Smoking during pregnancy is associated with numerous physiological and neurobehavioral deficits in infants, which persist into adolescence. To better understand the underlying mechanisms, we have treated pregnant rats with nicotine and have evaluated expression of the immediate early gene c-fos, as a measure of neuronal activity, in the brains of adolescent male offspring. Pregnant dams were infused with nicotine (3 mg/kg/day) or saline from gestational day (G) 4 until G18. After birth on G22, litters were cross fostered and weaned at postnatal day (P) 21. Brain sections from adolescent offspring, aged P38-40, were analyzed by in situ hybridization for regional c-fos mRNA expression in response to acute injection of saline or nicotine (0.03, 0.1, 0.3 mg/kg). Acute nicotine challenge increased c-fos expression within nucleus accumbens shell, lateral bed nucleus of the stria terminalis, paraventricular nucleus of the hypothalamus, dorsal lateral geniculate, and superior colliculus, whereas c-fos expression was decreased in prelimbic cortex. There was no effect of gestational nicotine treatment on acute nicotine-induced alterations in c-fos mRNA levels. However, basal c-fos mRNA expression within infralimbic cortex and nucleus accumbens core was increased by gestational nicotine treatment. These data indicate that gestational nicotine does not produce global changes in nicotine-induced c-fos expression in adolescent brain. However, gestational drug exposure changes basal neuronal activity within mesocorticolimbic structures that are critical for motivated behavior. Such changes may underlie some of the behavioral deficits in attention, cognition, and impulse control that have been reported in the offspring of smoking mothers.