The production of shell-less eggs was induced in hens to measure the effects of the high demands made by shell formation on the blood minerals and hormones whose concentrations change during egg formation. In control hens laying hard-shelled eggs, the concentration of ionized calcium in plasma decreased at the onset of shell formation, but no change was found in hens laying shell-less eggs. Total calcium concentrations in plasma decreased slightly throughout the ovulation cycle in both groups. Concentrations of inorganic phosphorus in the plasma were increased in the control group during the period of shell formation and decreased when calcification was suppressed. Finally, the concentrations of 1,25-dihydroxycholecalciferol (1,25-(OH)2D3) in plasma were significantly increased 16 and 20 h following an ovulation compared with 4 h after ovulation, or compared with the concentrations observed in hens laying shell-less eggs. The variations in the plasma concentrations of ionized calcium, inorganic phosphorus and 1,25-(OH)2D3 associated with egg formation were therefore absent in hens laying shell-less eggs demonstrating their direct link with shell calcification. On the other hand, suppression of shell production had no influence on the changes in the plasma concentrations of progesterone, oestradiol and testosterone which are associated with the normal ovulatory cycle. It is concluded that the increases in intestinal and uterine calcium transport and in 1,25-(OH)2D3 production which occur at the onset of egg production in hens are mainly controlled by factors involved in maintaining calcium homeostasis rather than by gonadal hormones.