Sixteen prepubertal Holstein Friesian heifers were used to study the effect of long-term administration of bovine GH (bGH) on mammary development in intact and ovariectomized heifers. Eight heifers were ovariectomized at 2.5 months of age. Four intact and four ovariectomized heifers received subcutaneous injection of bGH (15 mg/day) for 15 weeks starting at 176 +/- 3 days of age (147 +/- 3 kg body weight), while the remaining eight heifers received an equal volume of excipient. Blood samples were collected weekly from 2 months of age. Heifers were slaughtered on the day after the last injection of bGH or excipient. Mammary gland development was quantified by dissection, chemical analysis and computer tomographic scanning. Mammary growth response at the time of slaughter was examined in cultures with explants prepared from parenchyma. Histological and histoautoradiographic studies with explants were performed. Treatment with bGH resulted in a significantly (P < 0.05) smaller mammary gland because of a reduced amount of extraparenchymal tissue. Ovariectomy markedly reduced the amount of parenchymal tissue. Growth response in mammary explants showed no treatment differences, suggesting that the decreased amount of parenchyma in ovariectomized heifers was caused by a decrease in mammary cell proliferation occurring some time prior to slaughter. The histological composition of mammary parenchyma was not changed by bGH treatment. However, ovariectomy resulted in less epithelial tissue (P < 0.001) and lumen (P < 0.05) and more stroma (P < 0.001), expressed as percentage tissue area. Serum hormone concentrations of bGH and insulin-like growth factor-I (IGF-I) were increased by bGH treatment in both intact and ovariectomized heifers.(ABSTRACT TRUNCATED AT 250 WORDS)