Contraction of the ciliary muscle induced by cholinergic drugs causes movement of the scleral spur and separation of the trabecular meshwork lamellae. The effect of epinephrine, however, does not seem to be mediated by ciliary muscle tone. We investigated the scleral spur in 37 human eyes (age 17-87 years). Serial tangential sections were studied with ultrastructural and immunocytochemical methods. The ciliary muscle cells do not enter the scleral spur, but their tendons, which consist of elastic fibers, are continuous with the elastic fibers in the scleral spur. Within the scleral spur is found a population of circularly oriented and spindle-shaped cells. The scleral spur cells form no bundles, but are loosely aggregated. They have long cytoplasmic processes and are connected to each other by adherence type and gap junctions. The scleral spur cells show intense staining for smooth-muscle-specific alpha-actin and myosin. Ultrastructurally, the scleral spur cells contain abundant actin filaments, but otherwise do not show the typical ultrastructure of ciliary muscle cells. The scleral spur cells do not express a complete basal lamina. They form individual tendinous connections with the elastic fibers in the scleral spur, which are continuous with the elastic fibers of the trabecular meshwork. The scleral spur cells are in close contact with nerve terminals containing small granular vesicles that are typical for adrenergic terminals. We conclude that the scleral spur cells are contractile myofibroblasts. Their contraction might be involved in the effects of epinephrine on the aqueous outflow.