In humans and other primates, an increase in luminance in either eye elicits bilateral pupilloconstriction that is essentially equal in both eyes. Current models of the neural substrate for this clinically important light reflex propose that a retinorecipient pretectal nucleus projects bilaterally to the Edinger-Westphal nucleus (EW), which contains the parasympathetic, preganglionic neurons controlling pupilloconstriction. Based on single-unit recording studies in anesthetized cats and rats, it has been further suggested that luminance neurons in only one pretectal nucleus, the pretectal olivary nucleus, mediate this reflex. However, to our knowledge, there have been no comparable electrophysiological studies in primates of the pupillary light reflex or the pretectal luminance neurons that mediate this reflex. To address this issue, single-unit recording and electrical microstimulation studies were carried out in the pretectum of alert, trained, rhesus monkeys. These studies demonstrated that the primate pretectum contains luminance neurons with the characteristics appropriate for mediating the pupillary light reflex and that these neurons are located in one retinorecipient pretectal nucleus, the pretectal olivary nucleus. Electrical microstimulation at the site of these neurons often elicited pupilloconstriction. Our results provide clear evidence for the involvement of the pretectum, and more specifically the pretectal olivary nucleus, in mediating the pupillary light reflex in primates. © 1995 Springer-Verlag.