The effect of nicotine on evoked glutamate release from isolated nerve terminals (synaptosomes) from rat prefrontal cortex was examined. We found that nicotine significantly potentiated 4-aminopyridine (4AP)-evoked glutamate release, and this potentiatory effect was mimicked by the selective alpha7 nicotinic receptor agonist choline and was blocked by the selective alpha7 nicotinic receptor antagonist methyllycaconitine, indicating its mediation by alpha7 nicotinic receptors. Examination of the effect of nicotine on cytosolic [Ca(2+)] revealed that the potentiation of glutamate release was associated with an increase in voltage-dependent Ca(2+) influx through N- and P/Q-type Ca(2+) channels. The potentiatory effect of nicotine on Ca(2+) influx seems to be attributed to its increasing synaptosomal excitability because nicotine significantly increased depolarization-evoked increase in the intrasynaptosomal free Na(+) concentration and 4AP-evoked depolarization of the synaptosomal plasma membrane potential. Also, Ca(2+) ionophore ionomycin-induced glutamate release was enhanced by nicotine, and this action was blocked by methyllycaconitine. These results suggest that nicotine exerts its potentiatory effect presynaptically, likely through the activation of alpha7 nicotinic receptors, resulting in Na(+) influx and local depolarization, which subsequently enhances the Ca(K+) entry through voltage-dependent N-and P/Q-type Ca(2+) channels as well as the vesicular release machinery to cause an increase in evoked glutamate release from rat prefrontocortical nerve terminals. Moreover, in this release potentiation may involve an activation of Ca(2+)/calmodulin signaling pathway as nicotine-mediated potentiation of 4AP- and ionomycin-evoked glutamate release were significantly attenuated by KN62, a selective inhibitor of Ca(2+)/calmodulin-dependent kinase II.