Concurrent and sequential dual immunohistochemical labeling methods were used in combination, along with retrograde tracing techniques, to determine the extent to which inhibin beta (I beta), somatostatin-28 (SS-28), and enkephalin (ENK) immunoreactivity (IR) might be jointly expressed in neurons centered in the caudal part of the nucleus of the solitary tract (NTS) that project to the paraventricular nucleus of the hypothalamus (PVH). The results indicate that at least 65% of I beta-stained neurons in the NTS also express SS-28 IR, and at least 33% are ENK-positive. At least 25% of the I beta IR population stains positively for all three peptides. A substantial number of cells stained with markers for two, or all three, peptide families, could be retrogradely labeled following tracer deposits centered in the PVH. Prominent I beta and SS-28 IR projections from the caudal medulla to the hypothalamus have been described and include a preferential input to oxytocinergic (OT) compartments of the magnocellular neurosecretory system. The present results suggest that these arise in large measure from a common pool of neurons, a subset of which also shows ENK IR. Implications for the control of OT secretion, and for the processing of sensory information through the NTS, are discussed.