1. Cultured hippocampal neurons were recorded with a patch pipette containing 100 microM of the calcium indicator Fluo-3, and one of their dendrites, carrying dendritic spines, was visualized with a x100, 1.3-numerical aperture oil objective. Calcium spikes evoked by depolarizing the somata and changes in free dendrite and spine calcium concentrations ([Ca]d and [Ca]s, respectively) were monitored with a cooled charge-coupled device (CCD) camera, acquiring images at a rate of 17-20 ms per frame. In the majority of spine-dendrite pairs, [Ca]s rose faster and to a higher level than the adjacent [Ca]d. Likewise, topical application of glutamate evoked a faster and larger change in [Ca]s than in [Ca]d. The rise of intracellular calcium concentration in response to a depolarizing current pulse, but not in response to glutamate, was reduced in the presence of the calcium antagonist verapamil in both dendrites and spines. It is suggested that dendritic spines possess voltage-gated calcium channels.