The aims of the project are to understand the mechanisms, cellular and molecular by means of which the opercular epithelium, corneal epithelium, ciliary epithelium and retinal pigment epithelium perform their functionsof ion and non-electrolyte transport and water. The consequences of the study will lead to further comprehension of membrane basic mechanisms that can lead to production of eventual therapeutic conduct. The regulatory process controlled by specific receptor proteins in the cells membranes and the channels affecting traffic across these membranes will provide a detailed account that will permit the formulation of eventual modes of control in the function of the corneal epithelium, the opercular epithelium, the production of aqueous humor and the control of the microenvironment of the retina. The methods to be used are of two natures: (1) electrophysiological and water flow methodology consisting in the use of transepithelial current and flux measurements, use of specific intracellular electrodes for C1, Na and Ca, together with the use of the new method of voltage patching. These measurements in different experimental conditions will provide the function of the apical and basolateral membrane of the epithelial cells with respect to ion movements and the effects of drugs and specific agents controlling channels and pumps. The water movements of isolated epithelia will be determined with a nanoliter injector method permitting electrical measurements simulataneously. The combination of these methods will provide a cellular picture for the epithelial functions and their modifications in altered states. (2) The molecular approach will consist of the use mainly of fluorescently labelled probes of receptors and channels. These biochemical or physicochemical approaches will use whole cells obtained from the epithelia of cell membrane fragments. Proteins will be purified, such as specific receptors and their properties and incorporation into cell membranes followed by fluorescence spectroscopy or microscopy. The characterization of these membrane proteins will require the preparation of specific antibodies, use of cultures and the utilization of monoclonally produced antibodies.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM025002-09
Application #
3272718
Study Section
Physiology Study Section (PHY)
Project Start
1978-04-01
Project End
1987-12-31
Budget Start
1986-07-01
Budget End
1987-12-31
Support Year
9
Fiscal Year
1986
Total Cost
Indirect Cost
Name
New York University
Department
Type
Schools of Medicine
DUNS #
004514360
City
New York
State
NY
Country
United States
Zip Code
10012
Zadunaisky, J A; Kinne-Saffran, E; Kinne, R (1989) A Na/H exchange mechanism in apical membrane vesicles of the retinal pigment epithelium. Invest Ophthalmol Vis Sci 30:2332-40
Flugel, C; Lutjen-Drecoll, E; Zadunaisk, J A et al. (1989) Regional differences in the morphology and enzyme distribution of the spiny dogfish (Squalus acanthias) ciliary epithelium. Exp Eye Res 49:1097-114
Jessen, F; Cherksey, B D; Zeuthen, T et al. (1989) Isolation and reconstitution of furosemide-binding proteins from Ehrlich ascites tumor cells. J Membr Biol 108:139-51
Wiederholt, M; Flugel, C; Lutjen-Drecoll, E et al. (1989) Mechanically stripped pigmented and non-pigmented epithelium of the shark ciliary body: morphology and transepithelial electrical properties. Exp Eye Res 49:1031-43
Scheide, J I; Zadunaisky, J A (1988) Effect of a chloride channel blocker on bullfrog corneal epithelium. Am J Physiol 254:C519-25
Charney, A N; Scheide, J I; Ingrassia, P M et al. (1988) Effect of pH on chloride absorption in the flounder intestine. Am J Physiol 255:G247-52
Zeuthen, T; Andersen, P M; Eskesen, K E et al. (1988) Characterization of a purified co-transporting protein. Comp Biochem Physiol A Comp Physiol 90:687-91
Cherksey, B D (1988) Functional reconstitution of an isolated sodium channel from bovine trachea. Comp Biochem Physiol A Comp Physiol 90:771-3
Scheide, J I; Zadunaisky, J A (1988) Effect of atriopeptin II on isolated opercular epithelium of Fundulus heteroclitus. Am J Physiol 254:R27-32
Zadunaisky, J A; Curci, S; Schettino, T et al. (1988) Intracellular voltage recordings in the opercular epithelium of Fundus heteroclitus. J Exp Zool 247:126-30

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