Biomedical subjects
Publications and source records attributed to B R Carr.
Discussed in this article are the effects of pregnancy-induced hypertension on the renin-angiotensin-aldosterone system; the possible role of prostanoids, deoxycorticosterone, and deoxycorticosterone sulfate in blood pressure control; and the effect of pregnancy-induced hypertension upon the endocrinology of the maternal-fetal-placental unit.
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The weights of kidney obtained from 200 presumably normal human abortuses of postconceptional gestational ages between 6 and 17 weeks were measured. The weight of the normal human fetal kidney increases in an exponential manner between 6 and 14 weeks gestation. Thereafter, the rate of increase in weight is slower. No significant differences were found between the weights of kidneys of male and female abortuses of 7-15 weeks postconceptional gestational ages.
The weights of 182 adrenal glands obtained from presumably normal human fetuses delivered by elective abortion between 6 and 17 weeks post-conceptional gestational age were measured. There was little increase in adrenal weight between 6 and 12 weeks gestation. Thereafter, the rate of increase in adrenal weight was rapid. There were no significant differences between the adrenal gland of male and female abortuses of similar gestational ages.
The binding of [125I]iodoLDL and [125I]iodoHDL to membrane fractions prepared from fresh adrenal tissue of an anencephalic newborn was examined and compared to the binding of these lipoproteins to membrane fractions prepared from normal human fetal adrenal (HFA) tissue. The apparent dissociation constant (KD) for the interaction of [125I]iodoLDL with its binding sites in membrane fractions prepared from HFA tissue of an anencephalic fetus (23.5 micrograms . ml-1) was similar to that in membranes from normal HFA tissue. However, the binding capacity for [125I]iodoLDL of membrane fractions prepared from adrenal tissue of an anencephalic fetus (600 ng . mg-1 protein) was only one-third of the binding capacity (1500 ng . mg-1 protein) of normal HFA membrane fractions. On the other hand, the binding capacity for [125I]iodoHDL of membrane fractions prepared from anencephalic and normal adrenal tissue was similar.
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Human fetal adrenal (HFA) tissue was maintained in organ culture to evaluate the biosynthesis of prostaglandins and hormonal regulation of prostaglandin formation by this tissue. The HFA tissue secreted substantial amounts of prostaglandin E(2), prostaglandin F(2alpha), 13,14-dihydro-15-ketoprostaglandin F(2alpha), 6-ketoprostaglandin F(1alpha), and thromboxane B(2); secretion of prostaglandin D(2) could not be demonstrated. Prostaglandin biosynthesis in HFA tissue was inhibited in a time-dependent manner by corticotropin (ACTH; 0.4 muM); by the fourth day of culture, the extent of inhibition of biosynthesis of each prostaglandin was 60-90%. Progesterone (1 muM), cortisol (1 muM), and dexamethasone (1 muM) inhibited prostaglandin biosynthesis whereas estradiol (1 muM) did not. Of the compounds tested for inhibitory activity, dexamethasone was the most potent. An inhibitor of 11beta-hydroxylase activity (metyrapone; 0.1 mM) effectively eliminated the inhibition of prostaglandin biosynthesis caused by corticotropin and progesterone. Metyrapone treatment alone caused a 3-fold increase in prostaglandin biosynthesis by fetal adrenal tissues. Similar stimulatory effects resulted from treatment with inhibitors of (i) 3beta-hydroxysteroid dehydrogenase (cyanoketone; 15 muM), (ii) steroid 17alpha-hydroxylase (SU 10603; 19 muM), and (iii) cholesterol side-chain cleavage (aminoglutethimide; 1 mM). Inhibition of prostaglandin biosynthesis by dexamethasone in the presence or absence of metyrapone was concentration dependent and 50% inhibition could be demonstrated at 1 nM. A competitive inhibitor of the binding of glucocorticosteroids to cytoplasmic receptors (cortisol 21-mesylate; 1 muM) significantly reduced the inhibition of prostaglandin biosynthesis effected by dexamethasone (10 nM). These findings suggest that prostaglandin biosynthesis in the HFA gland is regulated by endogenously synthesized glucocorticosteroids, the actions of which are mediated by a glucocorticosteroid receptor. Such glucocorticosteroids induce the synthesis of a substance that inhibits prostaglandin biosynthesis.
The present study was undertaken to define the umbilical cord plasma concentrations of cholesterol throughout human gestation. Mixed arterial and venous cord plasma samples obtained from abortuses of women undergoing elective abortion or from infants of women who underwent spontaneous premature vaginal delivery, and from infants of women who delivered vaginally at term were assayed for cholesterol by a micro-enzymatic method. No cases that involved any maternal or fetal complications (other than prematurity) were included in this study. Early in gestation (10-16 weeks post-conception), the total cholesterol level in cord plasma was 85.4 +/- 30.7 mg/dl (mean +/- SD), N = 68, with the cholesterol levels in some samples falling within the range of those of adults. Between 16.5 and 20 weeks post-conception, the umbilical cord plasma cholesterol level declined to 39.9 +/- 21.0 mg/dl, n = 19 (P less than 0.001). The cholesterol concentration in umbilical cord plasma then rose to 67.8 +/- 24.0 mg/dl, n = 17 (P less than 0.01) between 26.5 and 32 weeks of gestation. Thereafter, a second decline in the umbilical cord plasma cholesterol level occurred, with the values at 32.5-36 weeks being 58.4 +/- 13.6 mg/dl (n = 16), and at 36.5 to 40 weeks post-conception (term) being 51.4 +/- 11.5 mg/dl, n = 44 (P less than 0.01 vs. 26.5-32 wks). We suggest that the observed changes in fetal cholesterol levels could be related to alterations during development in the rates of lipoprotein-cholesterol biosynthesis and subsequent clearance from plasma by the fetal adrenals wherein cholesterol is used as substrate for steroid biosynthesis.
In a previous report evidence was presented that plasma low density lipoprotein (LDL), but not high density lipoprotein (HDL), is the major source of cholesterol used by the human corpus luteum for progesterone biosynthesis and, that plasma LDL is taken up by corpus luteum tissues via a receptor-mediated endocytotic process. Using membrane fractions prepared from fresh corpus luteum tissue obtained from nine women at various phases of the menstrual cycle, we conducted the present investigation to characterize lipoprotein-binding sites and changes in such binding sites that might occur throughout the ovarian cycle. High affinity, low capacity binding sites for [125I]iodo-LDL and also for [125I]iodo-HDL were detected in membrane fractions prepared from fresh corpus luteum tissue. The interaction of [125I]iodo-LDL with the high affinity binding sites in fresh corpus luteum membrane fractions was prevented by incubation with heparin. Also, the binding capacity of corpus luteum membrane fractions for [125I]iodo-LDL was reduced by treatment with pronase. On the other hand, the specific binding capacity for [125I]iodo-HDL was unaffected by pronase treatment. The binding capacity for [125I]iodo-LDL in membrane fractions prepared from midluteal phase corpus luteum was significantly greater than that of membrane fractions from tissue obtained at any other phase of the cycle, a finding that suggests that changes in progesterone biosynthesis throughout the cycle are positively correlated with changes in the numbers of binding sites for LDL in the corpus luteum.
The binding of low density lipoprotein (LDL) and the de novo synthesis of cholesterol in separated zones of human fetal adrenal (HFA) tissues were investigated. The number of LDL-binding sites was 2-fold greater in membrane fractions prepared from fresh fetal zone tissue than in those from neocortex tissue. The binding capacity for LDL in fetal zone and neocortex membrane preparations of HFA tissues maintained in culture in the presence of ACTH was 2-fold greater than that in membrane fractions of control tissues. The rates of de novo synthesis of cholesterol also were determined in separated zones of HFA tissue by measuring the specific activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase in microsomal fractions prepared from HFA tissues and by determining the rate of incorporation of tritium from [3H]water into cholesterol in HFA tissue fragments. The rate of de novo synthesis of cholesterol in fresh fetal zone tissue was twice that in neocortex tissue as estimated by these methods. When separated zones of HFA tissue were maintained in culture in the presence or absence of ACTH, the rates of de novo synthesis, as determined by the rate of incorporation of tritium from [3H]water into cholesterol, were stimulated to a similar extent by ACTH in both fetal zone and neocortex tissues. However, the specific activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase was increased to a greater extent by ACTH pretreatment in neocortex tissues than in fetal zone tissues. In summary, fetal zone tissues of the HFA gland have a larger number of LDL-binding sites and higher rates of de novo synthesis of cholesterol than do neocortex tissues, and ACTH stimulates LDL binding and de novo synthesis of cholesterol in both zones of the HFA gland.
In the current investigation we sought to evaluate fetal adrenal dehydroepiandrosterone sulfate (DS) production during normal human pregnancy. We found that the mean umbilical cord plasma concentration of DS was reasonably constant from 18-34 weeks of gestation, ranging from 1131-1517 ng/ml. Thereafter, the cord plasma DS concentration rose steadily to 2463 +/- 665 ng/ml (mean +/- SD) at 39-40 weeks of gestation. Between 20 and 32 weeks of gestation, the estimated fetal plasma DS pool increased from 8 to 146 microgram. At term, the estimated fetal DS plasma pool was in excess of 300 microgram. From 34-40 weeks of gestation, the estimated fetal DS pool size and the fetal plasma DS concentration increased coincidentally with a marked rise in the known rate of fetal adrenal growth. From these findings, we conclude that fetal adrenal DS production increases steadily throughout gestation, and the rate of increase is probably dependent upon the rate of fetal adrenal growth. Furthermore, the rapid increase in maternal estrogen levels near term observed by numerous investigators can be explained in part by an increased availability of the fetal estrogen precursor DS at this time.
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The characteristics of binding of hCG to testicular tissue obtained from human abortuses of 10--24 weeks gestational age were studied. Specific, saturable binding of [125I]hCG was demonstrated using homogenates of human fetal testicular tissue. The equilibrium dissociation constant ranged from 0.4 x 10(-10) M to 5.5 x 10(-10) M, a finding that is indicative of a high affinity receptor. The capacity to bind hCG was low, but varied strikingly with gestational age. The binding capacity for hCG of tissues from abortuses of gestational age less than 15 weeks and greater than 22 weeks was consistently less than 10.0 pg x mg-1 tissue (2.2 fmol x mg-1 tissue). The binding capacity for hCG of tissues from abortuses of gestational age between 15--20 weeks ranged from 2.4--29.8 pg x mg-1 tissue (0.5--6.5 fmol x mg-1 tissue) with the majority of values being greater than 10 pg x mg-1 tissue (2.2 fmol x mg-1 tissue). On the other hand, receptors for hCG in human fetal ovarian tissue were undetectable, irrespective of gestational age. It is concluded that specific high affinity binding sites for hCG are present in human fetal testes and that the binding capacity is maximum between gestational ages of 15--20 weeks. This increase in binding capacity parallels the surge in testosterone production known to occur during the same period of development. These results suggest that the increase in fetal plasma levels of testosterone during this time in gestation is the result of an increase in the sensitivity of the fetal testis to hCG caused by an increase in the number of hCG receptors, and that hCG most likely is responsible for stimulation of fetal testicular steroidogenesis in utero at this time of gestation.
Adrenocorticotropin (ACTH) and cortisol in plasma were measured weekly from early in gestation through delivery in five women whose pregnancies were normal. During the twelfth week of pregnancy, the concentration of ACTH in plasma of blood samples obtained between 0800 and 0900 hours was 23 +/- 4.6 pg/ml (mean and SEM) and rose progressively to 59 +/- 16 pg/ml at 37 weeks. The levels of ACTH in plasma were significantly lower throughout pregnancy than those found in nonpregnant women. During labor and delivery, ACTH levels rose strikingly to values of 301 +/- 137 pg/ml. As pregnancy advanced, the concentration of cortisol in plasma increased progressively from 149 +/- 34 ng/ml (mean and SEM) at 12 weeks to 352 +/- 90 ng/ml at 26 weeks' gestation but changed minimally thereafter until labor commenced, during which values of 706 +/- 148 ng/ml were achieved. ACTH and cortisol secretory patterns over a 24-hour period were also investigated in one subject during each trimester of pregnancy. Diurnal variations were observed that were qualitatively similar to those seen in nonpregnant women. From the results of these studies, we conclude that ACTH levels are suppressed in plasma of normal pregnant women but are higher in late pregnancy than in early pregnancy. The rise in plasma ACTH concentrations, as pregnancy advances, in spite of increasing levels of plasma cortisol, estrogens, and progesterone, is suggestive of the possibility that a source of ACTH exists that is not subject to negative feedback control, that the clearance of free cortisol increases as pregnancy advances, or that there is an alteration in the metabolism of the ACTH precursor protein produced by the pituitary and/or placenta.
A model proposed for regulation of steroidogenesis, lipoprotein utilization and cholesterol metabolism in HFA tissue is presented in Fig 17. We envision that the role of ACTH and cAMP in steroidogenesis and cholesterol metabolism is as follows. ACTH binds to specific receptors on the surface of the cells of the HFA gland and as a consequence, adenylate cyclase is activated, leading to increased formation of cAMP. cAMP causes activation of protein kinase that leads, presumably, to phosphorylation of specific proteins. This leads to the initiation of reactions that give rise to increased activity of key enzymes and levels of proteins involved in adrenal cholesterol metabolism. Presumably, the action of ACTH causes an increase in the activity of cholesterol side chain cleavage, the rate-limiting step in the conversion of cholesterol to steroid hormones. We suggest that once the mitochondrial cholesterol side-chain cleavage system is fully activated by ACTH, the supply of cholesterol to the mitochondria becomes rate-limiting for steroidogenesis. To meet this demand for cholesterol, a further action of ACTH results in an increase in the number of LDL receptors. LDL binds to specific receptors on the cell surface that are localized in coated pits. LDL is internalized by a process of adsorptive endocytosis and the internalized vesicles fuse with lysosomes and the protein component of LDL is hydrolyzed by lysosomal proteolytic enzymes to amino acids. The cholesteryl esters of LDL also are hydrolyzed to give rise to fatty acids and cholesterol. The liberated cholesterol is available for utilization in the biosynthesis of steroid hormones and other cellular processes. In addition, ACTH stimulates the activity of HMG CoA reductase and, thus, the rate of de novo cholesterol biosynthesis. In this way sufficient cholesterol is obtained to provide for precursor cholesterol to maintain the high rate of steroid synthesis by the HFA. HDL is not utilized as a source of cholesterol by the HFA. Because of the rapid rate of utilization of LDL by the HFA, fetal plasma levels of LDL are low and the activity of the HFA is a primary determinant of these levels. Thus, in the case of anencephaly, in which the activity of the adrenal is very low, plasma levels of LDL are 2--3 times higher than in normal fetuses, whereas plasma HDL levels are similar. In addition, in the normal neonate plasma LDL levels rise rapidly after birth, and this event is coincident with the involution of the fetal zone of the adrenal. The fetal liver is likely to be the major source ultimately of the LDL-cholesterol utilized by the HFA. Consequently, factors that regulate cholesterol and lipoprotein synthesis in the fetal liver may, in turn, affect the steroidogenic activity of the HFA through regulation of the supply of cholesterol precursor. Thus, if trophic factors for the HFA other than ACTH exist, an important site of their action might be the fetal liver, rather than a direct action to influence the rate of synthesis of steroids by the fetal adrenal.
In the present investigation, the mechanism(s) whereby ACTH stimulates low density lipoprotein (LDL) metabolism in human fetal adrenal was evaluated. Plasma membrane fractions were prepared from fetal adrenal tissue fragments incubated in lipoprotein-poor serum with or without ACTH, and the binding of [125I]iodo-LDL to such membrane fractions was examined. The mean specific binding capacity for [125I]iodo-LDL by membrane fractions prepared from four separate fresh human fetal adrenal glands was 1370 +/- 168 ng mg-1 protein (mean +/- SE), and the concentration of [125I]iodo-LDL producing half-maximal binding was 20.8 +/- 1.2 ng ml-1. Thus, the presence of high affinity, low capacity binding sites for LDL in human fetal adrenal tissue was confirmed. When human fetal adrenal tissue was maintained in organ culture for 2 days in medium containing lipoprotein-poor serum in the absence of ACTH, and plasma membrane fractions were subsequently prepared, the binding capacity for LDL in such membrane fractions was the same as or slightly greater than that in membrane fractions prepared from fresh tissue. When ACTH was present in the culture medium, the binding capacity for LDL was doubled compared to that in membrane fractions prepared from tissues incubated in the absence of ACTH. The rate of [125I]iodo-LDL degradation by human fetal adrenal tissue maintained in medium containing ACTH was also twice that of tissue maintained in the absence of ACTH. These results demonstrate that ACTH causes an increase in the number of LDL-binding sites in human fetal adrenal tissue in vitro. This is one mechanism whereby ACTH stimulates LDL metabolism in this tissue.
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