Since endogenous prolactin has been shown to enhance food consumption, calcium absorption, and bone calcium turnover in the pregnant rat, the role of endogenous prolactin in the regulation of calcium metabolism was investigated in 3-day balance studies of female Wistar rats from the age of 3 to 11 weeks. The study was divided into two parts. In part I, calcium metabolism in males and females was compared. In part II, 3-week old female rats were divided into 5 groups: (i) control animals receiving 0.9% NaCl; (ii) animals receiving 6 mg bromocriptine/kg/day (- PRLendo group); (iii) animals receiving 2.5 mg ovine prolactin/kg/day (+PRLexo); (iv) sham-operated animals receiving 0.9% NaCl, and (v) animals with two extra pituitaries implanted under the renal capsule, receiving 0.9% NaCl (AP group). Results showed that rapid growth occurred between 3 and 6 weeks with maximum fractional calcium absorption and calcium retention at 5 weeks of age in both sexes. The data also showed a physiological significance of endogenous prolactin in enhancing calcium absorption and retention in 5 week old rats. In an absence of prolactin, peak calcium absorption was delayed in 7-week old animals, and vertebral calcium content of 11-week old animals was reduced by 18%. Hyperprolactinemia in the AP group was found to enhance fractional calcium absorption and calcium retention at 7, 9, and 11 weeks and increased the femoral calcium content by 16%. It could be concluded that a physiological role of prolactin is the stimulation of calcium absorption and maintainance of bone calcium content during growth and development.