Anterograde tracing and immunohistochemical characterization of potentially mechanosensitive vagal afferents in the esophagus.

Vagal mechanosensitive afferents with an important functional role in esophageal peristalsis are well known from physiological studies. It is not known whether these fibers represent a separate subpopulation among all vagal afferents projecting to the esophageal wall. A morphological and immunohistochemical description of vagal afferents was undertaken to define their possible homo- or heterogeneity. The peripheral projections of vagal afferents were anterogradely labeled by injection of wheatgerm agglutinin conjugated to horseradish peroxidase into the nodose ganglion of rats. The anterogradely transported tracer was detected by tyramide amplification in conjunction with immunohistochemistry for Ca(2+)-binding proteins recently identified in different types of mechanosensory endings. It was found that vagal afferents represented a morphologically and structurally homogeneous population projecting to the myenteric ganglia of the esophagus, where they terminated as highly branched endings. Vagal afferent terminals, however, were different in their staining intensity for calretinin and calbindin, which ranged from intense to no detectable immunofluorescence. The fluorescence intensity of Ca(2+)-binding proteins within the vagal terminating branches was graded and the average staining intensity determined of all terminating branches in the upper, middle, and lower thirds of the esophagus. The average staining intensity was highest in the upper third of the esophagus and then declined in a statistically significant manner in the middle and lower thirds. This result suggests different requirements for intracellular Ca(2+)-buffering capacities in vagal afferents depending on their position along the esophageal axis and corroborates studies reporting a segmental organization of esophageal motility. Immunohistochemical evidence of substance P (SP) in a subset of vagal terminals was demonstrated. Hence, an effector role of vagal afferents on esophageal peristalsis by the release of SP, as has been proposed by physiological studies, is also supported by immunohistochemical data.