Abstract
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 Cd 2+ 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 Ca 2+-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.
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