Application of acetylcholine to Xenopus oocytes evoked increases in the cytosolic free calcium ion concentration ([Ca2+]i) after latencies of up to several seconds depending on the agonist dose. Higher acetylcholine concentrations evoked responses with larger amplitudes and shorter latencies. The latencies of responses to acetylcholine could be increased by application of caffeine, injection of calcium buffers or depletion of intracellular calcium stores. Acute inhibition of endoplasmic reticulum calcium pumps without substantial reduction of the calcium store content (by application of thapsigargin shortly before agonist stimulation) reduced the latencies of responses to acetylcholine. A schematic and mathematical model are presented to show a possible mechanism by which a calcium signal is initiated following a latent period after the elevation of the inositol trisphosphate concentration. During the latent period, calcium is slowly released from the intracellular stores. The released calcium is rapidly buffered by cytosolic calcium-binding proteins and some is resequestered into the stores by calcium pumps. The [Ca2+]i changes very little until the buffering is locally saturated. The [Ca2+]i then rises above a threshold concentration which evokes an explosive release of calcium due to positive feedback by calcium on the inositol trisphosphate receptor.