Studies of mammalian locomotion have been greatly facilitated by the use of the isolated rodent spinal cord preparation that retains the locomotor circuits needed to execute the movement. Physiological and molecular genetic experiments in this preparation have started to unravel the basic circuit organization responsible for walking in mammals. Here, we review these experiments with a focus on the functional role of excitatory V2a interneurons in the mammalian locomotor network. With regard to these neurons and other network structures we also discuss similarities and differences between the mammalian walking central pattern generator (CPG) and the fish swimming CPG.