Background and Aims: we recently found that an orally directed stretch of the esophagus activates a neurally mediated relaxation of the lower esophageal sphincter (LES). Goals of our study were to characterize the neural mechanisms responsible for axial and transverse stretch-activated responses in the LES. Methods: LES pressure was monitored in anesthetized and artificially ventilated mice. Sutures were placed in the esophagus to exert graded stretch in the longitudinal and transverse direction. Effects of bilateral vagotomy and pharmacological agents on the stretch activated LES responses were investigated. Relationship between vagal stimulated axial stretch and LES relaxation was also studied. Results: Stretch in the longitudinal and transverse directions caused a dose dependent LES relaxation and contraction respectively that were not affected by bilateral vagotomy and sympathectomy but were blocked by Tetrodotoxin. In bilateral vagotomized animals, hexamethonium, atropine , PPADS and ondansetron did not block the stretch-activated LES relaxation and contraction. Axial stretch activated LES relaxation was blocked by nitric oxide inhibitor and transverse stretch activated LES contraction was blocked by a combination of atropine and substance P antagonist. Electrical stimulation of the vagus nerve induced LES relaxation and axial stretch on the LES, both of which were blocked by rocuronium. Axial and transverse stretch activated LES relaxation and contraction were present in the W/W^v mice that lack interstitial cells of Cajal (ICC). Conclusions: Stretch-activated LES relaxation and contraction are mediated through mechanosensitive neurons located in the myenteric plexus that neither involves synaptic transmission nor ICC.