Pacemaker channels produce an instantaneous current

Abstract

Spontaneous rhythmic activity in mammalian heart and brain depends on pacemaker currents (Ih), which are produced by hyperpolarization-activated cyclic nucleotide-gated (HCN) channels. Here, we report that the mouse HCN2 pacemaker channel isoform also produced a large instantaneous current (Iinst(HCN2)) in addition to the well characterized, slowly activating Ih. Iinst(HCN2) was specific to expression of HCN2 on the plasma membrane and its amplitude was correlated with that of Ih. The two currents had similar reversal potentials, and both were modulated by changes in intracellular Cl- and cAMP. A mutation in the S4 domain of HCN2 (S306Q) decreased Ih but did not alter Iinst(HCN2), and instantaneous currents in cells expressing either wild type HCN2 or mutant S306Q channels were insensitive to block by Cs+. Co-expression of HCN2 with the accessory subunit, MiRP1, decreased Ih and increased Iinst(HCN2), suggesting a mechanism for modulation of both currents in vivo. These data suggest that expression of HCN channels may be accompanied by a background conductance in native tissues and are consistent with at least two open states of HCN channels: Iinst(HCN2) is produced by a Cs+-open state; hyperpolarization produces an additional Cs+-sensitive open state, which results in Ih.