The development of peptidergic neurons in the foregut of the chick.

We have used antisera specifically directed against Met-enkephalin (met-ENK), somatostatin (SOM), substance P (SP), and vasoactive intestinal peptide (VIP), to study the development of neurons containing these peptides in the foregut of the chick. All four peptides were detected early in ontogeny, at 4 to 9 days of incubation (d.i.), and were localized primarily to cell bodies in the primitive myenteric plexus. There were differences in the times at which they were first detected and in the sequence of their appearance in the proventriculus, gizzard, and duodenum. The differentiation of these peptidergic neurons in the duodenum was examined in some detail. Cell bodies containing these peptides were first detected in the primitive myenteric plexus at 5 to 7 d.i. and increased in number from 7 to 11 d.i. Processes containing varicosities became prominent between 11 and 13 d.i. VIP was the first of the peptides to appear in the submucosal plexus and was found in more proximal regions of the duodenum at 5 d.i. Shortly thereafter, SOM- and SP-containing cell bodies were seen; met-ENK-containing cell bodies were never detected in the submucosal plexus. At 13 d.i., the circular smooth muscle contained a number of VIP-immunoreactive and a smaller number of SOM-immunoreactive processes. met-ENK- and SP-immunoreactive processes appeared in the circular smooth muscle between 17 and 21 d.i.; VIP- and SP-immunoreactive processes appeared in the mucosal plexus at 17 to 21 d.i. Our results suggest that neuropeptides appear very early in the ontogeny of enteric neurons, at the same time or even before cholinergic and serotonergic neurons express their phenotypes. These findings argue against a sequential developmental order in which peptidergic neurons appear after those containing acetylcholine and serotonin.