The effects of surgery and red light on prolactin (Prl) secretion were investigated in cycling and in pseudo-pregnant (PSP) rats. Secretion patterns of Prl were determined at hourly intervals from 07.00 to 22.00 h. Different regulatory mechanisms for Prl secretion were hypothesized for three time periods: a nocturnal (07.00–11.00 h), a prediurnal (14.00 -17.00 h), and a diurnal (19.00–22.00 h) period. The results demonstrate that red light can affect significantly Prl secretion, in particular nocturnal and prediurnal Prl secretion in estrous, diestrous day 1 and PSP rats. The effect of surgery varied with the time of the day and was dependent upon whether the animals, during the dark period, were maintained under full darkness or constant red dim illumination. In PSP rats the regulation of Prl secretion was different for the three time periods. In PSP day 0 rats there was a prediurnal surge of Prl secretion, comparable in timing and regulation to a prediurnal surge in estrous rats. This prediurnal surge was not evident on the other days of PSP. The regulation of nocturnal as well as diurnal Prl secretion was similar in PSP rats, but differed from cycling rats. The afternoon surge of Prl secretion on proestrus lasted the full afternoon and was basically one surge, distinct from all other surges. In diestrous rats Prl levels were low but showed a circadian variation. In summary, the effects of red light and surgery on Prl secretion varied with the physiological state and with the time of the day, indicating that the regulation of Prl secretion is complex and multimodal in nat