-FGF and transferrin, as well as other glycated polypeptides, that are present in primary and secondary aqueous humor and to evaluate their growth-promoting or -inhibiting properties. The methods to be used are: (1) dot/ slot binding and antibody-capture assay to quantitate b-FGF and transferrin; (2) a feline experimental model of primary and secondary aqueous humor for determination of the quantitative and quantitatives differences in these two growth factors; (3) SDS-PAGE and lectin binding on Western blots to evaluate the glycated polypeptides in primary and secondary aqueous humor from cat eyes, as well as in aqueous humor from patients with various diseases of the eye, including glaucoma; and (4) tissue culture, morphologic criteria, cell counting, rate of mitosis, and radioimmunoassay of newly synthesized polypeptides to investigate the effect of b-FGF and transferrin on primary cell cultures of trabecular meshwork cells and Tenon's capsule fibroblasts.
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