Locomotor stepping elicited by lateral hypothalamic stimulation in the anesthetized rat is blocked by lesions in the anterior ventromedial midbrain. This study determined in acute experiments whether the dorsal midbrain regions implicated in locomotion were also part of the necessary pathway. Rats were anesthetized with Nembutal and held in a stereotaxic apparatus so that stepping responses rotated a wheel. Stepping was elicited by stimulation of the lateral hypothalamus (up to 100 microA, 0.5 ms cathodal pulses, 50 Hz, 10-s train length). Nine rats received unilateral lesions ipsilateral to the locomotor electrode and 3 rats received bilateral lesions. None of the dorsal midbrain lesions reduced locomotion elicited by ipsilateral lateral hypothalamic stimulation. Therefore the following regions are unnecessary for this type of locomotion: the dorsal and lateral central gray, the tegmentum lateral to the central gray, and in particular the area cuneiformis and the dorsal aspect of the pedunculopontine region. The neural systems required for lateral hypothalamic locomotion are located ventral to the superior cerebellar peduncle.