The addition of nicotine decreased neuritic outgrowth in PC12 cells in culture. This effect occurs as early as one day after addition of nicotine to the culture medium in a concentration-dependent manner. The nicotine-induced decline in neurite outgrowth was prevented by d-tubocurarine (10(-4) M) indicating that the effect was mediated through a nicotinic receptor. alpha-Bungarotoxin (10(-8) M) was also able to inhibit the nicotine-induced decrease in process formation in a dose-dependent manner. The concentrations of alpha-bungarotoxin required to affect process outgrowth correlated with those required to inhibit radiolabelled alpha-bungarotoxin binding. alpha-Bungarotoxin had no effect on [3H]noradrenaline release, a functional response mediated through the alpha-bungarotoxin-insensitive neuronal nicotinic acetylcholine receptor, suggesting that alpha-bungarotoxin specifically interacts with the neuronal alpha-bungarotoxin receptor. The present results suggest a functional role for the neuronal nicotinic alpha-bungarotoxin receptor in neurite outgrowth.