1. The effects of lifarizine (RS-87476) on intracellular Ca2+ rises and the release of glutamate from rat cerebrocortical synaptosomes depolarized with 30 mM KCl were investigated by use of entrapped fura 2 and exogenous glutamate dehydrogenase. 2. Prior (1 min) addition of lifarizine decreased 30 mM KCl-induced total glutamate release, with 3 microM and 10 microM causing 39% and 72% averaged decreases from controls. The calcium-dependent component of glutamate release (approx. 40% of total) was similarly decreased by 47% and 74%, whereas the calcium-independent component was decreased by only 32% and 43% respectively. 3. In parallel experiments with fura-2-loaded synaptosomes, lifarizine reduced the depolarization-induced increases in intracellular [Ca2+], suggesting that this is the means by which the decreases in glutamate release are brought about. Lifarizine inhibited both the plateau and the spike phases of the Ca2+ increases suggesting that, in addition to its known sodium channel blocking properties, it may also inhibit more than one class of calcium channel in the synaptosomes. 4. Lifarizine at 1 microM and 3 microM also inhibited the rises in intracellular [Ca2+] in rat cultured cortical neurons depolarized with 60 mM KCl. 5. These effects of lifarizine on intracellular Ca2+ and glutamate exocytosis may contribute to its neuroprotective action.