A Rapid Bioassay for Measuring Inhibin Activity

Abstract

Swiss albino female mice, 27 days old, weighing 25-30 g, were primed with 20 IU hCG [human chorionic gonadotropin], given in 2 equal doses, administered at 0900 h and 1800 h. An increment of 200% in uterine weight could be observed when the animals were autopsied the next day at 1000 h. The validity of the endpoint chosen, as a function dependent on FSH [follitropin], was shown by the fact that neutralization of endogenous FSH by FSH antiserum led to an inhibition of uterine weight increase. Further injection of the inhibin material caused an actual reduction in the the heightened levels of FSH (ng/ml) brought about by hCG: Control, 207 .+-. 13.6; hCG, 365 .+-. 28.8 and hCG + inhibin, below the detection level of radioimmunoassay. Inhibin preparation from ovine testicular extract when tested at 3 dose levels showed a dose dependent and significant suppression in uterine weight increase. The assay was statistically valid .epsilon. = 2.5; slope = 23.9; .lambda. = 0.104 and intra- and interassay variation = 8.3% and 11.7%, respectively. Since the mouse uterine weight assay is specific, has greater sensitivity and is uniformly reproducible, this assay procedure is ideally suited to assess the activity of different inhibin test preparations as well as to follow the purification of inhibin activity from crude material.