Vasoactive intestinal polypeptide (VIP) and nitric oxide synthase (NOS) positive innervation patterns were immunohistochemically and statistically evaluated in the human colon. Specimens from the right colon (cecum, ascending and right transverse colon) and left colon (left transverse and descending colon) were obtained surgically, fixed either in paraformaldehyde or in Carnoy's or in Bouin's and paraffin embedded. Sections were stained with hematoxylin-eosin, toluidine blue, cresyl violet, neuron-specific enolase, anti-VIP, and anti-NOS. The same results were obtained regardless of the fixative used. Enolase-positive, VIP-positive, and NOS-positive cells were occasionally found within the circular muscle and interpreted as neurons. VIP-positive nerve fibers were evenly distributed within the circular muscle while NOS-positive ones were lacking in its inner portion. The left colon was richer in neurons than the right colon, at both plexuses. VIP- and NOS-positive neuron densities were higher at the left than at the right colon, whereas at all colonic levels VIP-positive neuron percentages at both plexuses and NOS-positive ones at the myenteric plexus were similar. At the submucous plexus the NOS-positive neuron percentage was lower than that of the VIP-positive one. CONCLUSIONS (a) the right colon contains a lower number of neurons and of VIP- and NOS-positive ones than the left colon, and (b) VIP- and NOS-positive fibers are differently distributed in the inner and outer portions of the circular muscle.