Nicotine affects functions of the central nervous system. A previous study showed that developing cerebellar Purkinje cells are targets for early postnatal nicotine exposure. In this study, we assessed the effects of long-term nicotine exposure on mature cerebellar Purkinje cells. This is particularly relevant since the majority of smokers are exposed to nicotine over a long period. Female adult Sprague-Dawley rats received three doses of nicotine (0.01%, 0.03%, or 0.06%) through their sole water source. After 8 weeks of nicotine exposure, the cerebellar vermis was removed and processed for stereological cell counting. The results showed that this long-term nicotine treatment did not change the cerebellum weight or the size (volume) of the cerebellar vermis. The long-term nicotine treatment regimen did result in a significant loss of mature Purkinje cells in the cerebellum, however, such a loss of Purkinje cells was not nicotine dose-related. These findings indicated that the mature adult cerebellum is susceptible to the damaging effects of nicotine in depleting Purkinje cells in the cerebellum.