Amplitude regulation of episodic release, in vitro biological to immunological ratio, and median charge of human chorionic gonadotropin in pregnancy

In the present study, we examined the regulation of 24-h serum immunoreactive levels, in vitro biological to immunological (B/I) ratio, and median charge of circulating CG at the end of the first, second, and third trimesters of human gestation. Seven pregnant women were prospectively studied at 12-15, 23-26, and 35-38 weeks of gestation. Blood was sampled every 20 min over a 24-h period, and serum CG concentrations were determined by RIA. Pulse detection and analysis of the 24-h rhythm of serum immunoreactive CG concentrations were carried out by the program Cluster and cosine curve fitting, respectively. The in vitro biological activity of circulating CG was determined by the mouse Leydig cell-testosterone production bioassay, and the median charge of its isoforms was determined by zone electrophoresis in agarose suspension. The immunoreactive levels of CG present at the end of each trimester of gestation fluctuated over a 24-h period; such variability exceeded that of the within-assay coefficient of variation of the CG RIA and could be resolved into a series of CG peaks and valleys. Although no trend in the number of peaks or valleys was systematically found in relation to gestational age, comparisons between the amplitude and area of the CG peaks revealed that these pulse parameters were significantly higher at 12-15 weeks than at 23-26 and 35-38 weeks of gestation. Cosine fits for 24-h rhythms revealed the existence of significant nyctohemeral profiles of serum CG levels in all women studied at 12-15 weeks, in four subjects at 23-26 weeks, and in six women at 35-38 weeks gestation. The time of acrophase was highly homogeneous only between 12-15 weeks of gestation, occurring between 1057-1452 h in six of the women. The in vitro B/I ratio of CG contained in serum pools from 12-15 weeks was significantly (P < 0.05) higher than that exhibited by CG during later gestational periods (B/I ratio at the end of first trimester, 1.14 ± 0.14; second trimester, 0.87 ± 0.22; third trimester, 0.79 ± 0.12). hCG isoforms at 12-15 weeks were more negatively charged than those circulating at 23-26 and 35-38 weeks of gestation. There were no significant differences between the B/I ratio and the median charge of CG molecules from the second and third trimesters. We conclude that serial serum concentrations of CG throughout pregnancy show significant amplitude-modulated pulsatile release and nyctohemeral variations. The amplitude-regulated pulsatile mode of release is probably endowed by the placenta. These changes in episodic CG release throughout gestation are concomitantly accompanied by alterations in the molecular structure of the gonadotropin, thus indicating the existence of a variety of distinct mechanisms through which the placenta regulates the intensity, biochemical composition, and/or clearance of the CG signal.