Propagation of action potentials in axons and dendrites increases intracellular Na+ ([Na+]i) and Ca2+ concentrations ([Ca2+]i). While the importance of [Ca2+]i in synaptic transmission is well established, a possible functional role of [Na+]i is unclear. In cultured hippocampal cells, [Na+]i was increased by veratridine. We have then measured spontaneous excitatory postsynaptic currents (sEPSCs) and, by means of fluorescent dyes, changes in [Na+]i, in [Ca2+]i, and in the turnover of the vesicular pool of individual boutons. An elevation of [Na+]i and a concomitant rise in [Ca2+]i, led to a large increase in sEPSC frequency and in the turnover of the presynaptic vesicular pool. Extracellular Ca2+ was essential for these effects of elevated [Na+]i on synaptic transmission. They probably occur via Na+/Ca2+ exchange.