Gastric peristaltic contractions are driven by electrical slow waves modulated by neural and humoral inputs. Excitatory neural input comes primarily from cholinergic motor neurons, but acetylcholine causes depolarization and chronotropic effects that might disrupt the normal proximal-to-distal spread of gastric slow waves. We studied cholinergic responses in stomach tissues taken from wild-type and W/W^V mice using intracellular electrical recording techniques. Electrical field stimulation (5 Hz) enhanced slow wave frequency. These effects were abolished by atropine and the muscarinic M₃ receptor antagonist, 4-DAMP. ACh released from nerves did not depolarize antral muscles. At higher rates of stimulation (10 Hz), chronotropic effects were mediated by both ACh and a non-cholinergic transmitter and blocked by muscarinic antagonists and NK₁ and NK₂ receptor antagonists. Neostigmine enhanced slow wave frequency, suggesting that the frequency of antral pacemakers is kept low by efficient metabolism of ACh. Neostigmine had no effect on slow wave frequency in muscles of W/W^V mice, which lack intramuscular ICC (ICC-IM). These muscles also showed no significant chronotropic response to 5 Hz EFS or the cholinergic agonist carbachol. The data suggest that the chronotropic effects of cholinergic nerve stimulation occur via ICC-IM in the murine stomach. The capacity of gastric muscles to metabolize ACh released from enteric motor neurons contributes to the maintenance of the proximal-to-distal slow wave frequency gradient in the murine stomach. ICC-IM play a critical role in neural regulation of gastric motility, and ICC-IM become the dominant pacemaker cells during sustained cholinergic drive.