High physiological prolactin (PRL) stimulated intestinal calcium absorption and renal calcium uptake in mammals. Previous histomorphometric study revealed a significant increase in bone turnover in the trabecular part of the PRL-exposed long (cortical) bone; however, whole-bone densitometric analysis was unable to demonstrate such effect. We therefore studied differential changes in bone mineral density (BMD) and contents (BMC) of the femoral diaphysis and metaphysis in adult female rats exposed to high PRL induced by anterior pituitary (AP) transplantation. The estrogen-dependent effects of PRL on the femur were also investigated. We found that chronic exposure to PRL had no effect on BMD or BMC of the femoral diaphysis, which represented the cortical part of the long bone. It is interesting that 7 weeks after an AP transplantation, BMD and BMC of the femoral metaphysis were significantly decreased by 8% and 14%, respectively. Ovariectomy (Ovx) for 2, 5, and 7 weeks also decreased BMD and BMC in the femoral metaphysis, but not in the diaphysis. However, the AP transplantation plus Ovx (AP+Ovx) produced no additive effects. Nevertheless, 2.5 µg/kg 17β-estradiol (E2) supplementation abolished the osteopenic effects of both Ovx and AP+Ovx on the femur. As for the L5-6 vertebrae, BMD and BMC were not affected by PRL exposure, but were significantly decreased by Ovx and AP+Ovx, and such decreases were completely prevented by E2 supplementation. It could be concluded that high physiological PRL induced a significant osteopenia in the trabecular part, i.e., the metaphysis, of the femora of adult female rats in an estrogen-dependent manner. Since PRL had no detectable effect on the vertebrae, the effects of PRL on bone appeared to be site-specific.Key words: diaphysis, femur, metaphysis, prolactin, vertebrae.Prolactin (PRL), as a calcium-regulating hormone, has been reported to stimulate intestinal calcium absorption, enhance renal calcium reabsorption, and induce high bone turnover in nonmated female rats [1][2][3]. By regulating calcium mobilization in these target organs during pregnancy and lactation, the high physiological PRL of 75-300 ng/ ml increases calcium availability for fetal growth and milk production [4]. Moreover, the basal PRL level of 7-10 ng/ ml was required for the maintenance of normal bone turnover and duodenal calcium absorption in rats [5,6].It has been shown that PRL acted directly on the intestinal absorptive cells to enhance calcium absorption [3]. In bone, osteoblasts but not osteoclasts strongly expressed functional PRL receptors (PRLR), indicating that bone was also a target of PRL [7]. However, the direct effect of PRL on osteoblasts to enhance bone turnover was complicated by its indirect actions through hyperprolactinemiainduced hypogonadism, which in turn induced chronic estrogen deficiency [8,9]. Moreover, the PRL-enhanced calcium absorption may indirectly contribute to the increase in bone calcium deposition, as previously suggested by the 45 Ca kinetic and histomorphometri...