The rhythmic activity observed in locomotion is generated by local neuronal networks in the spinal cord. The alternating patterns are produced by reciprocal connections between these networks. Synchronous rhythmic activity, but not alternation, can be reproduced in disinhibited networks of dissociated spinal neurons of rats. This suggests that a specific network architecture is required for pattern generation but not for rhythm generation. Here we were interested in the recruitment of neurons to produce population bursts in unstructured and minimally structured cultures of rat spinal cord grown on multielectrode arrays. We tested whether two networks, connected by a small number of axons, could be functionally separated into two units and generate more complex patterns such as alternation. In the unstructured cultures, we found that the recruitment of the neurons into bursting populations is divided into two steps: the fast recruitment of a "trigger network", consisting of intrinsically firing cells connected in networks with short delays, and slow recruitment of the rest of the network. One or several trigger networks were observed in a single culture and could account for variable patterns of propagation. In the minimally structured cultures, a functional separation between loosely connected networks was achieved. Such separation led either to an independent bursting between the networks or to synchronized bursting with long and variable delays. However, no qualitatively novel pattern such as alternation could be generated. In addition, we found that the strength of reciprocal inhibitory connections was modulated by spontaneous activity.