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(4)

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Abstract #15310 Published in IGR 8-4

Identification and functional characterization of ClC-2 chloride channels in trabecular meshwork cells

Comes N; Abad E; Morales M; Borras T; Gual A; Gasull X
Experimental Eye Research 2006; 83: 877-889

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In the eye, trabecular meshwork (TM) cell volume may be an important determinant of aqueous humor outflow. Among their functions, ClC-2 chloride channels are thought to be involved in regulation of cellular volume and intracellular [Cl(-)]. We characterized the properties and modulation of an inwardly rectifying chloride current activated in these cells. Patch-clamp recordings revealed inwardly rectifying chloride currents activated by membrane hyperpolarization in primary cultures of both bovine (BTM) and human (HTM) TM cells. Electrophysiological properties and anion permeability sequence (Cl^- > Br^- > I^- > F^- ) were in agreement with previous data for ClC-2 in other cells. The currents were blocked by Cd²⁺ and enhanced by extracellular acidification, 8Br-cAMP and cell swelling, while extracellular alkalinization decreased them. RT-PCR experiments using total RNA revealed the molecular expression of ClC-2 channels. Previously we reported the involvement of swelling-activated chloride channels (Cl_swell ) and Ca²⁺ -activated K⁺ channels (BK_Ca ) in cell volume and outflow facility regulation. However, in the present analysis, cell volume experiments in calcein-loaded cells and outflow facility studies performed in bovine anterior segments revealed that ClC-2 channels do not make a significant contribution to the recovery of cellular volume or to the regulation of the outflow facility. Nevertheless, ClC-2 modulation by different stimuli may contribute to intracellular [Cl^- ] regulation and other cellular functions yet to be determined in the TM.

Dr. N. Comes, Laboratory of Neurophysiology, Department of Physiological Sciences I-Institute of Biomedical Investigations August Pi i Sunyer, IDIBAPS, School of Medicine, University of Barcelona, Casanova 143, E-08036 Barcelona, Spain

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Classification:

2.5.1 Trabecular meshwork (Part of: 2 Anatomical structures in glaucoma > 2.5 Meshwork)
3.5 Molecular biology incl. SiRNA (Part of: 3 Laboratory methods)
3.6 Cellular biology (Part of: 3 Laboratory methods)

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