1. Influences of Ca2+ release from internal stores on the generation of depolarizing after-potentials (DAPs) were investigated in magnocellular neurones of rat supraoptic nucleus (SON) using whole-cell patch recording techniques in brain slices. 2. DAPs were recorded from more than half of the cells encountered, and following evoked single spikes had an amplitude of 3.00 +/- 0.19 mV (mean +/- S.E.M.) and lasted for 1.02 +/- 0.06 s. Their sizes usually increased with the number of preceding spikes, but could be reduced or eliminated when intervals between consecutive current pulses evoking tens of spikes were short. 3. DAPs were eliminated by removal of external Ca2+, and significantly reduced by bath application of nifedipine or omega-conotoxin. 4. Blockade of Ca2+ release from internal stores by perifusion with ryanodine or dantrolene, or direct diffusion of Ruthenium Red into cells suppressed DAP amplitudes by approximately 50% and shortened their durations. 5. Depletion of internal Ca2+ stores by perifusion with thapsigargin or cyclopiazonic acid also reduced DAP amplitudes by approximately 50% and eliminated phasic patterns of firing. 6. Caffeine, an agent known to enhance intracellular Ca2+ release, amplified DAPs and promoted phasic firing. 7. These results suggest that Ca2+ influx via high-voltage-activated Ca2+ channels in SON cells triggers ryanodine receptor-mediated Ca2+ release from internal stores. This process enhances DAPs and promotes phasic firing in SON cells, and would thus contribute to vasopressin release.