Our knowledge of the developmental changes in the concentration of serum and bone osteocalcin (OC) is limited. To investigate the interrelationship between skeletal and circulatory OC during acquisition of peak bone density in mice, we examined the temporal changes in the concentration of serum and bone OC from 3 to 12 weeks of age between C3H/HeJ (C3H) and C57BL/6J (B6), two commonly used inbred strains of mice with a large difference in bone density. We have demonstrated an increase in bone and decrease in serum OC during the acquisition of peak bone density in C3H and B6 mice which parallels an increase in bone mineral density. These two strains exhibited differential changes in the concentration of OC. C3H mice retained more OC in bone and secreted less into serum compared with B6, which coincides with the large differences in bone density between these two strains. These opposite changes of OC levels in bone and serum between C3H and B6 stress the importance of defining the genetic mechanisms underlying the differences in OC metabolism, differences that could be relevant to the acquisition and maintenance of bone mass in mice.