This project is concerned with the thorough identification and characterization of transport ATPases in the corneal endothelium that may be directly or indirectly related to deturgescence. It is directed toward revealing some of the biochemical nature of the dehydration pumps (i.e., deturgescence) and factors that may control the pumps under normal, stressed and therapeutic conditions. Sodium-potassium stimulated ATPase and non-specific magnesium stimulated ATPase will be further characterized in the plasma membranes of bovine corneal endothelial cells. Anion-stimulated ATPase will be further characterized in the mitochrondria of bovine corneal endothelial cells. Agents known to stimulte or suspected to stimulate deturgescence will be used to test alterations in the activities of these enzymes. Agents used for pharmacological intervention of ocular function will be tested for their possible inhibitory effects on the activities of these enzymes. The activities of the ATPase found in the plasma membranes and mitochondria of corneal endothelial tissue cultures will be compared with the activities found in fresh tissue. Assays will be run by the measurement of inorganic phosphate released from the enzymatic hydrolysis of ATP and compared to tissue protein levels.

National Institute of Health (NIH)
National Eye Institute (NEI)
Research Project (R01)
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Visual Sciences A Study Section (VISA)
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University of Alabama Birmingham
Schools of Optometry/Opht Tech
United States
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Blake, D A; Whikehart, D R; Yu, H et al. (1996) Common cryopreservation media deplete corneal endothelial cell plasma membrane Na+,K+ ATPase activity. Curr Eye Res 15:263-71
Whikehart, D R; Angelos, P; Montgomery, B (1995) Effects of mannitol on cultured corneal endothelial cell Na,K-ATPase activity. Cornea 14:295-9
Whikehart, D R; Montgomery, B; Wells, J D et al. (1994) Low-molecular-weight peptides in corneal tissue culture media and in bovine aqueous fluid. Ophthalmic Res 26:17-22
Vogel, T; Blake, D A; Whikehart, D R et al. (1993) Specific simple sugars promote chemotaxis and chemokinesis of corneal endothelial cells. J Cell Physiol 157:359-66
Whikehart, D R; Montgomery, B; Angelos, P et al. (1993) Alteration of ATPase activity and duplex DNA in corneal cells grown in high glucose media. Cornea 12:295-8
Whikehart, D R; Montgomery, B; Sorna, D H (1992) The inhibition of Na, K-ATPase, and Mg-ATPase by timolol maleate in cultured non-pigmented epithelial cells of the ciliary body. J Ocul Pharmacol 8:107-14
Whikehart, D R; Montgomery, B; Sorna, D H et al. (1991) Beta-blocking agents inhibit Na+K+ATPase in cultured corneal endothelial and epithelial cells. J Ocul Pharmacol 7:195-200
Whikehart, D R; Edwards, W C; Pfister, R R (1991) Sorption of sodium hydroxide by type I collagen and bovine corneas. Cornea 10:54-8
Whikehart, D R; Montgomery, B; Wells, J D (1989) Age as a function of ATPase activity in cultured corneal endothelial cells. Invest Ophthalmol Vis Sci 30:335-8
Whikehart, D R; Montgomery, B; Hafer, L M (1987) Sodium and potassium saturation kinetics of Na+K+-ATPase in plasma membranes from corneal endothelium: fresh tissue vs. tissue culture. Curr Eye Res 6:709-17

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