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The effects of constant light, constant darkness and diurnal lighting, in combination with pinealectomy or sham-pinealectomy, on pituitary and plasma concentrations of radioimmunoassayable prolactin were investigated in 8-week-old male and virgin female rats. Two to three days after operation random groups of pinealectomized and sham-pinealectomized animals of the same sex were placed together in either continous light, continuous darkness or diurnal light, and killed 21 days later. Compared with sham-operated diurnally-illuminated controls, constant darkness caused a decrease in pituitary prolactin content and a rise in plasma prolactin levels. Pinealectomy or constant illumination reversed the effect of constant darkness, resulting in an increase in pituitary prolactin content and a fall in plasma prolactin levels when compared with sham-operated diurnally-illuminated controls. Electron microscopy of lactotrophic cells of the sham-pinealectomized animals exposed to constant darkness revealed few cytoplasmic granules, whereas these cells in the sham-pinealectomized animals exposed to constant light contained abundant granules; compared with the former groups, lactotrophic cells of sham-pinealectomized rats exposed to diurnal lighting revealed an intermediate degree of granulation.