Abstract: Objective To investigate the roles of hyperolarization-actived cyclic nucleotide-gated channels 1 (HCN1) in antidepressant actions of ketamine (KET). Methods Male HCN1 knock out (HCN1^-/-) and wildtype (HCN1^+/+) C57BL6 mice (8-12 weeks, 20-25 g) were chosen. The depression model of mice was developed by continuously oral administration of low dosage of corticosterone (CORT). The immobility time in forced swimming tests (FST) was used to assess the depressive state of mice. Then the two genotype depressive mice were treated with single intraperitoneal injection of 5 mg/kg ketamine (KET group, n=7) or same volume of normal saline (NS group, n=7) respectively. After treatment, the immobility time at 30 min, 24 h and 7 d after the intraperitoneal injection of ketamine or normal saline in CORT-treated mice were compared. In addition, normal HCN1^-/- and HCN1^+/+ mice were intraperitoneally injected of BrdU and then treated with 5 mg/kg ketamine (KET group, n=5) or same volume of normal saline (NS group, n=5) by single intraperitoneal injection. Each group was euthanized for immunohistochemical processing of 5-Bromo-2-deoxyuridine (BrdU)-labeled cells in hippocampus at 24 h after the intraperitoneal injection of saline or ketamine. Results The immobility time in FST of HCN1^-/- mice was less than the HCN1^+/+ mice before administration of CORT. It shows that the depressive state of HCN1^-/- mice is less intensive than that of HCN1^+/+ mice. And the immobility time in both HCN1^-/- and HCN1^+/+ mice was increased after oral administration of low dose corticosterone, with an increase in depression. In addition, the comparisons were also made to the reduction of immobility time within 30 min, 24 h and 7 d. At any time point, the CM(155.3mmreduction of immobility time in HCN1^+/+KET group was higher than those in the other three groups (P<0.05).Furthermore, there were no statistical significances among the three groups including HCN1^-/-KET group, HCN1^+/+ NS group, HCN1^-/-NS group at any point. The number of newborn neurons were more in HCN1^-/- mice than HCN1^+/+ mice after the treatment of normal saline. Compared with the NS group, the number of neonatal neurons labeled by BrdU were increased after the intraperitoneal injection of ketamine in HCN1^+/+ mice but not in HCN1^-/- mice. Conclusion Inhibition of HCN1 channels by ketamine accounts for its antidepressant actions.