The contributions of the hyperpolarization-activated current, I_h, to generation of rhythmic activities are well-described for various central neurons, particularly in thalamocortical circuits. In the present study, we investigated effects of a general anesthetic, propofol, on native I_h in neurons of thalamus and cortex and on the corresponding cloned HCN channel subunits. Whole-cell voltage clamp recordings from mouse brain slices identified neuronal I_h currents with fast activation kinetics in neocortical pyramidal neurons and with slower kinetics in thalamocortical relay cells. Propofol inhibited the fast-activating I_h in cortical neurons at a clinically relevant concentration (5 µM); inhibition of I_h involved a hyperpolarizing shift in half-activation voltage (V_1/2 ~-9 mV) and a decrease in maximal available current (~36% inhibition, measured at -120 mV). With the slower form of I_h expressed in thalamocortical neurons, propofol had no effect on current activation or amplitude. In heterologous expression systems, 5 µM propofol caused a large shift in V_1/2 and decrease in current amplitude in homomeric HCN1 and linked heteromeric HCN1-HCN2 channels, both of which activate with fast kinetics, but did not affect V_1/2 or current amplitude of slowly-activating homomeric HCN2 channels. With GABA_A and glycine receptor-channels blocked, propofol caused membrane hyperpolarization and suppressed action potential discharge in cortical neurons; these effects were occluded by the I_h blocker, ZD-7288. In summary, these data indicate that propofol selectively inhibits HCN channels containing HCN1 subunits, such as those that mediate I_h in cortical pyramidal neurons - and they suggest that anesthetic actions of propofol may involve inhibition of cortical neurons and perhaps other HCN1-expressing cells.