Postnatal development of pituitary lactotropes in the rat measured by reverse hemolytic plaque assay

Developmental changes and sexual differences in the lactotrope population were studied with a reverse hemolytic plaque assay for detection and measurement of PRL secretion from individual cells in conjunction with PRL immunocytochemistry (ICC). Pituitary glands from both sexes at different ages were monodispersed with 0.1% trypsin. Freshly dispersed cells were incubated in Cunningham chambers for measurement of the fraction of plaque-forming cells and the size of plaques formed, or attached to glass slides for measurement of the fraction of cells staining for PRL by ICC. The percentage of plaque-forming cells in both sexes gradually increased with age from about 5% of the total anterior pituitary cell population at 5 days of age to the adult levels of about 54% in the females at proestrus and about 37% in the males. Sexual differences in the percentage of plaque-forming cells were consistently seen at 40 days of age and thereafter. The mean size of plaques in both sexes also increased with age from about 1,100 microns 2 at 5 days of age to a peak level of about 15,600 microns 2 in the females at proestrus and about 6,700 microns 2 in the males at 60 days of age; then fell to the adult levels of about 5,500 microns 2 in the females at proestrus and about 3,200 microns 2 in males. Sexual differences first appeared at 40 days of age and were estrogen-dependent. The results from ICC closely matched those from the plaque assay, except at 5 days of age and in the adult males. At 5 days, the fraction of cells staining for PRL was twice that of cells forming plaques. In adult male rats there were about 47% immunostained PRL cells but only about 39% plaque-forming cells. This difference, however, disappeared after estrogen treatment. These results demonstrate that sexual differences in the lactotrope population develop around puberty in rats and appear to be estrogen dependent.