In mammals, nicotinic acetylcholine receptors (nAChRs) play a crucial role in motor control. Muscle-type nAChRs mediate synaptic excitation of skeletal muscle by motoneurons, and nAChRs are present on Renshaw cells, where they produce recurrent inhibition of spinal motoneurons. We asked whether nAChRs are also present in motoneurons. Whole-cell recordings were performed on various motor nuclei in brainstem slices of young rats. Neurons were visualized using infrared (IR) videomicroscopy. Acetylcholine (ACh) or the nicotinic agonist, epibatidine, were delivered by pressure microinjection. Facial (VII), hypoglossal (XII) and vagal (X) motoneurons responded to ACh by generating a fast inward current. In VII motoneurons, the ACh effect was mimicked by epibatidine, and nicotine induced a slow inward current and desensitized the ACh-evoked current. In VII and XII motoneurons, the ACh-evoked current was blocked by the nicotinic antagonist dihydro-β-erythroidine (DHβE), but was unaffected by methyllycaconitine (MLA), an α7-specific antagonist. By contrast, the ACh-induced current in X motoneurons was sensitive to MLA. Current–voltage relationships indicated that the currents mediated by either α7-containing (X) or non-α7-containing (VII, XII) nAChRs displayed inward rectification. In accordance with the electrophysiological data, autoradiography revealed that VII, X and XII nuclei of young rats contained binding sites for [3H]epibatidine; binding sites for [125I]α-bungarotoxin, a selective ligand of α7-containing nAChRs, were present in X nucleus but were almost undetectable in VII and XII nuclei. Thus, brainstem motoneurons of young rats possess functional nAChRs. They could promote fast synaptic coupling between motoneurons, and thus play a role in somatic and visceral motor functions.