The effects of neonatal nicotine exposure on spontaneous and nicotine-induced behaviour in 4-month-old mice and on the development of brain nicotinic receptors were studied. The behaviour study showed that mice treated with nicotine 66 micrograms (-)nicotine base/kg body weight (bw) s.c. twice daily between 10 and 16 days postnatally displayed a hypoactive condition, whereas mice treated with saline displayed a hyperactive condition. When the nicotinic receptors in the brain cortex were analyzed, the displacement curves for [3H]nicotine(-)nicotine revealed an almost equal proportion of high- and low-affinity binding sites in 17-day-old mice, while the high-affinity sites predominated in 4-month-old mice, with affinity constants for both high- and low-affinity binding sites 10 times higher in 4-month-old mice than in 17-day-old-mice. A decrease in the number of nicotinic receptors was observed from day 17 to 4 months, mainly of the low-affinity nicotinic type. Interestingly, the displacement curves in neonatally nicotine-treated mice showed only one population of high-affinity binding sites in 17-day- and 4-month-old mice though the total binding sites in 4-month-old mice were the same for the neonatally nicotine-treated and saline-treated mice. These results indicate that neonatal nicotine treatment prevents the development of low-affinity nicotinic sites in the brain and this earlier exposure to nicotine induces a different behaviour response in adult animals to a test dose of nicotine. Days 10-16 postnatally appear to be a critical period for the effects of nicotine on the brain.