Enkephalin immunoreactivity and messenger RNA in a discrete projection from the nucleus of the solitary tract to the nucleus ambiguous in the rat.

ABSTRACT

Previous work described in the rat a circumscribed, partly somatostatinergic, interneuronal projection from the esophageal afferent part of the nucleus of the solitary tract (NTSc) to esophageal motor neurons in the compact formation of the nucleus ambiguous (NAcf Cunningham and Sawchenko, J Neurosci 91668, 1989). In the present study, axonal transport, immunohistochemical and in situ hybridization histochemical techniques were used to determine whether enkephalin (ENK), a peptide known to be expressed in a number of somatostatin-containing medullary cell groups, is also expressed in the projection from the NTSc to the NAcf. The results may be summarized as follows 1) cells immunoreactive (IR) for prepro-enkephalin (ppENK)-derived peptides were found in the NTSc in colchicine-pretreated animals; in untreated animals, a dense ENK-IR terminal field was observed in the NAcf sections stained with antisera against dynorphin-related peptides showed sparse staining in both regions; 2) signal indicating the presence of ppENK messenger RNA (mRNA) was found over the NTSc, including over a majority of cells identified using a retrograde tracing technique as projecting to the region of the NAcf; the signal for ppENK mRNA signal was greater than that for prepro-somatostatin (ppSS) in the NTSc; 3) a combined anterograde tracing-immunohistochemical technique demonstrated a strong correspondence between the distribution of inputs from the NTS to the NAcf, and the distribution of endogenous ENK-IR varicosities; in addition, leucine (L)-ENK-IR was found in an appreciable number of varicosities in the NAcf that had been anterogradely labeled from the NTSc; 4) unilateral electrolytic lesions of the rostromedial NTS, which included the central subnucleus, virtually eliminated ENK-IR in the ipsilateral NAcf, while staining on the contralateral side was unaffected. Taken together, these studies provide evidence that ppENK- and ppSS-derived peptides are expressed in the pathway from the NTSc to the NAcf, a pathway thought to play a role in the reflex control of esophageal peristalsis.