The high incidence of smoking among alcoholics may be partially due to nicotine's ability to counteract some of the adverse effects of ethanol on motor coordination and/or cognitive functions. Neuroprotective effects of nicotine on ethanol-induced toxicity in cerebellar granular cells have been observed. In this study, we sought to determine whether similar protection is observed in neocortical cells and if so, what specific nicotinic receptor subtypes may be mediating the actions of nicotine. Primary cultures of neocortical cells were prepared from 20-day embryos obtained from time-pregnant Sprague-Dawley rats. Cells were cultured for 10 days and were then exposed for 3 days to various concentrations of ethanol with and without pretreatment with nicotine and nicotinic antagonists. Cellular toxicity was evaluated by measuring the lactate dehydrogenase level. Administration of ethanol (10-100 mM) resulted in a dose-dependent toxicity. Pretreatment with nicotine 5-20 microM resulted in a dose-dependent protection against ethanol-induced toxicity. The effects of nicotine were blocked by pretreatment with nicotinic antagonists such as mecamylamine (1-20 microM), dihydro-beta-erythroidine (DHBE) 50 nM-1.0 microM and methyllycaconitine (MLA) 5 nM-1 microM in a dose-dependent manner. Compared to previous studies, higher ethanol concentrations were required to induce toxicity in neocortical vs cerebellar granule cells. Moreover, the effects of nicotine in the neocortical cells were blocked by lower concentrations of MLA, but higher concentrations of DHBE compared to cerebellar cells. Collectively, the results suggest differential sensitivity of various neuronal populations to the toxic effect of ethanol. Furthermore, protective effects of nicotine against alcohol in various regions appear to be mediated by different nicotinic receptor subtypes. The neuroprotective effect of nicotine against ethanol-induced toxicity may be a contributing factor to the high incidence of smoking among alcoholics.