Objectives-To investigate the effect of esophageal mechanosensitive and chemosensitive stimulation on the magnitude and recruitment of peristaltic reflexes and upper esophageal sphincter (UES)-contractile reflex in premature infants.Study design-Esophageal manometry and provocation testing were performed in the same 18 neonates at 33 and 36 weeks postmenstrual age (PMA). Mechanoreceptor and chemoreceptor stimulation were performed using graded volumes of air, water, and apple juice (pH 3.7), respectively. The frequency and magnitude of the resulting esophago-deglutition response (EDR) or secondary peristalsis (SP), and esophago-UES-contractile reflex (EUCR) were quantified.Results-Threshold volumes to evoke EDR, SP, or EUCR were similar. The recruitment and magnitude of SP and EUCR increased with volume increments of air and water in either study (P < .05). However, apple juice infusions resulted in increased recruitment of EDR in the 33 weeks group (P < .05), and SP in the 36 weeks group (P < .05). The magnitude of EUCR was also volume responsive (all media, P < .05), and significant differences between media were noted (P < .05). At maximal stimulation (1 mL, all media), sensory-motor characteristics of peristaltic and EUCR reflexes were different (P < .05) between media and groups.Conclusions-Mechano-and chemosensitive stimuli evoke volume-dependent specific peristaltic and UES reflexes at 33 and 36 weeks PMA. The recruitment and magnitude of these reflexes are dependent on the physicochemical properties of the stimuli in healthy premature infants.Gastroesophageal reflux (GER) is frequent in neonates and infants, and its role in the cause of or association with neonatal morbidity including dysphagia, chronic lung disease, or apparent life-threatening events is not understood. 1,2,3 The mechanisms of GER disease are as well defined in neonates as in adults. 4 relaxation is the most common cause, but hypotonic LES and poor esophageal peristalsis are also described. 6,7 The refluxate (gas, liquid, or mixed contents) provokes esophageal distention or acidification, and it may trigger luminal clearance to provide aerodigestive safety. Clearance mechanisms including primary peristalsis (PP), secondary peristalsis (SP) and esophago-upper esophageal sphincter (UES)-contractile reflexes (EUCR) that may prevent the entry of refluxate into the pharynx or larynx are well recognized in healthy adults and those with GER disease. [8][9][10][11] Immaturity, rapid development, and changing feeding behavior distinctly separate evolving pathophysiology in neonates from that of older subjects.Aerodigestive symptoms may result from delayed clearance, airway aspiration, or chemoreceptor stimulation. 12-15 We described methods to evaluate the esophageal body and UES motor responses resulting from esophageal provocation. [16][17][18] In our previous studies, we validated the technique of esophageal afferent provocation using different media (air or liquids) and characterized the occurrence of esophago-deglutition response (E...