Twice daily topical application of 50 micrograms of prostaglandin F2 alpha tromethamine to cynomolgus monkey eyes produced significant ocular hypotension lasting at least six hours, with the intraocular pressure (IOP) falling between 35% and 50%, ie, to about 8 to 10 mm Hg, following the seventh dose. A single topical application of 1 mg of pilocarpine hydrochloride produced a much smaller IOP reduction and strong, probably maximal accommodation, both of which lasted at least eight hours. When prostaglandin F2 alpha-treated eyes were given pilocarpine before the seventh dose of prostaglandin F2 alpha, accommodation and IOP responded as in eyes receiving pilocarpine only. Atropine sulfate pretreatment of eyes receiving pilocarpine and prostaglandin F2 alpha completely prevented pilocarpine-induced accommodation and inhibition of ocular hypotension induced by prostaglandin F2 alpha. We hypothesize that (1) prostaglandin F2 alpha reduces IOP by increasing uveoscleral drainage of aqueous humor, and (2) pilocarpine pretreatment contracts the ciliary muscle, obliterating the intramuscular spaces and closing off the uveoscleral drainage pathway and thus physiologically blocking the effect.