The cytoplasmic concentration of free calcium was measured using aequorin, a calcium-sensitive photoprotein. The Ca2+ ionophore A23187 induced a rise in cytoplasmic free calcium and iodide discharge in cultured porcine thyroid cells. The minimum dose of A23187 effecting an increase in cytoplasmic free calcium induced iodide discharge. The A23187-induced rise in cytoplasmic free calcium was followed by iodide discharge. The results indicate that A23187-induced iodide discharge is mediated by a rise in the cytoplasmic concentration of free calcium.