Electrophoretic separation of forms of rat growth hormone with different bioassay and radioimmunoassay activities: comparison of intraglandular and secreted forms

Abstract

Rat adenohypophysial tissue and medium in which pituitary explants had been incubated were processed by polyacrylamide disc gel electrophoresis (pH 9.5). Eluates of 6 regions of the gel columns were tested for growth hormone (GH) by a bioassay (BA; rat tibia test) and a radioimmunoassay (RIA). In the tissue, both assays detected most of the GH (80–99%) in segments with an R f range of 0.30–0.56. In the medium, both assays recorded a more cathodal distribution of most of the hormone (85–96%) in the R f region 0.00–0.42. The BA/RIA ratios of the segment containing the prominent stainable GH band (R f range 0.30–0.42) varied from 0.6–1.8 in tissue and medium. The tissue contained slow migrating (R f range 0–0.29) GH that had a high BA/RIA ratio (11.1–25.7). The medium contained high ratio GH (32.7–33.8) in a more anodal location (R f 0.57–0.68). Purified rat prolactin (RP-I) was not active in either the BA or the RIA. The results indicate that rat GH exists in electrophoretically heterogeneous forms which differ in their BA/RIA activity ratios. The proportion of these forms stored in adenohypophysial tissue and secreted by the gland may differ.