In these studies we examined the role of excitatory amino acids (EAAs) neurotransmission in communicating sensory inputs to the airway-related vagal preganglionic neurons, by examining the effects of either NMDA or AMPA/kainate receptor blockade on reflex and chemical responses of tracheal smooth muscle. Experiments were performed in chloralose anesthetized, paralyzed and mechanically ventilated beagle dogs (n = 18), under hyperoxic, normocapnic, and normohydric conditions. Topical application or microinjection of NMDA receptor blockers, into the region of the ventrolateral medulla where airway-related vagal preganglionic neurons are located, insignificantly decreased the reflex changes in tracheal tone. However, topical application or microinjection of AMPA/kainate subtype of glutamate receptor selective antagonists markedly reduced reflex increase in tracheal tone induced by (1) lung deflation, (2) stimulation of laryngeal cold receptors, and (3) activation of peripheral or central chemoreceptors. These effects were potentiated by prior NMDA receptor blockade. Findings indicate that an increase in central cholinergic outflow to the airways by a variety of excitatory afferent inputs is mediated via activation of EAA receptors, mainly AMPA/kainate subtype of glutamate receptors.