A split-bath in vitro brainstem-spinal cord preparation from adult turtles (Pseudemys scripta) was used to test the hypotheses that: (1) bulbospinal respiratory synaptic transmission is mediated, at least in part, by N-methyl-D-aspartate (NMDA) glutamatergic receptors, and (2) this transmission is suppressed by low spinal pH (induced by hypercapnea). Recordings from intact pectoralis (expiratory) and serratus (inspiratory) nerves showed that the in vitro turtle brainstem-spinal cord preparation produces phasic expiratory and inspiratory bursts of activity similar to that produced by intact turtles. Bath application of AP-5 [(+/-)-2-amino-5-phosphonopentanoic acid, a noncompetitive NMDA receptor antagonist] to the spinal cord reversibly reduced pectoralis and serratus burst amplitude. In contrast, lowering the pH from 8.04 to 6.94 did not alter burst amplitude in either pectoralis or serratus nerves. These data suggest that spinal NMDA receptors: (1) mediate part of the bulbospinal respiratory synaptic transmission, and (2) are pH/P(CO2)-insensitive. The pH/P(CO2)-insensitivity of NMDA-dependent bulbospinal respiratory synaptic transmission may represent an important adaptation in turtles.