The projection from the limbic system via the subpallidal region to the mesencephalic locomotor region is implicated in limbic-motor integration. The goal of this study was to visualize neurons of the mesencephalic locomotor region which are active during locomotor activity induced by the disinhibition of the subpallidal region. The subpallidal region was disinhibited by picrotoxin, which antagonizes the effects of GABA. The unilateral injection of picrotoxin into the subpallidal region caused a significant increase in locomotor activity. Active tegmental neurons were subsequently visualized by immunocytochemical staining of c-Fos protein. There were significantly more immunostained neurons in the picrotoxin-injected animals than in the saline-treated rats. Heavily stained neuronal nuclei, prevailing on the brain side ipsilateral to the injection of picrotoxin, were localized within a narrow strip of tissue which stretched from the ventrolateral periaqueductal gray (including the dorsal raphe), the cuneiform nucleus, through the region of the dorsal tegmental bundle to the pedunculopontine nucleus. There were 3.5 times more immunostained neurons in the cuneiform/pedunculopontine region and 2.5 times more stained neurons in the periaqueductal region of the picrotoxin-injected rats, as compared to the saline group. This strip of immunostained cells represents neurons which are involved in the initiation and maintenance of locomotor activity due to subpallidal activation (predominantly pedunculopontine and cuneiform nuclei), as well as neurons possibly involved in the inhibition of locomotor activity (ventrolateral periaqueductal gray) and other feedback regulations. This study will help identify the neuronal pool involved in coupling the motivational commands with the locomotor system for execution of behaviour.