The calcium homeostatic mechanism was challenged in adult female rats by feeding them a calcium-deficient diet containing oxalate, and by subjecting them to pregnancy and lactation. The regimen caused a substantial weight loss, especially in those animals which reared their young well. Severe hypocalcaemia was observed in the lactating rats. Serum-P was slightly elevated. The content of hydroxyproline in serum was considerably elevated, reflecting the degree of calcium deprivation. Serum proteins were least influenced. The calcium depriving regimen reduced the growth of long bones, but did not stop it. The ash content of the bones was considerably reduced, the degree of reduction depended on the degree of calcium deprivation. Ash as percentage of total bone organ was reduced, but not to the same extent as total ash. Analyses of different parts of femur showed that the proximal and distal parts had lost more bone mineral than the diaphyseal shaft. The ash content of cortical bone tissue from the femur was estimated by a volumetric method. No differences were observed between test groups and controls, indicating that no measurable amounts of bone mineral had been removed from the walls of the vascular canals or by osteocytic osteolysis. Planimetric determinations on cross sections from femora disclosed that a great amount of bone had been removed from the endosteal surface of the diaphysis, while the periosteal surface demonstrated reduced bone apposition.