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Am J Physiol Cell Physiol 279: C440-C451, 2000;
0363-6143/00 $5.00
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Vol. 279, Issue 2, C440-C451, August 2000

Similarity of A3-adenosine and swelling-activated Clminus channels in nonpigmented ciliary epithelial cells

David A. Carré1, Claire H. Mitchell1, Kim Peterson-Yantorno1, Miguel Coca-Prados2, and Mortimer M. Civan1,3

Departments of 1 Physiology and 3 Medicine, School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104; and 2 Department of Ophthalmology and Visual Science, Yale University School of Medicine, New Haven, Connecticut 06510

Chloride release from nonpigmented ciliary epithelial (NPE) cells is a final step in forming aqueous humor, and adenosine stimulates Cl- transport by these cells. Whole cell patch clamping of cultured human NPE cells indicated that the A3-selective agonist 1-deoxy-1-(6-[([3-iodophenyl]methyl)amino]-9H-purin-9-yl)-N-methyl-beta -D-ribofuranuronamide (IB-MECA) stimulated currents (IIB-MECA) by ~90% at +80 mV. Partial replacement of external Cl- with aspartate reduced outward currents and shifted the reversal potential (Vrev) from -23 ± 2 mV to -0.0 ± 0.7 mV. Nitrate substitution had little effect. Perfusion with the Cl- channel blockers 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) and niflumic acid inhibited the currents. Partial Cl- replacement with aspartate and NO3-, and perfusion with NPPB, had similar effects on the swelling-activated whole cell currents (ISwell). Partial cyclamate substitution for external Cl- inhibited inward and outward currents of both IIB-MECA and ISwell. Both sets of currents also showed outward rectification and inactivation at large depolarizing potentials. The results are consistent with the concept that A3-subtype adenosine agonists and swelling activate a common population of Cl- channels.

aqueous humor secretion; anion selectivity; cyclamate; 1-deoxy-1-(6-[([3-iodophenyl]methyl)amino]-9H-purin-9-yl)-N-methyl-beta -D-ribofuranuronamide; MRS-1523


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