The biogenic amine octopamine (OA) modulates invertebrate behavior by changing neuronal responses from sensory inputs to motor outputs. However, the OA modulation of visual sensitivity and its possible coupling to diurnal cycles remains unexplored. Here we studied the diurnal variations in the OA levels in the hemolymph of the crayfish Procambarus clarkii, its release from the structures in the eyestalk and its modulation of the retinal light sensitivity. The hemolymph concentration of OA and its amino acid precursor tyrosine was measured by high-resolution liquid chromatography; OA varied along the 24-h cycle. The peak value appeared about 2 h before the light offset which preceded the peak locomotor activity. OA was found in every structure of the eyestalk but displayed higher levels in the retina–lamina ganglionaris. Moreover, OA was released from isolated eyestalks at a rate of 92 nmol/eyestalk/min and a calcium-dependent release was evoked by incubation in a high potassium solution. OA injected into dark-adapted crayfish or applied to the isolated retina at concentrations of 1, 10 and 100 μM produced a proportionally increasing reduction in the amplitude of the photoreceptor light responses. These OA concentrations did not affect the position of the visual accessory pigments. Our results suggest that OA release in the crayfish eyestalk is coupled to the 24-h cycle to regulate the diurnal reduction of the photoreceptor sensitivity and to favor the expression of exploratory locomotion during the dark phase of the circadian cycle.