The administration of methamphetamine (2 mg/kg SC) to mice was followed by acceleration of ambulation (locomotion) for 3 h, with the peak effect at 1/2-1 h. When mice were allowed free ambulation for 3 h in an activity cage of 20 cm in diameter after methamphetamine administration, repeated administrations of methamphetamine, four times at 3-day intervals, caused ambulatory sensitization; the count at the fourth administration being approximately 2.3 times as high as that at the first administration. Furthermore, the mice that were allowed ambulation in the activity cage during postmethamphetamine period of either 0-1/2, 0-1, 0-2, or 1/2-1 h (during the other periods, the mice were put in jars of 6 cm in diameter to restrict ambulation) demonstrated ambulatory sensitization to methamphetamine, and these levels were as same as that in the mice given methamphetamine with free ambulation for 3 h. However, the free ambulation during postmethamphetamine periods of 0-1/4 h and 1-3 h failed to produce strong sensitization to methamphetamine. These results suggest that a free ambulation for at least 1/2 h during the postmethamphetamine period of 0-1 h is important for induction of ambulatory sensitization to methamphetamine in mice.