Forebrain HCN sub(1) Channels Contribute to Hypnotic Actions of Ketamine

Background: Ketamine is a commonly used anesthetic, but the mechanistic basis for its clinically relevant actions remains to be determined. The authors previously showed that HCN sub(1) channels are inhibited by ketamine and demonstrated that global HCN sub(1) knockout mice are twofold less sensitive to hypnotic actions of ketamine. Although that work identified HCN sub(1) channels as a viable molecular target for ketamine, it did not determine the relevant neural substrate. Methods: To localize the brain region responsible for HCN sub(1)-mediated hypnotic actions of ketamine, the authors used a conditional knockout strategy to delete HCN sub(1) channels selectively in excitatory cells of the mouse forebrain. A combination of molecular, immunohistochemical, and cellular electrophysiologic approaches was used to verify conditional HCN sub(1) deletion; a loss-of-righting reflex assay served to ascertain effects of forebrain HCN sub(1) channel ablation on hypnotic actions of ketamine. Results: In conditional knockout mice, HCN sub(1) channels were selectively deleted in cortex and hippocampus, with expression retained in cerebellum. In cortical pyramidal neurons from forebrain-selective HCN sub(1) knockout mice, effects of ketamine on HCN sub(1)-dependent membrane properties were absent; notably, ketamine was unable to evoke membrane hyperpolarization or enhance synaptic inputs. Finally, the EC sub(50) for ketamine-induced loss-of-righting reflex was shifted to significantly higher concentrations (by approximately 31%). Conclusions: These data indicate that forebrain principal cells represent a relevant neural substrate for HCN sub(1)-mediated hypnotic actions of ketamine. The authors suggest that ketamine inhibition of HCN sub(1) shifts cortical neuron electroresponsive properties to contribute to ketamine-induced hypnosis.