Modulatory effects of genetic vs. pharmacological HCN4 channel inhibition on stimuli transmission during acute pain

Acute pain processing emerges from complex interactions among multiple brain regions, with local ion channels critically shaping neuronal communication. To better understand the role of HCN4 channels during acute pain in mice, a genetic brain-specific HCN4-KO was compared with pharmacological inhibition by the selective HCN4 channel blocker EC18. Stimulus-driven BOLD-fMRI measurements using graded peripheral thermal stimulation allowed brain-wide investigation of both discriminative and suppressive processes within ascending and descending pain pathways. Classical BOLD parameters and graph-theoretical analyses revealed that compared to controls, HCN4-KO showed a significant increase in brain activity in regions responsible for discriminative tasks, emotional pain processing and pain suppression including sensory cortex, amygdala and hypothalamus across both high and low thermal stimulation intensities. In striking contrast, acute inhibition of HCN4 with EC18 decreased activity in these same regions compared with both KO and control mice. Furthermore, comparing pre- and post-stimulation resting-state measurements revealed that HCN4-KO and controls exhibited a stimulation-induced increase in functional connectivity, whereas EC18-treated mice demonstrated a connectivity decrease. Taken together, genetic loss of HCN4 produced a hypersensitive phenotype in thermal pain processing, whereas acute pharmacological inhibition of the channel elicited an opposing hyposensitive phenotype.