Background: There is increasing evidence that reduced ocular blood flow plays a role in the pathogenesis of glaucoma. In patients with normal-tension glaucoma, ocular blood flow abnormalities may be associated with dysfunction of the endothelin 1 (ET-1) regulation system. Objective: To test the hypothesis that unoprostone, a topical docosanoid, may affect ET-1-induced vasoconstriction in the human choroid. Methods: In a placebo-controlled, randomized, doublemasked, 2-way crossover design, ET-1 (2.5 ng/kg per minute for 150 minutes) was administered intravenously to 24 healthy individuals. Thirty minutes after the start of ET-1 infusion, 1 drop of unoprostone or placebo was instilled into the right eye. After another 30 minutes, 2 drops of unoprostone or placebo was topically administered. This procedure was continued and the dose was increased further until 4 drops of unoprostone or placebo was reached. Subfoveal and pulsatile choroidal blood flow were assessed using laser Doppler flowmetry and laser interferometric measurement of fundus pulsation amplitude, respectively. Results: Administration of exogenous ET-1 decreased choroidal blood flow (mean±SEM, 17%±2%; PϽ.001) and fundus pulsation amplitude (mean±SEM, 19%±2%; PϽ.001). This effect was significantly blunted when topical unoprostone was coadministered (mean ± SEM decrease in choroidal blood flow, 7%±2%; P =.04 vs placebo; mean±SEM decrease in fundus pulsation amplitude, 12%±2%; PϽ.001 vs placebo). Conclusion: There is a functional antagonism between ET-1 and topical unoprostone in the choroidal vasculature. Clinical Relevance: Our findings of a functional antagonism between ET-1 and topical unoprostone in the choroidal vasculature may be important in vascular eye diseases associated with increased ET-1.