OBJECTIVE The mechanism by which apraclonidine, an alpha 2-adrenergic agonist, lowers intraocular pressure (IOP) was evaluated in humans. METHODS In a randomized, double-masked, placebo-controlled study, 0.5% apraclonidine was given topically twice daily for 1 week to one eye in each of 21 ocular hypertensive volunteers. The other eye was treated similarly with vehicle. Before and after 1 week of treatment, aqueous flow, uveoscleral outflow, fluorophotometric outflow facility, intraocular pressure, tonographic outflow facility, episcleral venous pressure, and outflow pressure were either directly measured or mathematically calculated. Values were compared in treated versus contralateral control eyes and on baseline versus day 8 of treatment. RESULTS When compared with both contralateral control eyes and baseline day, fluorophotometric outflow facility in the apraclonidine-treated eyes increased by 0.09 to 0.10 microliter/minute/mmHg (P < 0.04), IOP decreased by 3.1 to 5.2 mmHg (P < 0.0001), and outflow pressure decreased by 3.3 to 4.2 mmHg (P < 0.0001). When compared with baseline day only, aqueous flow in the apraclonidine-treated eyes decreased by 0.3 microliter/minute (P < 0.04), and episcleral venous pressure decreased by 1.0 mmHg (P < 0.001). Episcleral venous pressure also decreased in the control eyes compared with baseline day by 1.3 mmHg (P < 0.001). When compared with contralateral control eyes only, uveoscleral outflow in the apraclonidine-treated eyes decreased by 0.47 microliter/minute (P < 0.03). Tonographic outflow facility showed no change when compared with either contralateral control eyes or baseline values. CONCLUSIONS The apraclonidine-induced reduction in intraocular pressure was associated with an increase in fluorophotometric outflow facility, decrease in aqueous flow and decrease in episcleral venous pressure compared to baseline. The lack of a significant difference in aqueous flow and episcleral venous pressure between treated and contralateral control eyes may represent a contralateral drug effect.