Abstract
Calcium-activated chloride currents were studied by the patch-clamp technique in vascular smooth muscle cells (VSMC) isolated from human mesenteric arteries. Bath application of 20 mM caffeine caused the cell membrane to depolarize by a calcium-activated inward current that peaked to -654 +/- 230 pA (holding potential -50 mV). Cell-attached, at the same time inwardly directed single-channel currents were detected with an amplitude of -0.22 pA. In open-cell-attached patches channel activity was triggered by elevating [Ca2+]i to 10 microM. At -60 mV the mean amplitude of the current was -0.24 pA and the mean open time of the channels was 28 ms. Plotting the amplitude of the current versus the test potential yielded a single-channel conductance of 2.8 +/- 0.5 pS. The currents disappeared when [Cl-] was reduced from 150 mM to 5 mM at the cytosolic side of the inside-out patch at a holding potential of -60 mV (calculated reversal potential -58 mV) suggesting that the calcium-activated current was a chloride current. This suggests that, in human mesenteric VSMC, elevation of [Ca2+]i activates a low-conductance chloride channel, which may mediate the agonist-induced depolarization of the cell membrane.
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