The tachykinin substance P (SP) is present in the ventral and medial area of the lumbar spinal cord. Its localisation suggests that it could modulate the spinal network for locomotion. We have investigated its effects on motor outputs by applying SP, in vitro, to the lumbosacral segments of an isolated spinal cord of new-born rats. SP was applied to the lumbosacral spinal cord either on a quiescent preparation or during episodes of fictive locomotion induced by N-methyl-D,L-aspartate. When applied on quiescent preparations, SP induced a slow rhythmic activity (period >30 s). During fictive locomotion, SP increased both the locomotor frequency and the duration of the bursts of cyclic activity. Furthermore, SP stabilised the locomotor rhythm. These results demonstrate that SP is able to modulate both the "clock" and the pattern generator for locomotion.