The distribution of vasopressin binding sites in the hypoglossal nucleus of newborn rats was determined using autoradiography on film and a radioiodinated vasopressor antagonist. These sites predominated in the ventromedial and dorsal divisions of the nucleus. The effect of vasopressin on hypoglossal neurones was studied in brainstem slices of newborn animals, using the single-electrode voltage-clamp technique. Vasopressin, at 0.1-0.5 microM, generated a sustained inward current in a majority of neurones, an action which was mediated by V1-type receptors. Antidromic activation or morphological characterization of biocytin-labelled neurones indicate that part of the vasopressin-sensitive cells were motoneurones. When synaptic transmission was blocked by perfusing the preparation with a low-calcium/high-magnesium solution, the average vasopressin current decreased by 65%; and following TTX treatment, the peptide current decreased by 55%. In contrast, in a low-calcium solution, i.e., under conditions of reduced synaptic transmission but of increased neuronal excitability, the vasopressin current was not significantly altered. These results may be interpreted by assuming that the action of vasopressin is in part postsynaptic and in part presynaptic, the latter effect probably depending upon action potential propagation. Current-voltage relations suggest that the postsynaptic effect of vasopressin was due to the induction of a non-inactivating inward current, reversing in polarity at around -15 mV. The data raise the possibility that, in young animals, endogenous vasopressin may modulate the activity of hypoglossal motoneurones.