Toxic zinc influx may contribute to selective neuronal death after transient global ischemia. We previously used the high-affinity (K(D) = 27 nm) fluorescent dye mag-fura-5 to detect initial increases in neuronal intracellular free Zn(2+) ([Zn(2+)](i)) associated with brief Zn(2+) exposure. Here we used the specific low-affinity Zn(2+) indicator Newport Green (K(D) = 1 microm) to measure the peak levels of [Zn(2+)](i) attained during prolonged, toxic exposures to extracellular Zn(2+). Murine cortical cell cultures exposed for 5-10 min to 300 microm Zn(2+) in the presence of kainate or elevated extracellular K(+) developed widespread neuronal death over the next 24 hr. Such Zn(2+) exposure under depolarizing conditions was accompanied by a large increase in [Zn(2+)](i) reaching several hundred nanomolar, which gradually recovered over the next 20-40 min after termination of Zn(2+) exposure. Both the level of [Zn(2+)](i) elevation and the extent of subsequent neuronal death depended on the concentration of extracellular Zn(2+) between 30 microm and 1 mm. In contrast, exposure to 300 microm Zn(2+) in the presence of 300 microm NMDA resulted in little increase in [Zn(2+)](i) and little neuronal death, suggesting that NMDA receptor-gated channels are less important as a route of toxic Zn(2+) entry than voltage-gated calcium channels.