The aim of this study is to examine the interrelationship of pituitary and ovarian hormone deficiency on the regulation of bone growth and bone formation rate. 48 female rats, at 3 months of age, were divided into age-matched intact control, hypophysectomized (HX), ovariectomized (OV), and HX + OV groups. Ten rats were killed at 3 months of age as baseline controls, and the rest of the animals were killed 5 weeks after surgery. Serum levels of osteocalcin and dynamic histomorphometry on the periosteal surface of the tibial shaft and fifth lumbar vertebrae were measured to evaluate systemic and local bone turnover. Tibial and fourth lumbar vertebral bone area, bone mineral content, and bone density were measured by dual-energy X-ray absorptiometry (DXA). Our results confirmed that OV increased and HX suppressed systemic and periosteal bone formation parameters in both bone sites, OV increased and HX suppressed the gain in bone size and bone mass. When OV rats were HX, the serum levels of osteocalcin and periosteal bone formation parameters of the tibial shaft and the fifth lumbar vertebrae were, however, depressed and did not differ from that of the HX alone. DXA results show that the effect of OV on bone size and bone mass is also abolished by HX. In conclusion, we have demonstrated that OV increases tibial and lumbar vertebral bone formation and bone growth and this effect is pituitary hormone dependent.