Na+/H+ and Cl–/HCO3 –-antiporters of bovine pigmented ciliary epithelial cells

• Published in: Pflügers Archiv Vol. 440; no. 5; pp. 667 - 678
• Main Authors: Counillon, L., Touret, Nicolas, Bidet, Michel, Peterson-Yantorno, K., Coca-Prados, M., Stuart-Tilley, Alan, Wilhelm, Sabine, Alper, S.L., Civan, M.M.
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer Nature B.V 01.09.2000
• DOI: 10.1007/s004240000302
• ISSN: 0031-6768

Medical therapy of glaucoma commonly aims at slowing aqueous humor formation by the ocular ciliary epithelial bilayer, but underlying mechanisms are poorly understood. The first step in secretion is NaCl uptake from the stroma into the pigmented ciliary epithelial (PE) cell layer by electroneutral transporters. After crossing gap junctions into the nonpigmented ciliary epithelial (NPE) cell layer, solute is released into the aqueous humor. Published data have indicated that both paired Na^sup +^/H^sup +^ and Cl^sup -^/HCO^sub 3^^sup -^ antiporters and the Na^sup +^-K^sup +^-2Cl^sup -^ symporter are involved in net uptake. The molecular identities of the paired antiporters have not been elucidated. We have studied continuously cultured bovine PE cells. Acid-activated ^sup 22^Na^sup +^ uptake was inhibited by cariporide, EIPA (ethyl-isopropyl-amiloride) and amiloride, at concentrations characteristic of the NHE-1 isoform. Videomicroscopy of BCECF-loaded PE cells verified the presence of an EIPA-inhibitable Na^sup +^/H^sup +^ antiporter. Removing external Cl^sup -^ also triggered an alkalinization, which was Na^sup +^-independent and could be inhibited by 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS). Application of hypotonicity followed by return to isotonicity triggered a regulatory volume increase, which was pharmacologically similar to the uptake mechanisms described for intact rabbit ciliary epithelium. Reverse transcriptase polymerase chain reaction (RT-PCR) amplification of RNA from the human ciliary body detected expression of the AE2 Cl^sup -^/HCO^sub 3^^sup -^ exchanger, but not of AE1, cAE3 or bAE3. Immunostaining of bovine PE cells also revealed the presence of AE2 epitope. We conclude that paired NHE-1 Na^sup +^/H^sup +^ and AE2 Cl^sup -^/HCO^sub 3^^sup -^ antiporters are important components in the initial step in aqueous humor formation.[PUBLICATION ABSTRACT]