Ambulatory activity in rats was increased in a dose-related manner by microinjections of picrotoxin, a GABA antagonist, bilaterally into the ventral tegmental area. Microinjections of strychnine, a glycine antagonist, had no effect on activity. The ambulatory activity induced by microinjections of picrotoxin into the ventral tegmental area was significantly attenuated when the nucleus accumbens was pretreated with spiroperidol, a dopamine antagonist. These findings provide additional evidence that dopaminergic (A10) neurons projecting from the ventral tegmental area to the nucleus accumbens, contribute to locomotor activity. It is suggested that picrotoxin disinhibits the A10 dopaminergic neurons projecting to the nucleus accumbens.