We examined the protective effect of nicotine against neuronal cell death in the model system of serum- and nerve growth factor (NGF)-free cultures of PC12 cells. Serum deprivation induced massive death of undifferentiated PC12 cells, which was inhibited by the addition of NGF, but not by the addition nicotine (100 microM). Even after PC12 cells had been differentiated by the treatment with NGF, serum and NGF deprivation induced rapid and massive death of these cells. The addition of nicotine (1-100 microM) to the deprivation system prevented such cell death almost completely. The protective effect of nicotine was abolished by hexamethonium or mecamylamine, inhibitors of nicotinic acetylcholine (nACh) receptors. Treatment with reserpine to deplete intracellular catecholamines did not influence the effect. The results suggest that the protective effect of nicotine against neuronal cell death is exerted through nACh receptors.