Acute administration of non-selective and relatively selectiveβ-adrenoceptor agonists elicit a monophasic fall in IOP. The present study indicates that unilateral application of certainβ-agonists on consecutive days can result in marked ocular hypertension. 1-Epinephrine, reproterol and 1-isoproterenol evoked an elevation of IOP on the second and third day after topical administration of a 2% solution in normal rabbits and in rabbits with surgically transected extraocular muscles. In contrast, the same concentration of d-isoproterenol produced hypotensive responses only when administered once daily for three consecutive days. Since d-isoproterenol did not cause a rise in IOP during chronic administration, the mechanism involved in the hypertensive response appears to be more sensitive to the levorotatory form. Administration of timolol inhibited the ocular hypertensive effect of epinephrine and reproterol. Since timolol lowers IOP by depressing formation of aqueous, it is suggested that the rise in IOP following chronic administration ofβ-agonists possibly involves an increase in aqueous humor production.