Results from several studies suggest that the ventrolateral medulla (VLM) is involved in modulating the respiratory response to central and/or peripheral chemoreceptor stimulation. Furthermore, the excitatory amino acid (EAA) glutamate has been shown to have marked effects on respiration when administered to VLM sites. The purpose of this study was to determine if an excitatory amino acid mechanism in the VLM modulates the respiratory responses to hypoxia or hypercapnia in anesthetized rats. Exposure to hypoxic or hypercapnic gas under control conditions elicited increases in respiratory activity (diaphragmatic EMG activity and breathing frequency). Bilateral injection of kynurenic acid (KYN), an EAA antagonist, into rostral VLM sites evoked significant increases in breathing frequency; injections more caudal in the VLM typically caused apnea. Significantly larger increases in respiratory output were elicited by both hypoxia and hypercapnia after rostral VLM microinjections of KYN. The accentuated responses returned to control levels after a recovery of approximately 100 min. Microinjection of xanthurenic acid (XAN), an inactive analog of kynurenic acid, into the VLM prior to KYN had only slight effects on resting respiratory activity and no effects on the responses to hypoxia or hypercapnia. These results suggest two separate VLM sites which modulate respiration by EAA mechanisms. A more rostral site tonically inhibits respiratory activity and the respiratory responses to chemoreceptor stimulation and more caudal VLM sites may be required for the maintenance of respiratory activity.