Intracellular Ca2+ was imaged in cultured neonatal rat retinal neurons using the Ca(2+)-sensitive dye fluo-3 and confocal scanning laser microscopy. Depolarization via elevation of bath K+ concentration resulted in large cytoplasmic and nuclear Ca2+ signals; responses in the nucleus exceeded those of the cytoplasm. Glutamate or kainate application elicited the same intracellular pattern of elevated Ca2+ signals. Kainate stimulation was blocked by the non-NMDA receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), and greatly reduced by removing Ca2+ from the bath and adding ethylene glycol-bis (beta-amino-ethyl ether) N,N,N',N'-tetraacetic acid (EGTA). Kainate was equally effective in eliciting Ca2+ signals when bath Na+ was replaced with equimolar concentrations of choline, or in the presence of the NMDA receptor antagonist, 2-amino-5-phosphonovaleric acid (APV). Caffeine treatment significantly reduced the kainate-induced intracellular Ca2+ response. These results suggest that Ca2+ can enter through the kainate receptor of retinal neurons and amplify the Ca2+ signals in the cytoplasm and nucleus by releasing Ca2+ from intracellular stores.