Endotracheal suctioning of intubated infants produces profound changes in cardiovascular and cerebral hemodynamics, but the mechanisms regulating these changes are not fully understood. To determine the role of the autonomic nervous system in regulating these physiologic changes, we investigated the effects of endotracheal suctioning on heart rate (HR), mean arterial blood pressure (MABP), and renal sympathetic nerve activity (RSNA) in nine ventilated newborn lambs. In the first part of the study (n = 6), ventilation was interrupted for suctioning. With suctioning (15 s), HR decreased by 39 +/- 6% (p < 0.05), whereas MABP and RSNA increased significantly (p < 0.05) by 36 +/- 5% and 68 +/- 8%, respectively. These changes were significantly (p < 0.05) larger than changes observed during disconnection from the ventilator (15 s) without suctioning. Administration of atropine (0.02 mg/kg) blocked the HR response to suctioning without altering MABP or RSNA changes. After bilateral vagotomy, suctioning produced no changes in any parameter. When a closed tracheal suction system was used and ventilation was maintained, suctioning again resulted in significant (p < 0.05) increases in MABP (+10 +/- 3%) and RSNA (+34 +/- 5%) and a decrease in HR (-15 +/- 4%). These data suggest that suctioning stimulates sympathoexcitatory receptors localized in large airways whose afferent fibers course within the vagus, resulting in increased sympathetic activity, which induces peripheral vasoconstriction and elevates MABP. In contrast, the HR response appears to be mediated by increased parasympathetic activity as this is abolished by atropine.