Effects of CCK, VIP, PACAP38, and PACAP27 on the release of 5HT into the intestinal lumen and into the portal circulation were examined in in vivo experiments of isolated rat duodenum perfused vascularly and luminally. VIP, PACAP 38 and 27 reduced the release of 5HT into the lumen but did not affect the vascular release of 5HT. These effects were not affected by the presence of atropine, hexamethonium, or TTX, suggesting that VIP, PACAP 38 and 27 exert a direct inhibitory effect on the luminal release of 5HT from the EC cells. Nitric oxide synthase inhibitor, NG-nitro-L-arginine, antagonized the inhibitory effects of VIP, PACAP 38 and 27, suggesting that nitric oxide seems to be essential to exert the inhibitory action of VIP and PACAPs on the release of 5HT into the intestinal lumen from the EC cells.