Hyperpolarization‐activated cation currents (I_h) have been identified in cardiac pacemaker cells and a variety of central and peripheral neurons. Four members of a gene family encoding hyperpolarization‐activated, cyclic nucleotide‐gated cation channels (HCN1–4) have been cloned recently. Native I_h currents recorded from different cell types exhibit distinct activation kinetics. To determine if this diversity of I_h currents may be caused by differential expression of HCN channel isoforms, we investigated the cellular distribution of the transcripts of HCN1–4 in the murine sinoatrial node, retina and dorsal root ganglion (DRG) by in situ hybridization. In the sinoatrial node, the most prominently expressed HCN channel is HCN4, whereas HCN2 and HCN1 are detected there at moderate and low levels, respectively. Retinal photoreceptors express high levels of HCN1, whereas HCN2, 3 and 4 were not found in these cells. In DRG neurons, the dominant HCN transcript is HCN1, followed by HCN2. We next determined the functional properties of recombinant HCN1–4 channels expressed in HEK293 cells. All four channel types gave rise to I_h currents but displayed marked differences in their activation kinetics. Our results suggest that the heterogeneity of native I_h currents is generated, at least in part, by the tissue‐specific expression of HCN channel genes.