Five groups of five lactating rabbits each were used. Milk yield was recorded from the 8th day of lactation onwards and on the 10th day of lactation the rabbits received the following treatments: Group S, sham-operation with saline (1 ml/12 h); Group P, hypophysectomy with sheep prolactin (1 mg/12 h); Group H, hypophysectomy with human growth hormone (1 mg/12 h); Group B, hypophysectomy with bovine growth hormone (1 mg/12 h) and Group C, hypophysectomy with saline (1 ml/12 h). The injections of saline or hormones were continued for 5 days and at the end of this period a blood sample was taken, the animals were killed and their mammary glands removed for histological examination and assay of the following enzymes: 6-phosphogluconate dehydrogenase (EC 18.104.22.168), phosphofructokinase (EC 22.214.171.124), phosphoglucomutase (EC 126.96.36.199), UDP-glucose pyrophosphorylase (EC 188.8.131.52), acetyl-CoA carboxylase (EC 184.108.40.206), acetyl-CoA synthetase (EC 220.127.116.11) and ATP-citrate lyase (EC 18.104.22.168). On the 5th day after surgery the concentrations of blood l-lactate and pyruvate and plasma free fatty acids and protein were similar in all groups, whereas plasma glucose was higher in groups S, B and H than in groups P and C. Although the weights of pituitary target organs (adrenals, thyroid and ovaries) were similar in all groups, the weights (g/kg body weight) of mammary tissue varied markedly, group S being the heaviest and group C the lightest. Milk yields, 5 days after surgery, for groups P and H were about 50% that for S, whereas those for B and C were 15 and 4% respectively. Where possible the enzyme activity was expressed as a ratio of the rate of synthesis of the end product of the pathway in which the enzyme occurred. With the exception of acetyl-CoA carboxylase which may have had a rate-limiting role in the synthesis of milk fat, enzymic activity in vitro was in excess of that required in vivo for the synthesis of either milk fat or lactose. It appeared that the rate of milk synthesis depended upon the degree of maintenance of the secretory epithelial cells within the mammary gland rather than a block in the synthetic pathways within these cells.