The development of motor networks in the spinal cord is partly activity-dependent. We have observed receptor-mediated excitatory effects of two peptides, arginine vasopressin (AVP) and oxytocin (OXT), on motor network activity in the neonate. With the use of an en bloc in vitro preparation of mouse spinal cord (2-3 d old), which either was isolated completely or had muscles of the hindlimb left intact, we show that the bath application of AVP or OXT can evoke an increase in population bursting of motoneurons recorded from the lumbar ventral roots. By using antagonists for AVP and OXT, we found that these peptides were binding primarily to V1a and OXT receptors, respectively. Western blot analysis revealed a 48 kDa V1a and a 55 kDa OXT receptor immunoreactive band that was expressed in tissue obtained from L1-L6 sections of spinal cord. AVP, but not OXT, could, on occasion, evoke sustained periods of locomotor-like activity. In addition, when we applied AVP or OXT in combination with a 5-HT2 agonist, bouts of locomotor-like activity could be observed in a majority of preparations. Collectively, these data point to a novel role for AVP and OXT in the activation of spinal motor networks.