["Acta Physiologica, Volume 231, Issue 4, April 2021. ", "\nAbstract\n\nAim\nAcetylcholine release is vital in the pacing of theta rhythms in the hippocampus. The subiculum is the output region of the hippocampus with different neuronal subtypes that generate theta oscillations during arousal and rapid eye movement sleep. The combination of intrinsic resonance in the hippocampal neurons and the periodic excitation of hippocampal excitatory and inhibitory neurons by cholinergic pathway drives theta oscillations. However, the acetylcholine mediated effect on intrinsic subthreshold resonance generating hyperpolarization‐activated cyclic nucleotide‐gated current, Ih of subicular neurons is unexplored. We studied the acetylcholine receptor‐independent effect of cholinergic agents on the intrinsic properties of subiculum principal neurons and the underlying mechanism.\n\n\nMethods\nWe bath perfused acetylcholine or nicotine on rat brain slices in the presence of synaptic blockers. The physiological effect was studied by cholinergic fibres stimulation and electrophysiological recordings under whole‐cell mode of subiculum neurons using septohippocampal sections.\n\n\nResults\nExogenously applied acetylcholine in the presence of atropine affected two groups of subicular neurons differently. Acetylcholine reduced the resonance frequency and Ih in bursting neurons, whereas these properties were unaffected in regular firing neurons. Subsequently, the endogenously released acetylcholine by stimulation showed a selective suppressive effect on Ih, sag, and resonance in burst firing among the two excitatory neurons. Nicotine suppressed the Ih amplitude in burst firing neurons, which was evident by decreased sag amplitude and resonance frequency and increased excitability.\n\n\nConclusion\nOur study suggests cell type‐specific acetylcholine receptor‐independent shift in resonance frequency by partially inhibiting HCN current during high cholinergic inputs.\n\n"]