Effects of organotypic culture on parasympathetic innervation of guinea pig trachealis

B. J. Canning, B. J. Undem, P. C. Karakousis and R. D. Dey Johns Hopkins Asthma and Allergy Center, Baltimore, Maryland 21224, USA. Nonadrenergic, noncholinergic (NANC) relaxations of airway smooth muscle are thought to be mediated by vasoactive intestinal peptide (VIP) and nitric oxide (NO). Previous studies of the parasympathetic innervation of guinea pig trachealis suggest that the ganglion neurons mediating NANC relaxations but not cholinergic contractions are associated with the esophagus. In this study, the location of the neurons mediating these responses and their neurochemical phenotype was further assessed. Guinea pig tracheas maintained in organotypic culture for 2 days with the adjacent esophagus intact displayed cholinergic contractions and NANC relaxations to electrical field stimulation (EFS) as well as VIP and NO synthase (NOS) nerve fiber densities that were similar to those of control tracheas. By contrast, in tracheas cultured without the esophagus, NANC relaxations to EFS were not observed, and VIP and NOS nerve fiber densities were reduced > 80%. EFS-induced cholinergic contractions were unaffected by esophagus removal. These results provide further evidence that NANC relaxations are mediated by VIP and NO coreleased from noncholinergic parasympathetic nerve endings derived from neurons intrinsic to the esophagus.