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The effect of prostaglandin E₂ (PGE₂) on the secretion of adrenaline and noradrenaline by the adrenal gland and the interaction between PGE₂ and acetylcholine in the adrenal medulla were examined in anaesthetized dogs. In splanchnicotomized dogs, i.v. injection of PGE₂ failed to induce any secretion of catecholamines from the adrenal gland, whereas administration of PGE₂ into the lumboadrenal artery resulted in a slight, approximately dose-dependent increase in catecholamine secretion within 2 min of the injection. This effect of PGE₂ was unaffected by i.v. administration of atropine. Intravenous administration of acetylcholine 1 min after the administration of PGE₂ into the lumboadrenal artery of splanchnicotomized atropine-treated dogs had a markedly greater effect on adrenal catecholamine secretion; the resultant output was about twice that evoked by acetylcholine in the absence of PGE₂. The effect was more than additive, since the response to acetylcholine was at least one order of magnitude greater than that to PGE₂. This indicates that PGE₂ and acetylcholine may act synergistically in the adrenal medulla.