Characterization and function of Ca(2+)-activated K+ channels in arteriolar muscle cells.

We examined the functional role of large-conductance Ca(2+)-activated K+ (KCa) channels in the hamster cremasteric microcirculation by intravital videomicroscopy and characterized the single-channel properties of these channels in inside-out patches of membrane from enzymatically isolated cremasteric arteriolar muscle cells. In second-order (39 +/- 1 microns, n = 8) and third-order (19 +/- 2 microns, n = 8) cremasteric arterioles with substantial resting tone, superfusion with the KCa channel antagonists tetraethylammonium (TEA, 1 mM) or iberiotoxin (IBTX, 100 nM) had no significant effect on resting diameters (P > 0.05). However, TEA potentiated O2-induced arteriolar constriction in vivo, and IBTX enhanced norepinephrine-induced contraction of cremasteric arteriolar muscle cells in vitro. Patch-clamp studies revealed unitary K(+)-selective and IBTX-sensitive currents with a single-channel conductance of 240 +/- 2 pS between -60 and 60 mV (n = 7 patches) in a symmetrical 140 mM K+ gradient. The free Ca2+ concentration ([Ca2+]) for half-maximal channel activation was 44 +/- 3, 20 +/- 1, 6 +/- 0.4, and 3 +/- 0.5 microM at membrane potentials of -60, -30, +30, and +60 mV, respectively (n = 5), with a Hill coefficient of 1.9 +/- 0.2. Channel activity increased e-fold for a 16 +/- 1 mV (n = 6) depolarization. The plot of log[Ca2+] vs. voltage for half-maximal activation (V1/2) was linear (r2 = 0.9843, n = 6); the change in V1/2 for a 10-fold change in [Ca2+] was 84 +/- 5 mV, and the [Ca2+] for half-maximal activation at 0 mV (Ca0; the Ca2+ set point) was 9 microM. Thus, in vivo, KCa channels are silent in cremasteric arterioles at rest but can be recruited during vasoconstriction. We propose that the high Ca0 is responsible for the apparent lack of activity of these channels in resting cremasteric arterioles, and we suggest that this may result from expression of unique KCa channels in the microcirculation.