Characterization of vagal input to the rat esophageal muscle

Abstract

There is recent morphological evidence for an interaction of autonomic nerve fibers and extrinsic motor nerves of the rat esophagus. The aim of the present study was to investigate a possible functional role of this autonomic innervation of vagal motor fibers on rat esophageal smooth and striated muscle function in vitro. The entire esophagus with both Nn vagi, including the Nn recurrentes, was dissected and placed in an organ bath with oxygenated Krebs–Ringer buffer. Contractile activity was measured in longitudinal direction with a force transducer. Both Nn vagi were placed on a bipolar platinum electrode 2 cm apart from the esophagus. Vagal stimulation, applied for 1 s (40 V, 0.5 ms, 20 Hz) resulted in a biphasic contractile response, which was completely blocked by tetrodotoxin (10 −6 M). The first part consisted of a tetanic striated muscle contraction, which was abolished by tubocurarin (10 −5 M) but unaffected by atropine (10 −6 M) or hexamethonium (10 −4 M). In contrast, the second part was completely abolished by hexamethonium (10 −4 M) and atropine (10 −6 M), whereas tubocurarine (10 −5 M) showed no influence, suggesting a stimulation of preganglionic nerve fibers supplying esophageal smooth muscle (muscularis mucosae). In order to characterize possible autonomic transmitters of the ENS of the esophagus, the following experiments were carried out. The magnitude of the striated muscle response was unaffected by VIP (10 −7 M), 5-HT (10 −6 M) and galanin (10 −8–10 −7 M), whereas they caused an inhibition of the smooth muscle response (VIP: −53.8±4.2%; galanin 10 −8 M: −18.5±2.2%; 10 −7 M: −40.4±2.9%; 5-HT: −78.2±2.1%). The inhibitory effects of VIP and galanin on smooth muscle were reversible by the antagonists VIP 10-28 and galanin 1-15. In the presence of the nitric oxide synthase (NOS) inhibitor l-NNA (10 −4 M), the smooth and striated muscle contraction were not significantly influenced. Exogenous application of the NO-donor DEA-NO (10 −4 M) reduced the smooth muscle contraction by −81.6±7.4%, but had no significant effect on the striated muscle contraction. Though immunohistochemical findings are highly suggestive of an nitrergic autonomic modulation of striated muscle contraction by enteric neurons, we could not demonstrate a NO-mediated action on striated muscle activity. Therefore, the physiological relevance of the immunohistochemical findings remain unclear.