Quantitation of prolactin-dependent responses in porcine mammary explants

A system was developed to quantitate prolactin-dependent responses in porcine mammary tissue obtained from pregnant gilts. Metabolic responses to prolactin (Prl) and cortisol (C) in the presence of varying doses of insulin (I) were examined in mammary explants cultured on the surface of the medium or submerged in medium, under an atmosphere of humidified air. Explants suspended on grids at the surface of medium oxidized 45% more glucose (P less than .05) and incorporated 67% more glucose into lipids (P less than .05) than explants submerged in culture medium. In explants cultured on grids, both 100 and 1,000 ng I/ml increased glucose oxidation (by 50%) and glucose incorporation into lipids (by 150%) compared with 10 ng/ml (P less than .05), but responses to 100 and 1,000 ng I/ml were not different. Therefore, in subsequent studies, explants were cultured on grids with 100 ng I/ml. Rates of glucose metabolism for mammary explants cultured with I + C for 48 or 72 h were not different from those in fresh tissue. However, addition of Prl (200 or 1,000 ng/ml) increased oxidation rate 130% and fat synthesis 400% compared with I + C (P less than .05). Addition of triiodothyronine to I + C + Prl further increased rate of fat synthesis by 87%. Dose-dependent responses to Prl were demonstrated and were within the concentrations of Prl found in blood of gestating gilts. These studies demonstrate that the lactogenic complex of I, C and Prl induces metabolic activity in porcine mammary tissue from late pregnancy.