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Am J Physiol Cell Physiol 276: C659-C666, 1999;
0363-6143/99 $5.00
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Vol. 276, Issue 3, C659-C666, March 1999

A3 adenosine receptors regulate Clminus channels of nonpigmented ciliary epithelial cells

Claire H. Mitchell1, Kim Peterson-Yantorno1, David A. Carré1, Alice M. McGlinn2, Miguel Coca-Prados3, Richard A. Stone2, and Mortimer M. Civan1,4

Departments of 1 Physiology, 2 Ophthalmology, and 4 Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-6085; and 3 Department of Ophthalmology and Visual Science, Yale University School of Medicine, New Haven, Connecticut 06510

Adenosine stimulates Cl- channels of the nonpigmented (NPE) cells of the ciliary epithelium. We sought to identify the specific adenosine receptors mediating this action. Cl- channel activity in immortalized human (HCE) NPE cells was determined by monitoring cell volume in isotonic suspensions with the cationic ionophore gramicidin present. The A3-selective agonist N6-(3-iodobenzyl)-adenosine-5'-N-methyluronamide (IB-MECA) triggered shrinkage (apparent Kd = 55 ± 10 nM). A3-selective antagonists blocked IB-MECA-triggered shrinkage, and A3-antagonists (MRS-1097, MRS-1191, and MRS-1523) also abolished shrinkage produced by 10 µM adenosine when all four known receptor subtypes are occupied. The A1-selective agonist N6-cyclopentyladenosine exerted a small effect at 100 nM but not at higher or lower concentrations. The A2A agonist CGS-21680 triggered shrinkage only at high concentration (3 µM), an effect blocked by MRS-1191. IB-MECA increased intracellular Ca2+ in HCE cells and also stimulated short-circuit current across rabbit ciliary epithelium. A3 message was detected in both HCE cells and rabbit ciliary processes using RT-PCR. We conclude that human HCE cells and rabbit ciliary processes possess A3 receptors and that adenosine can activate Cl- channels in NPE cells by stimulating these A3 receptors.

aqueous humor secretion; chloride channels; N6-(3-iodobenzyl)-adenosine-5'-N-methyluronamide; MRS-1097; MRS-1191; MRS-1523


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