The CNS monoamine cell groups that project to the pancreatic parasympathetic preganglionic neurons were identified with the use of the viral retrograde transneuronal labeling method. Pseudorabies virus (PRV) was injected into the pancreas of C₈ spinal rats and subsequently, transneuronally-labeled central monoamine neurons were mapped in brain tissue sections that had been stained by an immunohistochemical procedure that allowed for the visualization of PRV products and biogenic amine neurotransmitter enzymes or serotonin (5-HT) in the same neuron. The enzymes studied were tyrosine hydroxylase (TH), dopamine-β-hydroxylase (DBH), phenylethanolamine-N-methyltransferase (PNMT), and histidine decar☐ylase. Pancreatic vagal motor neurons originate exclusively from the dorsal vagal motor nucleus and some of these may be dopamine neurons because they were TH immunopositive, but DBH and PNMT immunonegative. Transneuronally labeled aminergic neurons were found throughout the medulla oblongata. The adrenergic inputs arose from the C1, C2, and C3 cell groups. Noradrenergic inputs originated predominantly from the A5 cell group, with lesser contributions from the A1 and A2 cell groups as well as from the area postrema. None of the other CNS catecholamine cells were labeled, except for some weakly staining TH-immunoreactive neurons, presumably dopaminergic, in the paraventricular hypothalamic nucleus (PVN). The greatest number of 5-HT neurons that innervate the pancreatic vagal motor neurons come from the gigantocellular reticular nucleus, pars alpha with lesser inputs from the raphe magnus, obscurus, and pallidus nuclei. None of the CNS histaminergic cell groups were labeled. In conclusion, the present study demonstrates that pancreatic vagal motor neurons receive inputs from adrenergic, noradrenergic, and 5-HT neurons of the lower brainstem and from a potential dopaminergic input from the PVN.