from Sigma-Aldrich. Whole-cell patch-clamp currents were recorded using Axopatch 200B (Molecular Devices; Axon Instruments) amplifier. Currents were adjusted electronically for cell capacitance and series resistance (80−100%), filtered at 1kHz with an 8-pole Bessel filter and digitized at 5kHz with a Digidata 1200 interface (Molecular Devices: Axon Instruments). Currents were elicited with voltage ramp protocol, from −100 mV to +50 mV, delivered at 0.5 Hz. Currents were measured at −100 mV and converted to current density (pApF−1) by dividing by the membrane capacitance. Basal K+ currents (Ba++-sensitive) were quantified at −100 mV by applying 1 mM BaCl2 in 20K and measuring the amplitude of the Ba++-inhibited current. Alcohol- and carbachol-modulated currents were measured at −100 mV by averaging current from two consecutive sweeps upon reaching steady state, and subtracting the mean basal current before and after the application of the modulator. Pooled data are presented as mean ± s.e.m. and evaluated for statistical significance (P < 0.05) using a one-way ANOVA, followed by Bonferroni multiple comparison post-hoc test. To determine the IC50, the inhibition curves were fit with the Hill equation y=1/(1+[x/c]^b), where y is fraction current remaining, x is the concentration of alcohol, b is the Hill coefficient and c
