Diurnal variations in serum hormone levels during 2 different stages of prepubertal development were investigated in male and female rats. Groups of 13 to 18 and 25 to 30 day old male and female rats were decapitated at 4-hour by intervals during a period of 24 h. Their blood was collected and hormones were measured by radio-immunoassay. FSH levels were constantly high in 13 to 18, but low in 25 to 30 day old females. FSH was low in younger males, and significantly higher but without diurnal fluctuations in the older males. Serum LH was low in approximately 40% of the 13 to 18 day old females, while 40% had moderately high levels, and the remaining females extremely high levels of the hormone. Most of the extremely high LH peaks were found at 15.00 h and some at 03.00 h. Older females and males of both age groups had constantly low serum LH levels. Serum oestradiol was high in males and females during days 13 to 18, but it was lower in the 25 to 30 day old animals. In the young females prolactin was slightly elevated between 15.00 h and 19.00 h, while in the males the serum prolactin fluctuations were not significant. Serum testosterone was low in females at all times. The 13 to 18 day old males had higher testosterone levels than the 25 to 30 day old males. Both groups showed slight, but insignificant fluctuations in serum testosterone.