The levels of TRP, 5-HTP, 5-HT and 5-HIAA were measured in the proximal colon of the rabbit using ion exchange chromatography and spectrofluorimetry. Assays were performed on fragments of the intact colonic wall, on the muscle layers containing nerve plexuses, and on the mucosa-containing enterochromaffin cells. In spite of identical TRP concentration, 5-HT levels were higher in the mucosa than in the muscle containing nerve plexus. In the muscle, the ratio 5-HIAA/5-HT was about 1, i.e., comparable to that of the brain, whereas the ratio was very low in the mucosa, suggesting a weak 5-HT catabolism in the enterochromaffin cells. When the mucosa was severed from the muscle, 5-HT synthesis was increased. In this case, 5-HT and 5-HIAA concentrations in the reconstituted wall (muscles + mucosa) were higher than in the intact wall. In in vitro studies, levels of newly synthesized (3H)5-HT from (3H)TRP were 3 times higher in the reconstituted wall than in the intact one, and the conversion index of TRP to 5-HT was higher in the mucosa and in the reconstituted wall respectively. These results suggest that enterochromaffin cells do synthesize 5-HT from TRP when the mucosa is divided from the muscle. (3H)5-HT was taken up by both nerve plexuses and mucosa, with a high affinity mechanism. Results from tryptophan loading experiments indicated that 5-HTP decarboxylase in the gut might be involved in the regulation of 5-HT synthesis. The mucosa severed from the muscles might represent a good model for the study of carcinoid tumour development.