The parasympathetic secretomotor innervation of the salivary glands originates from a longitudinal column of neurons in the medulla called the salivatory nucleus. The neurons innervating the parotid and von Ebner salivary glands are situated in the caudal extremity of the column designated as the inferior salivatory nucleus (ISN). Immunocytochemical investigations have demonstrated the presence of a number of neuropeptides surrounding the ISN neurons. We have examined the neurophysiological effect of two of these neuropeptides on neurons of the ISN identified by retrograde transport of a fluorescent label. Both serotonin (5-HT) and substance P (SP) excited virtually all neurons in the ISN. Application of these neuropeptides resulted in membrane depolarization that was concentration dependent. While the majority of ISN neurons that were depolarized by SP application exhibited an increase in input resistance, application of 5-HT induced widely varied change in input resistance. Membrane depolarization elicited action potential discharges which increased in frequency with increasing concentration of 5-HT and SP. Blocking action potential conduction from surrounding neurons did not eliminate the depolarizing effects of 5-HT and SP indicating that both neuropeptides acted directly on the ISN neurons. Finally, the use of 5-HT agonists and antagonists indicates that 5-HT acts via a 5-HT_2A receptor, and the use of SP agonists suggests that SP acts via neuokinin-1 and neurokinin-2 receptors. These data show that 5-HT and SP excite most of the ISN neurons innervating the lingual von Ebner glands possibly modulating the synaptic drive to these neurons derived from afferent gustatory input.