Immunoreactive corticotropin-releasing hormone in human plasma during pregnancy, labor, and delivery

We previously reported that immunoreactive corticotropin-releasing hormone (CRH) is present in human placenta and third trimester maternal plasma, and that such material is very similar to rat CRH and the predicted structure of human CRH. We suggested that maternal plasma immunoreactive CRH may be of placental origin. To further investigate this possibility, we measured plasma immunoreactive CRH in women during pregnancy, labor, and delivery and 1 and 2 h postpartum, and in nonpregnant women. Umbilical cord plasma and placental CRH concentrations were also measured. In the first trimester of pregnancy, the mean maternal plasma level was 5.9 +/- 1.0 pg (+/- SEM)/ml (n = 24), not significantly different from that in 10 nonpregnant women (5.8 +/- 0.8 pg/ml). Plasma CRH concentrations progressively increased during pregnancy (second trimester, 35.4 +/- 5.9 pg/ml (n = 39); early third trimester (28-34 weeks), 263 +/- 41 pg/ml (n = 14); late third trimester (35-40 weeks), 800 +/- 163 pg/ml (n = 20)]. There was a significant correlation between maternal plasma CRH levels and weeks of pregnancy. Plasma CRH concentrations were further elevated (2215 +/- 329 pg/ml; n = 9). During early labor, peaked at delivery (4409 +/- 591 pg/ml; n = 28), and declined rapidly after delivery [1 h postpartum, 1042 +/- (353 pg/ml (n = 13); 2 h postpartum, 346 +/- 91 pg/ml (n = 13)]. There was a significant correlation (r = 0.562; P less than 0.01) between matched maternal plasma and placental CRH concentrations. The mean umbilical cord plasma CRH level (50.6 +/- 6.1 pg/ml; n = 28) was much lower than that in the mother at the time of delivery. Umbilical venous plasma CRH levels were significantly greater than those in simultaneously obtained umbilical arterial plasma (70.8 +/- 11.3 and 41.8 +/- 4.9 pg/ml, respectively; n = 11). There was a significant correlation (r = 0.384; P less than 0.05) between maternal and fetal CRH concentrations. Gel filtration of plasma obtained from women during the third trimester, at delivery, and early postpartum and placental extracts revealed two major peaks of immunoreactive CRH: a high mol wt peak and one at the elution position of rat CRH. In contrast, only rat CRH-sized material was detected in plasma from nonpregnant women and umbilical cord plasma. Maternal plasma immunoreactive CRH-sized material stimulated ACTH release from anterior pituitary tissue in a dose-dependent manner and was equipotent with rat CRH.(ABSTRACT TRUNCATED AT 400 WORDS)     

Placental corticotropin-releasing hormone and pituitary-adrenal function during pregnancy.

The placenta secretes large amounts of the hypothalamic releasing hormone, corticotropin-releasing hormone (CRH), into both the maternal and fetal circulation during pregnancy. We characterized the relationship between maternal plasma CRH and products of the pituitary and adrenal in order to investigate the physiologic role of placental CRH in modulating maternal pituitary-adrenal function. Plasma was obtained from 8 women at biweekly intervals between 21 and 40 weeks of full-term pregnancy for CRH, adrenocorticotropin (ACTH), alpha-melanocyte-stimulating hormone (alpha MSH), cortisol, and dehydroepiandrosterone sulfate (DHEAS) measurements by radioimmunoassay. Eighteen women were also studied once at 22-34 weeks of pregnancy with plasma CRH and 24-hour urinary free cortisol measurement. Eight nonpregnant women served as control subjects. Plasma CRH was undetectable in the nonpregnant subjects and rose over the time period studied in the pregnant women. Concentrations of afternoon ACTH and cortisol also rose during pregnancy while DHEAS levels declined in the pregnant women. The alpha-MSH levels were beneath the level of detection (< 20 pg/ml) in both the pregnant and nonpregnant subjects. The overall mean afternoon ACTH concentration was higher in the pregnant than in the nonpregnant women (11.4 +/- 1.8 vs. 5.9 +/- 1.8 pg/ml; p < 0.05), although the ACTH levels in both groups remained within the normal range. The mean plasma cortisol concentrations were higher in the pregnant women, while the mean DHEAS levels were lower in the pregnant women when compared to the nonpregnant subjects.(ABSTRACT TRUNCATED AT 250 WORDS)     

Plasma corticotropin-releasing hormone concentrations during pregnancy and parturition

Plasma CRH was measured in maternal plasma throughout the third trimester of pregnancy, during labor, and postpartum. CRH levels were also measured in arterial and venous umbilical cord plasma samples. In normal pregnant women, plasma CRH increased from 50 +/- 15 (+/- SEM) pg/mL at 28 weeks gestation (n = 41) to 1462 +/- 182 pg/mL at 40 weeks (n = 55) and 1680 +/- 101 pg/mL (n = 65) in labor. Women with pregnancy-induced hypertension (n = 49) had plasma CRH levels significantly elevated above this normal range. Similarly, women who subsequently went into premature labor had raised levels several weeks before the onset of labor. After delivery, plasma CRH returned to normal within 15 h. Total plasma cortisol levels varied little throughout the third trimester, but increased during labor and remained elevated 2-3 days postpartum. There was, therefore, no correlation between plasma cortisol and CRH, implying that this placental CRH is not primarily involved in the control of the maternal hypothalamo-pituitary adrenal axis during pregnancy. The concentrations of CRH in umbilical cord plasma samples were considerably lower than those in the maternal circulation and were close to those in normal nonpregnant adults.     

Response to corticotropin-releasing hormone during pregnancy in the baboon

CRH is secreted by the placenta into human maternal and fetal plasma during gestation. In the present study plasma CRH was measured in the plasma of five pregnant baboons and their fetuses to ascertain whether the baboon is a suitable model for study of placental CRH. Studies were performed in chronically catheterized animals that exhibited no behavioral or endocrinological signs of stress; maternal animals moved freely about the cage. Mean maternal plasma CRH was 620 +/- 110 pmol/L (2970 pg/mL) at 146 +/- 11 days gestation, and mean fetal plasma CRH was 133 +/- 29 pmol/L (640 pg/mL) at delivery in four animals. Plasma CRH was undetectable (less than 8.5 pmol/L; less than 41 pg/mL) in nonpregnant animals and in animals 8 h after delivery. Maternal and fetal plasma CRH levels in the chronically catheterized baboon were very similar to human maternal and umbilical cord CRH levels at comparable gestational ages. In addition, the majority of maternal plasma CRH eluted in the same position as synthetic human CRH by gel filtration. CRH stimulation tests were performed in the chronically catheterized maternal baboon to investigate whether pituitary-adrenal function during pregnancy is similar to that observed after chronic CRH infusion; blunted ACTH and cortisol responses to acute injections of CRH are observed after chronic CRH infusion. The administration of 0.5 micrograms/kg ovine CRH (oCRH) failed to result in an ACTH or cortisol rise in four pregnant baboons. Baseline ACTH levels were 5.2 +/- 0.4 pmol/L (23.5 pg/mL), and baseline cortisol levels were 800 +/- 55 nmol/L (29.1 micrograms/dL); neither rose after CRH administration. In contrast, 0.5 micrograms/kg oCRH did result in significant ACTH and cortisol elevations in five nonpregnant baboons [ACTH: baseline, 5.9 +/- 1.4; peak, 16 +/- 4.8 pmol/L (P less than 0.05); cortisol: baseline, 430 +/- 55 nmol/L; peak, 960 +/- 200 nmol/L (P less than 0.05)]. In contrast, the administration of a larger dose of oCRH (5.0 micrograms/kg) led to stimulation of ACTH release in five pregnant baboons (baseline, 6.6 +/- 1.3 pmol/L; peak, 34.1 +/- 6.4; P less than 0.001). After this dose cortisol levels also rose in the pregnant animals (baseline = 1040 +/- 30 nmol/L; peak, 1620 +/- 130); however, this response was blunted compared to that in the nonpregnant animals (P less than 0.05). CRH (5.0 micrograms/kg) significantly stimulated both ACTH and cortisol in the nonpregnant animals.(ABSTRACT TRUNCATED AT 400 WORDS)     

Correlation between plasma levels of ACTH and beta-endorphin in the first seven days of postnatal life

Plasma adrenocorticotropic hormone (ACTH) and beta-endorphin (beta-EP) concentrations were measured by radioimmunoassay in 122 newborns, born vaginally after spontaneous labor between the 38th and the 41st week of pregnancy. Blood samples were taken from umbilical cord in 10 newborns and from antecubital vein in the remaining 112 newborns, divided into 8 groups aged from 12 h to 7 days. The mean (+/- SE) ACTH concentrations in cord plasma were 81.87 +/- 10.16 pg/ml and decreased significantly (p less than 0.01) at the 24th h of life (49.09 +/- 6.93 pg/ml). Afterwards mean ACTH plasma concentrations fluctuated around the latter values. The mean (+/- SE) B-EP concentrations in cord plasma were 21.96 +/- 3.12 pmol/l and decreased significantly at the 24th h of life (13.43 +/- 2.08 pmol/l; p less than 0.01). From the 24th h to the 7th day the mean plasma concentrations of beta-EP were not significantly different. ACTH and beta-EP plasma levels were positively correlated (p less than 0.001) at delivery and during the first seven days of life.     

Adrenomedullin production is increased in normal human pregnancy

OBJECTIVE: Adrenomedullin, a recently discovered vasoactive peptide originally identified in pheochromocytoma, has been found to be increased in the plasma of pregnant women at term. This study was designed to elucidate whether adrenomedullin secretion is dependent on gestational age and the possible source and function of this peptide in human pregnancy. STUDY DESIGN: Adrenomedullin concentrations were determined by RIA in amniotic fluid and maternal plasma obtained from 110 pregnant women between 8 and 40 weeks of gestation. Subjects were stratified into five groups according to gestational age. In term patients (n = 15), adrenomedullin was also measured in the umbilical artery and vein separately. RESULTS: High concentrations of adrenomedullin were present in plasma and amniotic fluid samples from patients in the first, second and third trimester. There was no significant difference in mean maternal plasma concentration of adrenomedullin between the five patient groupings. Amniotic fluid adrenomedullin concentrations decreased from 81.2 +/- 11.7 pg/ml at 8-12 weeks of gestation to 63.7 +/- 6.0 pg/ml at 13-20 weeks of gestation and then increased at 21-28 weeks of gestation to 99.1 +/- 10.4 pg/ml. A further increase was found in samples collected after 37 weeks of gestation (132.6 +/- 10.1 pg/ml). In the umbilical vein, adrenomedullin concentration was higher (P < 0.05) than in the artery (65.7 +/- 6.1 pg/ml and 48.5 +/- 5.2 pg/ml respectively), suggesting that adrenomedullin in the fetal circulation derives from the placenta. CONCLUSIONS: Our results demonstrate the presence of adrenomedullin in maternal plasma and amniotic fluid throughout gestation, and show that its production starts very early in gestation, suggesting that this hormone may have an important role in human reproduction, from implantation to delivery.     

Atrial natriuretic peptide in umbilical cord blood: evidence for a circulating hormone in human fetus

To determine whether atrial natriuretic peptide (ANP) is a circulating hormone in the human fetus, ANP levels in umbilical cord plasma obtained at the time of delivery in 10 normal infants were measured by RIA. Plasma ANP levels were consistently higher in paired umbilical cord arterial than in cord venous samples. The mean umbilical cord arterial plasma ANP concentration [283 +/- 56 (+/- SEM) pg/ml] was significantly higher than that in cord venous plasma (165 +/- 27 pg/ml) or maternal peripheral venous plasma (155 +/- 25 pg/ml). Analyses by reverse phase high performance liquid chromatography revealed that the elution pattern of plasma ANP in cord arterial blood and that in maternal venous blood were nearly identical and that the retention time of the main ANP peak coincided with that of alpha-human ANP. These results suggest that alpha-human ANP is a circulating hormone in the human fetus.     

Plasma corticotropin-releasing factor concentrations in the baboon during pregnancy.

We have studied the secretion of placental CRF during pregnancy in the baboon, an animal model with many similarities to human pregnancy. Plasma CRF was measured in two groups of animals. In group 1, studies were performed in six anesthetized animals beginning 8 days postconception. In group 2, studies were performed in five unanesthetized chronically catheterized maternal and five fetal animals in the latter third of pregnancy. In the first study beginning early in pregnancy, CRF was undetectable in all animals on days 8 and 15 postconception. Plasma CRF became detectable in two animals on day 24 and in the remaining four on day 30. Plasma CRF rose significantly to a mean of 810 +/- 160 pg/ml at 37 days gestation (F = 4.20; P < 0.001). Mean maternal plasma CRF was 2452 +/- 1120 pg/ml on day 44 and remained elevated, with a great deal of variability between subjects, until the end of the study period (128 days of gestation). Samples in this group were obtained after ketamine sedation. The effect of ketamine on CRF was studied in three chronically catheterized animals. Samples were obtained before and 2, 4, 6, and 24 h after ketamine administration (40 mg, iv). The baseline CRF concentration was 1168 +/- 131 pg/ml and did not change significantly over the time period studied. In the second study in the chronically catheterized animals, maternal plasma CRF was 1990 +/- 680 pg/ml at 131-140 days gestation and remained elevated until near term at 170 days (term = 175-180 days). Within 24 h after birth, plasma CRF became undetectable (< 60 pg/ml). CRF was also measured in chronically catheterized fetal baboons. The mean CRF concentration was 614 +/- 224 pg/ml at 131-140 days and remained in this range until the end of the period studied (151-160 days gestation). To characterize the CRF immunoactivity in maternal baboon plasma, Sephadex chromatography was performed on an 8.4-ml plasma sample obtained at 160 days gestation. The majority of the CRF immunoactivity eluted in the same position as synthetic human CRF. We conclude that high levels of placental CRF are present in the systemic circulation of the maternal and fetal baboon during pregnancy. In contrast to human pregnancy, which is characterized by an exponential rise in maternal CRF concentrations in the final weeks before delivery, an exponential rise in maternal baboon CRF concentrations occurs early in pregnancy.     

Amniotic fluid and umbilical cord plasma corticotropin-releasing factor (CRF), CRF-binding protein, adrenocorticotropin, and cortisol concentrations in intraamniotic infection and inflammation at term

Context: Pregnant tissues express corticotropin-releasing factor (CRF), a peptide modulating fetal and placental ACTH and cortisol secretion. These actions are modulated by the locally expressed CRF-binding protein (CRF-BP). Objective: The objective of the study was to determine whether CRF, CRF-BP, ACTH, and cortisol concentrations change in amniotic fluid and umbilical cord plasma in the presence of intraamniotic infection/inflammation (IAI) in women with spontaneous labor at term. Design: This was a cross-sectional study. Setting: The study was conducted at a tertiary referral center for obstetric care. Patients: Patients included women in active labor at term with (n = 39) and without (controls; n = 78) IAI. Main Outcome Measures: Amniotic fluid and umbilical cord plasma concentrations of CRF, CRF-BP, ACTH, and cortisol measured by RIA and immunoradiometric assays were measured. Results: In patients with IAI, amniotic fluid CRF (0.97 +/- 0.18 ng/ml) and CRF-BP (33.06 +/- 5.54 nmol/liter) concentrations were significantly (P < 0.001) higher than in controls (CRF: 0.32 +/- 0.04 ng/ml; CRF-BP: 14.69 +/- 2.79 ml). The umbilical cord plasma CRF and CRF-BP concentrations were significantly (P < 0.001 for all) higher in women with IAI than in controls (CRF: 2.96 +/- 0.35 ng/ml vs. 0.38 +/- 0.18 ng/ml; CRF-BP: 152.12 +/- 5.94 nmol/liter vs. 106.9 +/- 5.97 nmol/liter). In contrast, amniotic fluid and umbilical cord plasma ACTH and cortisol concentrations did not differ between groups. Conclusions: Amniotic fluid and umbilical cord plasma CRF and CRF-BP concentrations are increased in women with spontaneous labor at term and IAI. CRF-BP may modulate CRF actions on ACTH and cortisol secretion, playing a pivotal role in limiting the inflammatory process and thus avoiding an overactivation of the fetal/placental hypothalamus-pituitary-adrenal axis at birth.     

Oestrone, oestradiol-17beta and oestriol levels in human foetal plasma during gestation and at term

Marked rises in both unconjugated and sulphoconjugated estrone, estradiol-17-beta and estriol were observed in human fetal plasma between midgestation and term. Significant arterio-venous differences were found in the umbilical cord plasma. No consistent arterio-venous differences were found in the umbilical cord plasma. This indicates that all 3 estrogens are secreted from the placenta into the fetal circulation in the unconjugated form. Mean unconjugated estrogen (estrone + estradiol-17-beta + estriol) levels rose from 22.7 ng/ml at 17-20 weeks of gestation to 108.9 ng/ml at term in umbilical venous plasma and from 4.3 ng/ml to 23.3 ng/ml in umbilical arterial plasma. This represents a secretion rate of approximately 30 mg estrogen/day into the umbilical vein at term. Mean estrogen sulphate levels rose from 128 ng/ml to 313 ng/ml in the cord plasma during the same period. Of the 3 estrogens measured, estriol was quantitatively the major estrogen in fetal plasma. It consistently represented about 78% of the unconjugated fraction and 95% of the sulphate fraction at all stages of gestation. The method of delivery did not have a significant effect on the estrogen levels in uncomplicated pregnancies. Similar estrogen levels were found in fetal heart blood after either hysterotomy at spontaneous abortion at 16-20 weeks of gestation, and no significant differences were found for estrogen levels in cord plasma after elective Caesarean section at 38-39 weeks when compared with estrogen levels after normal delivery at term. A significant rise in fetal unconjugated estrogens at a time when fetal corticosteroids are increasing may be of physiological importance for fetal maturation in women.     

Plasma beta-endorphin and beta-lipotropin in the human fetus at delivery: correlation with arterial pH and pO2.

Beta-endorphin-like immunoactivity was measured in the umbilical cord plasma of 45 term human fetuses. Mean concentration was 91 +/- 16 (SEM) pg/ml,an the normal adult level of 30.7 +/- 2.7 pg/ml. This immunoactivity was further characterized in 10 cases by Sephadex G-50 chromatography to separate beta-endorphin from beta-lipotropin (beta-LPH). Mean beta-endorphin and beta-LPH concentrations were 57 +/- 12.8 and 455 +/- 101 pg/ml, respectively. Both were higher (P less than 0.01) than the mean beta-endorphin and beta-LPH concentrations reported in the adult. The mean molar beta-endorphin to beta-LPH ratio was 0.35 in the fetus and 0.36 in the adult. In 17 fetuses whose umbilical arterial and venous concentrations were measured separately, mean beta-endorphin-like immunoactivity was higher in the artery than in the vein. A highly significant negative correlation (r = -0.831; P less than 0.001) was present between umbilical arteiral pH and beta-endorphin-like immunoactivity. A negative correlation (r = -0.611; P less than 0.005) with arterial pO2 was also noted. We conclude that high levels of beta-endorphin-like immunoactivity, composed of both beta-endorphin and beta-LPH, circulate in the human fetus at term, and that hypoxia and secondary acidosis may be major stimuli to the release of these peptides.     

Difference of plasma leptin levels in venous and arterial cord blood: relation to neonatal and placental weight

To investigate the physiological significance of umbilical plasma leptin in the fetal growth and its possible origin of production during pregnancy, plasma leptin concentrations in venous and arterial cord bloods were measured in 42 neonates (male = 23, female = 19, gestational age 36-41 weeks, birth weight 2600-4000 g). Leptin concentrations in umbilical veins (5.65 +/- 0.53 ng/mL, n = 42) and umbilical arteries (0.56 +/- 0.07 ng/mL, n = 42) were significantly (P < 0.001) lower than those in maternal peripheral veins (22.36 +/- 1.06 ng/mL, n = 42). Mean plasma leptin concentration (+/- SEM) in venous cord blood was significantly (P < 0.001) higher than that of arterial cord blood. There was significantly positive correlation (r = 0.447, P < 0.01) between umbilical venous leptin levels and neonatal body mass index (BMI). A positive correlation (r = 0.435, P < 0.01) was also found between umbilical arterial leptin and neonate BMI. There was no positive correlation between umbilical leptin levels in venous cord blood and placental weight. We thus conclude that umbilical plasma leptin may play an essential role in the fetal growth and metabolism during pregnancy and placenta is one of the major sources of leptin production, but the level of leptin in umbilico-placental circulation is independent of the weight of placenta.     

Relationship between maternal and fetal corticotrophin-releasing hormone-41 and ACTH levels in human mid-trimester pregnancy

Samples of maternal blood, amniotic fluid and umbilical arterial and venous blood were collected from 11 women at 16-24 weeks of pregnancy. Corticotrophin-releasing hormone-41 (CRH-41) and ACTH were measured by immunoradiometric assay. The mean levels of ACTH were 11 pmol/l in maternal plasma, 12 pmol/l in fetal plasma and 9.7 pmol/l in amniotic fluid. The mean levels of CRH-41 were 1.6 pmol/l in maternal plasma and 0.7 pmol/l in fetal plasma. There was a positive correlation between maternal and fetal plasma CRH-41 and between maternal CRH-41 and ACTH. In fetal plasma there was a weak inverse correlation between CRH-41 and ACTH. This is the first demonstration of CRH-41 in the circulation of the mid-trimester human fetus, but on the basis of the present findings it is not possible to specify the exact source (fetal, placental or maternal).     

Plasma adrenocorticotropin and cortisol concentrations during acute hypoxemia after a reversible period of adverse intrauterine conditions in the ovine fetus during late gestation

The present study determined the pituitary-adrenal responses to acute hypoxemia after a period of reversible adverse intrauterine conditions produced by partial compression of the umbilical cord for 3 days in the sheep fetus during late gestation. At 118 +/- 2 days gestation (term is approximately 145 days), 12 sheep fetuses were instrumented under halothane anesthesia with an occluder cuff around the umbilical cord, amniotic and vascular catheters, and a transit-time flow probe around an umbilical artery. In 6 of the fetuses at 125 days, umbilical blood flow was reduced by about 30% from baseline for 3 days (UCC), after which the occluder was deflated. The remaining 6 fetuses acted as sham-operated controls in which the occluder was not inflated. All fetuses were then subsequently subjected to 2 periods of acute hypoxemia, elicited by reducing the maternal inspired fraction of oxygen (FiO(2)) at 2 +/- 1 and 5 +/- 2 days after the end of cord compression or sham compression. In addition, 4 fetuses from each group were subjected to an ACTH challenge 1-2 days after the final episode of acute hypoxemia. Maternal and fetal arterial blood samples were taken at appropriate intervals during cord compression, acute hypoxemia, and ACTH challenge for analyses of blood gases, pH, and plasma ACTH and cortisol concentrations. Partial compression of the umbilical cord produced reversible mild fetal asphyxia, a transient increase in fetal plasma ACTH, and a progressive increase in fetal plasma cortisol. At 5 +/- 2 days after the end of compression, despite similar blood gas status between the groups, basal plasma cortisol, but not ACTH, concentrations were significantly greater in compressed fetuses relative to sham controls. However, this dissociation did not affect a similar increment in fetal plasma ACTH and cortisol concentrations during acute hypoxemia or in the fetal plasma cortisol response to the ACTH challenge in either group. An increase in adrenocortical mass occurred in fetuses preexposed to partial compression of the umbilical cord relative to sham controls. The data suggest that fetal exposure to a reversible period of adverse intrauterine conditions produced by partial compression of the umbilical cord does not affect the magnitude of the fetal hypothalamic-pituitary-adrenal axis response to subsequent acute hypoxemia, but it leads to resetting of basal hypothalamic-pituitary-adrenal axis function in the fetus. The mechanism for this resetting may include an increase in adrenocortical steroidogenic synthetic capacity, but it is not due to a change in adrenocortical sensitivity to ACTH. Inappropriate fetal glucocorticoid exposure after reversible periods of adverse intrauterine conditions has important implications for fetal and postnatal development.     

Characterization of plasma beta-endorphin immunoactivity in the fetal lamb: effects of gestational age and hypoxia

To characterize the immunological forms of beta-endorphin (beta-EP) in the fetal circulation, total beta-EP immunoactivity [beta-EPi] and N-acetyl beta-EPi were measured in the plasma of chronically catheterized fetal lambs in undisturbed conditions and before, during, and after periods of controlled hypoxia. Measurements of the peptide concentrations in each plasma sample were made by RIA using an antiserum to the midportion of beta-endorphin which cross-reacts with both acetylated and unacetylated forms of the peptide as well as with beta-lipotropin, and a second antiserum which reacts only with acetylated forms of beta-EP. In 25 plasma samples from 12 fetal animals at 113-142 days gestation, total beta-EPi was 87.0 +/- 10.9 pg/ml, while N-acetyl beta-EPi was 90.8 +/- 7.7 pg/ml (mean +/- SE). When a plasma pool obtained from 3 fetuses in the basal state was extracted and chromatographed on Sephadex G-50, most of the N-acetyl beta-EPi eluted in the same position as the synthetic N-acetyl beta-EP standard. Thus, most of the beta-EPi in the plasma of the unstressed fetus could be accounted for by N-acetylated forms of the peptide. These are the major forms of beta-EP produced by the intermediate lobe of the pituitary. To examine the effects of acute hypoxia on fetal plasma peptide levels, pregnant ewes were exposed to 10% O2 in N2 for 30 min. In 15 studies at 113-142 days gestation, mean fetal PO2 decreased from 21.7 +/- 0.6 to 11.0 +/- 0.7 mm Hg (P less than 0.001). Total beta-EPi increased significantly from 93.0 +/- 17.7 to 527 +/- 146 pg/ml during hypoxia and returned toward basal values after 30 min of recovery to 372 +/- 116 pg/ml (P less than 0.02). Over the same intervals, N-acetyl beta-EPi did not change significantly, with mean levels of 88.5 +/- 10.7, 123 +/- 16.3, and 130 +/- 16.8 pg/ml. This shows that the increase in total beta-EPi with hypoxia could not be accounted for by an increase in N-acetyl beta-EPi. Our finding that most of the total beta-EPi in the circulation of the undisturbed fetus is N-acetyl beta-EPi favors an intermediate lobe origin. Since beta-EP is inactivated by N-acetylation, these data suggest that this immunoactivity has little or no biological activity. Enhanced release of total beta-EPi during hypoxia, which could not be accounted for by acetylated forms, suggests that this type of stress activates the anterior pituitary lobe and results in increased plasma concentrations of the biologically active peptide.     

Corticotropin-releasing hormone and oxytocin stimulate the release of placental proopiomelanocortin peptides

Human placenta contains the POMC-derived peptides ACTH, alpha MSH, and beta-endorphin, and CRH. High concentrations of immunoreactive (IR) CRH have been recently demonstrated in maternal plasma during pregnancy. To determine if the human placenta secretes CRH and POMC-derived peptides, we developed an in vitro human placental fragment perifusion system. The perifused tissue released IR-CRH and POMC-derived peptides at a constant rate for at least 5 h. The mean IR-CRH concentration in the effluent (under basal conditions) was 158 +/- 26 (+/- SD) pg (34.5 +/- 5.8 fmol)/5-min fraction.g tissue. IR-alpha MSH, IR-beta-endorphin, and IR-ACTH were also released into the perifusion medium; the mean concentration of alpha MSH released was 24.6 +/- 8 pg (14.8 +/- 4.8 fmol)/fraction.g, that of ACTH was 2.9 +/- 1.9 pg (0.65 +/- 0.43 fmol)/fraction.g, and that of beta-endorphin was 12.9 +/- 6 pg (3.8 +/- 1.7 fmol)/fraction.g. We examined the effects of human CRH, oxytocin, vasopressin, and dexamethasone on placental POMC peptide secretion. Five-minute pulses of 10(-8) or 10(-6) mol/L human CRH or oxytocin produced an immediate and dose-dependent increase in all POMC peptides in the effluent. A 5-min pulse of 10(-8) or 10(-6) mol/L vasopressin had no effect. A continuous 4-h exposure to 10(-6) mol/L dexamethasone had no effect on either basal IR-CRH or POMC-derived peptide or their KCl-induced release. In conclusion, we found that 1) human placenta releases IR-CRH and POMC-derived peptides in vitro; this phenomenon seems to be independent of glucocorticoid control; 2) placental CRH may have a paracrine effect on placental POMC peptide release in addition to its possible action on maternal pituitary hormone release; and 3) oxytocin, but not vasopressin, stimulates placental POMC peptide release.     

Regulation of basal adrenocorticotropin and cortisol secretion by arginine vasopressin in the fetal sheep during late gestation.

This study tested the hypothesis that arginine vasopressin (AVP) is involved in the regulation of basal ACTH secretion in the ovine fetus near term. In five fetuses challenged with AVP (1 microgram/ml, iv bolus) plasma ACTH concentrations increased to an 8-fold peak within 10 min of the preceding baseline (55 +/- 6 to 403 +/- 241 pg/ml). Cortisol in fetal circulation subsequently increased 2-fold (11 +/- 1 to 28 +/- 5 ng/ml) within 15 min of the AVP injection. The AVP-induced rise in plasma ACTH and cortisol concentrations was blocked when the fetus was pretreated with the AVP V1 receptor antagonist d(Ch2)5Tyr(Me)AVP. In a total of seven studies, antagonist (10 micrograms/kg estimated BW, iv bolus) was administered to three fetuses, aged 137-147 days gestation, followed 40 min later by the exogenous AVP challenge, as described above. After AVP antagonist treatment, basal ACTH and cortisol concentrations were not significantly different from the preinjection baseline levels (P greater than 0.05, by analysis of variance). Moreover, plasma ACTH and cortisol remained unchanged after the AVP challenge. To further define the role of endogenous AVP in basal ACTH and cortisol secretion, the AVP antagonist was administered (five studies in two fetuses) at 30-min intervals for a total of three injections per fetus. This extended AVP antagonist regimen also failed to alter fetal circulating concentrations of ACTH or cortisol (P greater than 0.05). Cortisol in the maternal circulation was not affected by any of the fetal AVP or AVP antagonist treatments. Lambs were born at 146 +/- 2 days gestation (n = 5), within the range for the normal duration of pregnancy. These data do not support the hypotheses that AVP is involved in the regulation of basal ACTH secretion in the fetal sheep during the 10 days preceding parturition. Rather, the ability of AVP antagonist to block the AVP-induced rise in plasma ACTH and cortisol in the fetus suggests that basal and stimulated ACTH secretion are under separate regulatory mechanisms.     

Responsivity of the baboon fetal pituitary to corticotropin-releasing hormone in utero at midgestation.

The present study was designed to determine whether the baboon fetal pituitary at midgestation was responsive in utero to a bolus injection of CRH. On day 100 of gestation (term = day 184), baboons were anesthetized with halothane/nitrous oxide, the fetus was exteriorized, and a cannula was inserted into a fetal carotid artery. Five minutes later (experimental time zero), a fetal carotid blood sample was obtained, and saline (0.5 ml) with (n = 6) or without (n = 3) ovine CRH (100 ng estimated to equal 500 ng/kg BW) was then infused via the fetal carotid over a 3-min period. Fetal blood samples were taken 5, 15, 30, 45, and 60 min post-CRH/saline treatment and assayed for ACTH. Mean (+/- SE) pretreatment fetal plasma ACTH concentrations were similar in animals that subsequently received saline (26 +/- 3 pg/ml) or CRH (29 +/- 6 pg/ml). Fetal plasma ACTH remained constant after the infusion of saline. In contrast, CRH increased (P less than 0.05) fetal plasma ACTH within 5 min in six of six baboons to a value (58 +/- 12 pg/ml) that exceeded (P less than 0.05) the zero time value and the respective mean value (27 +/- 5 pg/ml) in saline-treated fetuses. Fetal plasma ACTH concentrations continued to rise in four of six baboons 15 min after CRH injection to a level (68 +/- 15 pg/ml) which exceeded that in saline controls (27 +/- 2 pg/ml). In fetuses treated with CRH, overall mean fetal plasma ACTH concentrations from 0-60 min increased at a rate (1.47 pg/min) greater (P less than 0.05) than that in fetuses injected with saline (0.07 pg/min). In contrast to the effects of intracarotid CRH injection, fetal plasma ACTH was not increased after the infusion of 100 ng CRH into a fetal antecubital vein of three additional animals. Collectively, these findings indicate that intracarotid injection of a bolus of CRH into the baboon fetus rapidly increased fetal plasma ACTH concentrations. Moreover, the site of action of CRH was presumably the fetal pituitary. Therefore, we suggest that the baboon fetal hypothalamic-pituitary axis at midgestation has the capacity to secrete ACTH in response to a challenge of CRH.     

Corticotrophin-releasing hormone and ACTH levels in maternal and fetal blood during spontaneous and oxytocin-induced labour

The changes in corticotrophin-releasing hormone (CRH), ACTH and dehydroepiandrosterone (DHEA) in maternal and fetal plasma were estimated in women undergoing spontaneous and oxytocin-induced labour to correlate hormone changes with the mode of parturition. Blood was sampled from a maternal peripheral vein 2 days before labour, during the second stage of labour and on the second postnatal day, and also from umbilical vessels just after delivery. Hormone concentrations were measured by RIA and ELSA methods. The maternal plasma CRH concentration before labour was significantly higher in the group of women delivered spontaneously and declined during the labour through to the second postnatal day. Measured in umbilical vessels, CRH as well as ACTH concentrations were higher in the umbilical vein than artery. The mean maternal plasma ACTH was similar in both groups before delivery, then increased significantly in both groups during the labour, decreasing on the second day after delivery. There were no changes in DHEA concentrations among the groups and at all time points of collection. No correlations between CRH and ACTH or DHEA were observed. Our results suggest that the maternal pituitary can respond to stress factors during delivery but peripheral CRH, probably mainly of placental origin, is not a major modulator of pituitary action.     

The newly developed method of 19-OH-A as intermediate of estrone biosynthesis by GC-MS

To study the serum levels of 19-hydroxyandrostenedione (19-OH-A), known as an obligatory intermediate of estrogen biosynthesis and considered to be one of the hypertensinogens, a method using GC-MS with application of synthesized [7,7-d2]androstenedione (A), [7,7-d2] 19-OH-A and [9,11-d2]estrone(E1) as internal standards was newly developed. Normal pregnant women and pregnant women complicated with hypertension near term were selected for the study. The levels of 19-OH-A and E1 increased significantly as gestation progressed [19-OH-A; 224.7 +/- 72.1 (1st trimester), 655.5 +/- 325.4 (2nd trimester). 1517.8 +/- 543.6 (3rd trimester)pg/ml], and a positive correlation was found between the levels of the two steroids. No apparent change was observed in A levels during the course of pregnancy. The mean levels of 19-OH-A in pregnancy complicated with hypertension at 2nd and 3rd trimester were 761.7 +/- 348.9 and 1473.0 +/- 491.4 pg/ml, which were compatible with those in normal pregnancy. The levels of 19-OH-A at delivery in maternal vein (MV) were 1735.1 +/- 683.9 pg/ml. Significantly higher levels of 19-OH-A were found in umbilical vein (UV) (1977.2 +/- 564.9 pg/ml) than those in umbilical artery (109.7 +/- 49.1 pg/ml). 19-OH-A concentration in term placental tissue was 16.3 ng/g.w.w. tissue. This is the first report to demonstrate the serum 19-OH-A levels measured by GC-MS and also to demonstrate the levels in the cord blood. The results indicate that 19-OH-A may be the product of pregnancy and may be derived from the feto-placental compartment.