There is evidence for a pathway involving small intestinal CCK-producing entero-endocrine cells and visceral afferent nerve fibers in signaling the effect of luminal nutrients on gastrointestinal and food intake regulation. In order to investigate the type of anatomical apposition that exists between CCK cells and vagal afferents, CCK immunocytochemistry was performed on tissue from rats whose vagal afferent fibers to the abdomen had previously been labeled in vivo by injecting the fluorescent carbocyanine dye DiI into the nodose ganglia. CCK immunoreactive (CCK-IR) cells were more abundant than vagal afferent fibers, but both were present throughout the small intestine as well as in crypts and villi. Few CCK-IR cells were in close (< 5 microns) anatomical contact with vagal afferent axons, and the latter did not produce suspicious terminal specializations near CCK-IR cells. Most labeled vagal afferent axons, which distributed strictly within the crypt and villous lamina propria, were at distances of tens to hundreds of microns to the nearest CCK-IR cell. These findings strongly support the idea that CCK released from entero-endocrine cells acts on vagal sensory fibers in a paracrine fashion, but do not rule out the presence of a few very close, neurocrine-like contacts or a humoral mode of action. Possible implications of such an arrangement on CCK-mediated satiety are discussed.