Effect of suckling intensity on human growth hormone binding, biochemical composition and histological characteristics of ovine mammary glands

Suckling both, or only one contralateral mammary gland during 15 days postpartum was utilized to study lactogenic hormone binding to mammary microsomal membranes and quantitative mammary morphology in ewes. Binding of radiolabeled human growth hormone was specific for lactogenic hormones. Non-radiolabeled human growth hormone, ovine and bovine prolactin and human placental lactogen effectively competed with radiolabeled human growth hormone for binding sites but ovine and bovine growth hormone were completely ineffective. Specific binding of radiolabeled human growth hormone to 600 μg of membrane protein averaged 23 ± 3% in all lactating glands. Neither days postpartum nor treatment of contralateral mammary glands substantially altered hormone binding in lactating glands. Specific human growth hormone binding (6 ± 0.5%) in non-suckled glands (15 days postpartum both udder halves) was significantly lower (P<0.01) than in lactating tissue but only a moderate and variable reduction in specific binding was measured in membranes from glands non-suckled for 15 days but contralateral to a suckled gland (14 ± 4%). Specific binding was approximately doubled in assays with 600 compared with 300 μg of membrane protein and the pattern of binding among variously suckled glands was not changed by treatment of membranes with 4 M MgCl₂ prior to assay. Most secretory cells from all lactating glands had rounded, basally displaced nuclei, apical fat globules, secretory vesicles and abundant densely stained basal cytoplasm (ergastoplasm). Alveolar lumenal area was maximal (50% of tissue area) and stromal tissue area was minimal. After 15 days of non-suckling (both udder halves) mammary cells were engorged with lipid, ergastoplasm was reduced and nuclei were irregularly shaped and randomly displaced compared with lactating tissue. In addition, lumenal area was reduced and stromal tissue more evident. Lack of suckling for 5 days had little apparent effect on mammary cytology. Like lactogenic hormone binding, mammary tissue morphology was only moderately altered by 15 days of non-suckling when the remaining gland was suckled. RNA concentration was lowest (2.1 ± 0.3 mg/g) in mammary tissue from ewes in which neither gland was suckled for 15 days postpartum but non-suckling interval had no significant effect when contralateral glands were suckled. DNA concentration was not significantly influenced by suckling treatments. Relative lactogenic hormone binding closely corresponded to changes in cytological and biochemical indices of secretory cell function.