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It is well known that gonadotropin controls a major part of follicular development. However, the mechanism of local regulation under the control of gonadotropins in still unclear. In this study, we focused on the local regulation of steroidogenesis, growth factors and cell proliferation to evaluate the human follicular development. To assess steroidogenesis, it is important to detect the expression of steroidogenic enzymes in the granulosa and theca cells during folliculogenesis. We initially tried to find out the transcription factor Ad4BP that binds the Ad4 site and regulates the function of steroidogenic enzyme. By immunohistochemistry, the expression of Ad4BP was confirmed sporaf1p4lly in preantral granulosa cells. In the antral follicles, the expression of Ad4BP was observed both in the granulosa and theca cell. According steroidogenic enzyme, we evaluated temporal and spatial localization of cholesterol side chain cleavage (scc), 3 beta hydroxysteroid dehydrogenase (3 beta HSD), 17 alpha hydroxylase (17 alpha) and aromatase, and steroid receptors. In briefly, the localizations of scc, 3 beta HSD and 17 alpha were observed in preantral follicles and the mRNA expressions of these enzymes were confirmed in the theca cell by in situ hybridization method. Expression of aromatase was generally observed in only one follicle (antral or mature follicle) per case in mid proliferative to premenstrual phase. The localization of androgen and estrogen receptor was observed in the antral follicle granulosa cells, and estrogen receptor was detected only in aromatase positive follicles. These results suggested that Ad4BP initially controls the function of steroidogenic enzymes and steroidogenic enzymes gradually express from primary follicles to mature follicles. At antral follicle stage, steroid metabolism completes to produce testosterone. When aromatase and estrogen receptor express in antral follicle, this antral follicle develops as the dominant follicle and produces estradiol to promote follicle maturation. We therefore speculate that the expression of aromatase and estrogen receptor have an important role for the selection of dominant follicle in human. According growth factors for follicular development, it has been demonstrated to be important in the biological activity in the ovary. In this study, we examined the localization of EGF, TGF alpha and their receptor (EGFR). The localization of EGF was not confirmed both mRNA and protein level through follicular development. On the other hand, the localization and expression of TGF alpha was confirmed in theca cells and EGFR in granulosa cells at antral stage.(ABSTRACT TRUNCATED AT 400 WORDS)

We measured FSH and LH concentrations in 130 cord sera and 217 peripheral sera obtained from infants of 28-42 weeks in gestation, aged 5-75 days after birth, by radioimmunoassay. Cord FSH and LH levels were 3.9-13.6 mIU/ml and 43.3-88.5 mIU/ml respectively, and decreased with advancing gestation. Although FSH levels in 28-31 weeks were higher in female than in male, another sex difference was not found. Postnatal FSH levels in male preterm infants decreased within 10 day, thereafter continued low levels (3.7-5.6 mIU/ml). However, in male term infants and female infants, FSH levels increased with peak levels between 11-30 days after birth, and then decreased gradually. The elevation of FSH levels in female was more remarked and prolonged in preterm infants than in term infants. Postnatal LH levels in both male and female decreased rapidly after birth, and then had similar pattern of FSH levels. Serum gonadotropin levels during the neonatal period were higher in female than in male, except for LH in term infants (male greater than female).

Serum samples obtained from 18 normal men and women, 24 pregnant women of the third trimester, and 8 patients with galactorrhea-amenorrhea syndrome (GAS) were subjected to a 3-phase methanol extraction. The serum extract released prolactin (PRL) specifically from rat anterior pituitary in vitro. The PRL releasing activity (PRA) was dependent on the incubation time and dose of serum extract, while independent on TRH or PRL in serum. The mean PRA in serum of pregnant women was significantly (p less than 0.02) higher than one of normal subjects. The mean PRA in serum of GAS was significantly higher than in pregnant women (p less than 0.05), and in normal subjects (p less than 0.001). When the individual PRL levels were compared to the PRA in samples from pregnant women and normal subjects, a significant (p less than 0.02) positive correlation evolved. These results indicate that a PRL releasing substance(s) distinct from TRH is (are) present in serum, and has (have) a relationship to the PRL concentration and late pregnant states.

We measured maternal venous (MV), umbilical arterial (UA) and umbilical venous (UV) cortisol levels of 180 pregnancies at term, and investigated especially cases of fetal distress and neonatal asphyxia. No fetal distress and no neonatal asphyxia cases followed spontaneous vaginal deliveries (94), cesarean sections (18) or breech extractions (6). In cesarean section delivery, the cortisol levels of three individual samples were lower than in other modes of deliveries. The cortisol level of the umbilical artery was significantly higher than that of the umbilical vein in spontaneous vaginal delivery and cesarean section delivery. When spontaneous vaginal deliveries were classified into the fetal distress group and no fetal distress group, the cortisol levels of the umbilical artery and umbilical vein in the former group were significantly higher than those in the latter group. Furthermore, regardless of fetal distress there were significant positive correlations between the cortisol levels of MV & UA, MV & UV, and UA & UV. In addition, as the cortisol level of the maternal vein increased, the umbilical arterial-venous cortisol ratio (UA/UV) gradually approached 1.0. These data suggest that in fetal distress or neonatal asphyxia, the fetus may produce cortisol for itself independent of the mother.

We measured maternal venous(MV), umbilical arterial(UA), and umbilical venous(UV) cortisol levels of 180 pregnancies at term. In vacuum extraction cases, the cortisol levels of three individual samples were significantly higher than in other modes of delivery. In cesarean section delivery the cortisol levels of three individual samples were the lowest of all modes of delivery. The cortisol level of MV was higher than that of UA and UV in all deliveries. The cortisol levels of three samples showed no difference between the induced labor group and the spontaneous onset of labor group regardless of the delivery mode. In spontaneous vaginal delivery the cortisol levels of three individual samples were significantly higher in primipara than in multipara. Positive correlation was observed in all modes of delivery between the cortisol levels of MV & UA, MV & UV, and UA & UV. There was a negative correlation between the cortisol levels of MV and umbilical arteriovenous cortisol difference in multipara spontaneous vaginal deliveries. There was also a negative correlation between cortisol levels of MV and the umbilical arterial/venous ratio. These data suggest that the fetus may produce cortisol by itself and maternal cortisol may inflow to the fetus.

Adrenocorticotropic hormone (ACTH), beta-lipotropin (beta-LPH) immunoreactivity (IR) and beta-endorphin (beta-EP) immunoreactivity (IR) were measured by high-sensitive radioimmunoassay in maternal and umbilical cord plasma samples which were obtained simultaneously in 12 cases. Mean IR-beta-LPH in maternal (690.1 +/- 138.6 pg/ml, +/- S.E.) and cord venous plasma (1114.8 +/- 94.2 pg/ml) were significantly higher than that of normal adults (93.0 +/- 8.2 pg/ml). Mean IR-beta -EP in maternal (125.8 +/- 24.0 pg/ml) and cord venous plasma (130.6 +/- 20.6 pg/ml) were significantly higher than that of normal adults (6.0 +/- 0.9 pg/ml). There were significant positive correlations between IR-beta-LPH and IR-beta-EP in maternal and cord venous plasma. Mean beta-EP to beta-LPH molar ratio of 0.19 +/- 0.03 in maternal plasma was significantly (p less than 0.05) higher than that of cord venous plasma (0.13 +/- 0.01). There was no significant correlation between the levels of these three hormones in maternal plasma and the levels in cord venous plasma. These data suggest that ACTH, IR-beta-LPH and IR-beta-EP were elevate during labor, delivery responding the stress. Beta-LPH and beta-EP in cord venous plasma were fetal and/or placental origin.

Cord venous growth hormone (GH) and somatostatin levels were measured by radioimmunoassay in 86 infants and 11 anencephalic infants. 1) Cord GH levels were 69.2 +/- 20.2 ng/ml (N = 6, gestational weeks (GW) : 21-30W, mean +/- S.D.), 31.5 +/- 24.3 ng/ml (N = 11, GW : 31-36W), and 17.6 +/- 8.9 ng/ml (N = 71, GW: 37-41). In anencephalic infants, cord GH levels were significantly lower (2.3 +/- 1.4 ng/ml) than those in preterm and fullterm infants. 2) In fullterm infants, cord GH levels of small infants (19.2 +/- 9.2 ng/ml, N = 33, birth weight less than 3,250g) were significantly higher than that of large infants (15.1 +/- 8.1 ng/ml, N = 38, birth weight greater than of equal too 3,250g). On the other hands, cord somatostatin levels were relatively lower in small infants than in large infants. 3) There was a significant difference between male cord GH levels (20.6 +/- 8.9 ng/ml, N = 35) and female cord GH levels (14.6 +/- 8.0 ng/ml, N = 36). These results suggest that fetal GH secretion is suppressed by GH inhibiting factor (somatotropin release inhibiting factor) from mid gestation toward term and there is a sex difference of fetal GH secretion in fullterm fetus.

Prolactin (PRL), growth hormone (GH), and estradiol-17 beta (E2-17 beta) levels were measured by radioimmunoassay in umbilical venous serum from 118 full term normal infants and 11 anencephalic infants. In anencephalic infants, the concentration of GH:2.4 +/- 1.6 ng/ml (mean +/- SD, n = 11) and E2-17 beta:2.3 +/- 1.4 ng/ml (n = 6) were significantly lower than those in normal infants (GH: 16.5 +/- 9.0 ng/nl n = 65; E2-17 beta:12.4 +/- 8.0 ng/ml, n = 108). There was no significant difference of PRL levels between normal infants (324 +/- 118 ng/ml, n = 118) and anencephalic infants (253 +/- 86 ng/ml, n = 10). There was significant positive correlations between PRL and E2-17 beta levels in normal infants (p less than 0.01), and was significant negative correlation between PRL levels and birth weight (p less than 0.01). These results suggested that PRL secretion is affected by estrogens during fetal period and fetal PRL has some effects for the regulation of fetal growth as somatogenic hormone.

We measured maternal venous unconjugated estriol (UE3) levels in 244 cases (218 cases with labor and 26 without labor). We investigated the relationships between several obstetrical factors and UE3 levels. There was no significant difference between the group with labor and the group without labor. However, in multipara, the UE3 level fell gradually with prolonged duration of labor. There were significant positive correlations between the UE3 levels and birth weights (n = 82, r = 0.375, p less than 0.001), and placental weights (n = 82, r = 0.381, p less than 0.001) in multipara with labor. There was no significant difference according to sex between the UE3 levels of a mother carrying a male or female fetus. The UE3 levels of fetal distress cases (n = 30, mean +/- S.E. 20.0 +/- 8.6 ng/ml) were significantly (p less than 0.02) lower than those without fetal distress (n = 188, 24.5 +/- 9.7 ng/ml). These data suggest that after the onset of labor, maternal venous UE3 has some relationship to feto-placental function.

Serum testosterone (T) concentrations in 106 umbilical venous bloods and 104 neonatal venous bloods, which were obtained from preterm (32-36w) and term (36-41w) infants, were measured by RIA. Cord T levels of boys were higher in preterm infants than in term infants, and those of preterm infants were higher in boys than in girls. Neonatal T levels of girls decreased following delivery. But those of boys increased after 10 days of life. T levels were higher in boys than in girls at 11-30 days of life. Differences between preterm and term infants were not significant.

The changes in serum specific hCG and prolactin levels were studied in 20 women after the evacuation of normal pregnancy at 5-10 weeks (group A) and 10 patients after the evacuation of molar pregnancy of 9-16 weeks (group B). The elimination of circulating hCG was faster in group A than in group B, and that in women after term delivery(group C)was faster than in both groups A and B. The half-life of the serum hCG level was 44.4 hrs in group A, 42.7hrs in group B, and 38.9hrs in group C, respectively. The mean level of serum prolactin in group A was 27.4ng/ml at 0 day, 64.5ng/ml at 4-7 days, and 31.9ng/ml at 13-15 days after evacuation, respectively. The surge was statistically significant. Though no regular tendency was found in the prolactin levels in group B, the levels were relatively higher than in group A and nonpregnant women. No correlation was found between hCG and prolactin levels in either group A or group B. The administration of drugs, such as methotrexate, did not affect the prolactin levels. On BBT, the resumption of ovulation tended to occur when the prolactin level, rather than the hCG level, returned to its normal range.

Human granulosa-luteal cells obtained from dominant preovulatory follicles at the time of oocyte retrieval for in vitro fertilization (about 10 h after the peak of the endogenous LH surge) produced large quantities of progesterone (P) (8.7 pg/cell/first 2 days) in a monolayer culture. Although basal production of estradiol (E2) in these granulosa-luteal cells was restricted to small quantities, a marked increase in E2 production occurred in the presence of testosterone (T) (10(-6) M) as aromatizable substrate. The non-aromatizable androgen, 5 alpha-dihydrotestosterone (DHT) (10(-7) M, 10(-6) M) slightly enhanced E2 production in these granulosa-luteal cells and it did not inhibit T aromatization. In contrast, DHT did not increase E2 production in cultured granulosa cells obtained from follicles 2 approximately 5 days before the LH surge. Thus, the response of granulosa cells to exogenous androgen seems to vary at different stages of cell differentiation. Neither T nor DHT elicited significant changes in the production of P by granulosa-luteal cells obtained from preovulatory follicles during the LH surge. However, T (10(-6) M) suppressed hCG-stimulated P production in these cells. The results suggest that androgen enhances the estrogen biosynthesis of granulosa-luteal cells obtained from preovulatory follicles during the LH surge not only by acting as substrate for aromatization but also by participating in some process of estrogen synthesis. Further, it is suggested that androgen has a negative effect on P biosynthesis in these cells.

Lutenizing hormone (LH-hCG), follicle stimulating hormone (FSH), beta-human chorionic gonadotropin (beta-hCG), estradiol, cortisol and testosterone were determined and correlated with each hormones in 62 cord sera (32 male and 35 female infants). Mean (+/- S.E.)male cord sera cortisol concentrations (36.2 +/- 3.9 ug/dl) were significantly (p less than 0.05) higher than that of female (24.1 +/- 3.7 ug/dl). Another hormones were not found significant sex difference. Regression analysis showed significant positive correlations between LH-hCG (r = 0.525, p less than 0.001) or beta-hCG (r = 0.461, p less than 0.005) levels and FSH levels in all cord sera (N = 67). In male cord sera, there were significant positive correlations between LH-hCG (r = 0.493, p less than 0.01) or beta-hCG (r = 0.485, p less than 0.01) levels and testosterone levels. There was a significant positive correlation (p less than 0.01) between estradiol levels and testosterone levels in female cord sera. These data suggest (1) the sex difference of cortisol levels which indicate the response for the stress during labor and delivery, (2) there was a significant sex difference in the maturation of the feed back mechanism in pituitary-gonadal axis.