Neurons of the medial pontine reticular formation (mPRF) are involved in the execution of numerous behaviors including initiation of locomotion, eye movements, startle responses, and rapid eye movement sleep phenomena. Approximately half of the afferent projections to mPRF neurons come from within the reticular formation (Shammah-Lagnado et al., 1987). In spite of the importance of reticulo-reticular connections, virtually nothing is known about transmitters mediating these synapses. In order to identify a candidate excitatory neurotransmitter, the actions of excitatory amino acids (EAAs) on the membrane properties of mPRF neurons recorded in rat brainstem slices in vitro were studied. Standard intracellular recording methods, including single-electrode voltage clamp, were used to examine the postsynaptic actions of EAAs. We also tested whether EAA antagonists block EPSPs evoked by stimulation of the contralateral reticular formation in the slices. mPRF neurons responded to both non-NMDA and NMDA agonists. NMDA-induced conductances were voltage dependent and depressed by physiological concentrations of magnesium. Stimulation of the contralateral reticular formation elicited EPSPs that were depressed by the general EAA antagonist kynurenate. Evoked EPSPs were partially depressed by 6,7-dinitroquinoxaline-2,3-dione. The evoked EPSP was further reduced by the NMDA antagonist (+/-)-2-amino-5-phosphonopentanoic acid in some cases. These results suggest that excitatory reticulo-reticular neurotransmission is mediated by an EAA. Both non-NMDA and NMDA receptors contribute to EAA neurotransmission in the mPRF formation and play an integral role in reticular formation function.