The PI proposes that a novel paracrine system based on purinergic signaling allows for intraepithelial communication between the non-pigmented (NPE) and pigmented (PE) ciliary epithelial cell layers. From the work of others, purinergic mechanisms are known to alter aqueous humor inflow and intraocular pressure. He specifically proposes that: (1) ATP (and possibly other nucleotides and adenosine) are released directly by NPE and PE cells; (2) Membrane ecto-enzymes hydrolyse ATP to AMP and extracellular adenosine; (3) Adenosine acts on NPE cells to stimulate aqueous humor formation by activating Cl- channels; (4) Extracellular ATP stimulates aqueous humor reabsorption by activating PE anion channels; and (5) Net aqueous humor formation is regulated by coordinating these opposing purinergic effects (and other less well defined transport effects) on unidirectional secretion and reabsorption. This coordination may reflect synergistic effects of purines and an unidentified intracellular regulator, whose action is mimicked by tamoxifen. The PI will address the following specific aims: (i) What are the characteristic biophysical and pharmacologic properties of the channels targeted by purines, especially the NPE Cl- channels and PE anion channels? (ii) What are the mode and trigger for purine release by the NPE and PE ciliary epithelial cells? (iii) What are the receptors and signaling pathways underlying purinergic stimulation of transport by the NPE and PE cell? (iv) How are these purinergic effects coordinated to modulate net aqueous humor secretion? The proposed interdisciplinary research will combine patch-clamp and volumetric physiologic methods with molecular biological and structural strategies, correlating measurements from isolated-cell, broken-cell and cell-couplet preparations with those from whole tissues. The novel hypothesis (that aqueous humor formation is directly modulated by an interlayer paracrine release of purinergic messengers) can provide the basis for understanding the purinergic effects on aqueous humor dynamics.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
1R01EY012213-01
Application #
2669592
Study Section
Visual Sciences A Study Section (VISA)
Project Start
1998-08-01
Project End
2001-07-31
Budget Start
1998-08-01
Budget End
1999-07-31
Support Year
1
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of Pennsylvania
Department
Physiology
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Mitchell, C H (2001) Release of ATP by a human retinal pigment epithelial cell line: potential for autocrine stimulation through subretinal space. J Physiol 534:193-202
Avila, M Y; Stone, R A; Civan, M M (2001) A(1)-, A(2A)- and A(3)-subtype adenosine receptors modulate intraocular pressure in the mouse. Br J Pharmacol 134:241-5
Avila, M Y; Carre, D A; Stone, R A et al. (2001) Reliable measurement of mouse intraocular pressure by a servo-null micropipette system. Invest Ophthalmol Vis Sci 42:1841-6
Fleischhauer, J C; Mitchell, C H; Peterson-Yantorno, K et al. (2001) PGE(2), Ca(2+), and cAMP mediate ATP activation of Cl(-) channels in pigmented ciliary epithelial cells. Am J Physiol Cell Physiol 281:C1614-23
Mitchell, C H; Peterson-Yantorno, K; Coca-Prados, M et al. (2000) Tamoxifen and ATP synergistically activate Cl- release by cultured bovine pigmented ciliary epithelial cells. J Physiol 525 Pt 1:183-93
Counillon, L; Touret, N; Bidet, M et al. (2000) Na+/H+ and CI-/HCO3-antiporters of bovine pigmented ciliary epithelial cells. Pflugers Arch 440:667-78