Although it is generally acknowledged that pancreatic islets are under powerful vagal control, specifics of vagal pathways and their central representation in the brain stem are unclear. To define this circuitry, we combined a protocol measuring electrical vagal stimulation-induced insulin and glucagon secretion with a retrograde tracer strategy that delineated the pool of spared motoneurons in the dorsal motor nucleus of the vagus (dmnX) following selective abdominal branch vagotomies. Three of the five branches mediated both insulin and glucagon release: posterior gastric (+198 and +117% increase from basal for insulin and glucagon, respectively), anterior gastric (+177 and +104%), and hepatic branch (+103 and +60%). In contrast, unreliable and nonsignificant hormonal responses were produced by stimulation of fibers projecting through either the posterior celiac (+12% insulin and +12% glucagon) or accessory celiac (+15% insulin and +31% glucagon) branches. Since hexamethonium almost completely blocked both insulin and glucagon responses to stimulation, the effects are not likely to have resulted from inadvertent antidromic excitation of vagal afferents. Cell bodies of stimulated motoneurons, which were responsible for insulin and glucagon secretion, were found to occupy the medial two-thirds of the right (projecting through the posterior gastric branch) and left (projecting through the anterior gastric and hepatic branches) dmnX. These medial, longitudinal dmnX columns and their associated abdominal vagal branches are likely to play the predominant role in vagal control of the endocrine pancreas.