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(3)

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Abstract #3401 Published in IGR 4-2

Human trabecular meshwork cell volume regulation

Mitchell CH; Fleischhauer JC; Stamer WD; Peterson YK; Civan MM
American Journal of Physiology and Cell Physiology 2002; 283: C315-326

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The volume of certain subpopulations of trabecular meshwork (TM) cells may modify outflow resistance of aqueous humor, thereby altering intraocular pressure. This study examines the contribution that Na⁺/H⁺, Cl^-/HCO exchange, and K⁺-Cl^- efflux mechanisms have on the volume of TM cells. Volume, Cl- currents, and intracellular Ca²⁺ activity of cultured human TM cells were studied with calcein fluorescence, whole cell patch clamping, and fura 2 fluorescence, respectively. At physiological bicarbonate concentration, the selective Na⁺/H⁺ antiport inhibitor dimethylamiloride reduced isotonic cell volume. Hypotonicity triggered a regulatory volume decrease (RVD), which could be inhibited by the Cl^- channel blocker 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB), the K⁺ channel blockers Ba²⁺ and tetraethylammonium, and the K⁺-Cl^- symport blocker [(dihydroindenyl)oxy]alkanoic acid. The fluid uptake mechanism in isotonic conditions was dependent on bicarbonate; at physiological levels, the Na⁺/H⁺ exchange inhibitor dimethylamiloride reduced cell volume, whereas at low levels the Na⁺-K⁺-2Cl^- symport inhibitor bumetanide had the predominant effect. Patch-clamp measurements showed that hypotonicity activated an outwardly rectifying, NPPB-sensitive Cl^- channel displaying the permeability ranking Cl^- > methylsulfonate > aspartate. 2,3-Butanedione 2-monoxime antagonized actomyosin activity and both increased baseline [Ca²⁺] and abolished swelling-activated increase in [Ca²⁺], but it did not affect RVD. Results indicate that human TM cells display a Ca²⁺-independent RVD and that volume is regulated by swelling-activated K⁺ and Cl^- channels, Na⁺/H⁺ antiports, and possibly K⁺-Cl^- symports in addition to Na⁺-K⁺-2Cl^- symports.

Dr. C.H. Mitchell, Department of Physiology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-6085, USA

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Classification:

2.5 Meshwork (Part of: 2 Anatomical structures in glaucoma)
4 Tissue culture of ocular cells

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