Throughout the body, mechanical perturbations are transduced into neurochemical signals by the release of ATP from non-neuronal cells. As an increased intraocular pressure (IOP) can initiate mechanical changes, we hypothesized that extracellular levels of ATP might be increased in the anterior chamber of human patients with primary acute angle closure glaucoma (PAACG). ATP levels in aqueous humor samples were determined using the luciferin-luciferase assay. Samples were obtained from 18 controls scheduled for cataract extraction and from 14 subjects with angle closure glaucoma during paracentesis. All subjects with angle closure glaucoma had remarkably elevated levels of ATP in their aqueous humor. The mean ATP concentration was ninefold higher for patients with angle closure glaucoma than for control. The concentration of ATP was correlated with IOP levels, further supporting a causal relationship. ATP levels were not influenced by the duration of the PAACG attack, suggesting the rate of ATP release was sustained. We conclude that increased IOP leads to increased levels of extracellular ATP in the anterior chamber. This elevated ATP may contribute to the extreme pain experienced by subjects with angle closure glaucoma, and may serve as a source of elevated adenosine in the anterior chamber.