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![Reverse transcription-polymerase chain reaction (RT-PCR) analysis of total RNA from 72 h primary cytotrophoblast cultures (lanes 1 – 7), liver and kidney biopsies [11 β -hydroxysteroid dehydrogenase (11 β -HSD)-1 and -2-positive tissues respectively]. Bands correspond to the products of PCR using oligonucleotide primers speci fi c for ( A ) 11 β -HSD1. ( B ) 11 β -HSD2. ( C ) Mineralocorticoid receptor (MR). ( D ) Glucocorticoid receptor (GR).](profile/Mark-Kilby/publication/12054800/figure/fig5/AS:277025706266627@1443059483129/Reverse-transcription-polymerase-chain-reaction-RT-PCR-analysis-of-total-RNA-from-72-h.png)
Reverse transcription-polymerase chain reaction (RT-PCR) analysis of total RNA from 72 h primary cytotrophoblast cultures (lanes 1 – 7), liver and kidney biopsies [11 β -hydroxysteroid dehydrogenase (11 β -HSD)-1 and -2-positive tissues respectively]. Bands correspond to the products of PCR using oligonucleotide primers speci fi c for ( A ) 11 β -HSD1. ( B ) 11 β -HSD2. ( C ) Mineralocorticoid receptor (MR). ( D ) Glucocorticoid receptor (GR).
Source publication
Interconversion of active and inactive glucocorticoids, e.g. cortisol (F) and cortisone (E) is catalysed by 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) which exists as two isoforms. We have used human placental bed biopsies and an in-vitro cytotrophoblast cell culture system to examine the expression and activity of the 11 beta-HSD isoforms...
Contexts in source publication
Context 1
... analyses of mRNA isolated from primary cultures of cytotrophoblasts demonstrated the presence of transcripts for both 11β-HSD1 and 11β-HSD2, with products corresponding to the predicted sizes of 451 and 773 kb respectively ( Figure 5A,B). Further analyses showed expression of both GR and MR, with RT-PCR products at 693 and 450 bp respectively ( Figure 5C,D). ...
Context 2
... analyses of mRNA isolated from primary cultures of cytotrophoblasts demonstrated the presence of transcripts for both 11β-HSD1 and 11β-HSD2, with products corresponding to the predicted sizes of 451 and 773 kb respectively ( Figure 5A,B). Further analyses showed expression of both GR and MR, with RT-PCR products at 693 and 450 bp respectively ( Figure 5C,D). ...
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Intrauterine growth restriction (IUGR) is associated with systemic hypertension of the offspring later in life. The exact mechanisms are still incompletely understood. 11β-Hydroxysteroid dehydrogenase 2 (11β-HSD2) in the distal renal tubule protects the mineralocorticoid receptor from cortisol. As we did not find a suppression of 11β-HSD2 in total...
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Citations
... Fetal exposure to excessive glucocorticoid levels has been associated with poor fetal development, resulting not only in intrauterine growth retardation, but also in increased risk for diseases later in life [177]. The placenta itself responds to circulating glucocorticoids via the glucocorticoid and mineralocorticoid receptor [178]. During unchallenged pregnancies with physiological maternal glucocorticoid levels, term female placentae exhibit higher GR expression and 11β-HSD2 activity compared to male ones [179,180]. ...
The intrauterine environment is an important determinant of immunity later in life of the offspring. An altered prenatal immune development can result in a high postnatal risk for infections, chronic immune diseases, and autoimmunity. Many of these immune diseases show a strong sex bias, such as a high incidence of autoimmune diseases and allergies in adult females or a high risk for infections in males. Here, we comprehensively review established pathways and propose novel concepts modulating the risk for such poor immunity during childhood and throughout life. Moreover, we highlight how an adverse fetal environment may affect or aggravate the risk for poor immunity in a sex-specific manner. An improved understanding of a sex-specific susceptibility to poor immunity along with insights on how such risk can be modulated before or around birth will allow the development of tailored prevention strategies.
... Maternal GCs have crucial roles in fetal growth and development during pregnancy, but excessive GCs in fetal circulation will induce abnormal fetal development (such as IUGR) and program individuals for diseases later in adult life (24). The placenta serves as an important barrier between maternal and fetal GCs, protecting the fetus from overexposure to GCs (25). Our previous studies have confirmed that prenatal nicotine exposure (PNE) can weaken the placental GC barrier, induce IUGR through fetal overexposure to maternal GCs (26), and increase blood TCH levels in adults (27). ...
Epidemiologic studies showed that low birth weight is associated with high cholesterol and an increased risk of cardiovascular diseases in adulthood. This study aimed to elucidate the intrauterine programming mechanisms of adult hypercholesterolemia. The results showed that prenatal nicotine exposure (PNE) caused intrauterine growth retardation and hypercholesterolemia in male adult offspring rats. Hepatic cholesterol synthesis and output were deceased in utero but increased in adults; hepatic reverse cholesterol transport (RCT) persistently deceased before and after birth. Meanwhile, PNE elevated serum corticosterone level and decreased hepatic IGF1 pathway activity in male fetuses, whereas converse changes were observed in male adults. The chronic stress model and cortisol‐treated HepG2 cells verified that excessive glucocorticoid (GC)‐induced GC‐IGF1 axis programming enhanced hepatic cholesterol synthesis and output. In addition, PNE decreased the expression of specific protein 1 and P300 enrichment and H3K27 acetylation at the promoter region of genes responsible for RCT both in fetal and adult, male livers and reduced expression of those genes, similar alterations were also confirmed in cortisol‐treated HepG2 cells, suggesting that excessive GC‐related programming induced continuous RCT reduction by epigenetic modification. Taken together, the “2‐programming” approach discussed above may ultimately contribute to the development of hypercholesterolemia in male adult offspring.—Zhou, J., Zhu, C., Luo, H., Shen, L., Gong, J., Wu, Y., Magdalou, J., Chen, L., Guo, Y., Wang, H. Two intrauterine programming mechanisms of adult hypercholesterolemia induced by prenatal nicotine exposure in male offspring rats. FASEB J. 33, 1110–1123 (2019). www.fasebj.org
... 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2) is encoded by the HSD11B2 gene, it converts potent 11β-hydroxyglucocorticoids (cortisol, corticosterone) into their inactive 11-keto forms (cortisone, 11-dehydrocorticosterone); it is expressed in the cytotrophoblast and syncytiotrophoblast in placenta and provides a barrier between maternal and fetal GCs (Driver et al., 2001). Over-exposure of the fetus to maternal GCs due to impaired 11β-HSD2 function can lead to low birthweight, which is recognized as the main mechanism of increased risk for metabolic diseases in later life (Stroud et al., 2016). ...
... 3.1.2. Effects of PNE on placental 11β-HSD2 expression, fetal serum/ placental corticosterone levels and placental/fetal weight Placental 11β-HSD2 is an important "barrier" between maternal and fetal serum GCs (Driver et al., 2001). We found that both the mRNA (P < 0.01, Fig. 2A) and the protein (P < 0.01, P < 0.05, Fig. 2B) levels of 11β-HSD2 were significantly decreased. ...
... In this study, we found that the expression of 11β-HSD2 is reduced both in PNE placentas and nicotine treated BeWo cells. 11β-HSD2 is not only a GCs barrier that plays an important protective role in the prevention of fetal exposure to maternal GCs (Driver et al., 2001), but it is also important for placental structure and function (Kosicka et al., 2017). We found that the mRNA expression of 11β-HSD2 in placenta was positively correlated with placental weight and fetal bodyweight (only in females) in this study. ...
... GR levels have been quantified in only a few reports. They have been reported for example as 893 fmol GR/ mg protein in healthy skin, 2777 fmol GR/mg protein in skin from AIDS patients (Guo et al., 1996), 16,200 fmol GR/cell in normal epithelial stem cells (Driver et al., 2001) and 81,000 molecules GR/cell in uterine cervical cancer cells (Lu et al., 2005). Thus where levels of GR shown to result in GC-independent GR activation have been quantified, these appear to be within the physiologically relevant range (Robertson et al., 2013), arguing that these effects are likely to occur in vivo. ...
... Human placental cells are GC-responsive cells expressing GRs and mineralocorticoid receptors (MRs; Driver et al. 2001Driver et al. , 2003Chan et al. 2003;Patel et al. 2003;Lee et al. 2005). Although GRs are expressed in most fetal tissues, fetal membranes and in the human placenta from the early embryonic stages (Cole et al. 1995b), the expression of MRs is more limited and present only during the later stages of fetal development, at least in animal models (Brown et al. 1996). ...
Glucocorticoids (GCs) control essential metabolic processes in virtually every cell in the body and play a vital role in the development of fetal tissues and organ systems. The biological actions of GCs are mediated via glucocorticoid receptors (GRs), the cytoplasmic transcription factors that regulate the transcription of genes involved in placental and fetal growth and development. Several experimental studies have demonstrated that fetal exposure to high maternal GC levels early in gestation is associated with adverse fetal outcomes, including low birthweight, intrauterine growth restriction and anatomical and structural abnormalities that may increase the risk of cardiovascular, metabolic and neuroendocrine disorders in adulthood. The response of the fetus to GCs is dependent on gender, with female fetuses becoming hypersensitive to changes in GC levels whereas male fetuses develop GC resistance in the environment of high maternal GCs. In this paper we review GR function and the physiological and pathological effects of GCs on fetal development. We propose that GC-induced changes in the placental structure and function, including alterations in the expression of GR mRNA and protein levels, may play role in inhibiting in utero fetal growth.
... 93 However, although the mineralocorticoid receptor is expressed in many tissues, such as the kidney, colon, heart, hippocampal formation, brown adipose tissue, and sweat glands, its expression is often protected from glucocorticoid activation through colocalization of HSD11K (11-betadehydrogenase isozyme 2), a GR-regulated gene, whose product inactivates cortisol to cortisone. [94][95][96][97][98] In the context of inflammatory disease, GR acts in a reciprocal manner with NFKB1, a phenomenon that has been well-characterized; although a genome-wide coactivation analysis shows that GRE and NFKB1 bind only in a fraction of GR-and p65-(NF-jb protein 65) binding sites. 78 Studies of neuronal PC-12 cells suggest that greater than 40% of GR-binding sites are not GREs, as defined by the canonical motif, and related motifs, consisting of A/G-G-ACA-T/A-GT-CTC. 87 Of the 1031 sites that bound the GR, only 25 were shared with 3857 GR-binding sites in A549 cells derived from human lung epithelium, and 73 shared with the 8265 GR-binding sites found in the mouse adipocyte 3T3-L1 cell line. ...
Background
Chronic psychological stress is associated with enhanced abdominal pain and altered intestinal barrier function that may result from a perturbation in the hypothalamic-pituitary-adrenal (HPA) axis. The glucocorticoid receptor (GR) exploits diverse mechanisms to activate or suppress congeneric gene expression, with regulatory variation associated with stress-related disorders in psychiatry and gastroenterology. PurposeDuring acute and chronic stress, corticotropin-releasing hormone drives secretion of adrenocorticotropic hormone from the pituitary, ultimately leading to the release of cortisol (human) and corticosterone (rodent) from the adrenal glands. Cortisol binds with the GR in the cytosol, translocates to the nucleus, and activates the NR3C1 (nuclear receptor subfamily 3, group C, member 1 [GR]) gene. This review focuses on the rapidly developing observations that cortisol is responsible for driving circadian and ultradian bursts of transcriptional activity in the CLOCK (clock circadian regulator) and PER (period circadian clock 1) gene families, and this rhythm is disrupted in major depressive disorder, bipolar disorder, and stress-related gastrointestinal and immune disorders. Glucocorticoid receptor regulates different sets of transcripts in a tissue-specific manner, through pulsatile waves of gene expression that includes occupancy of glucocorticoid response elements located within constitutively open spatial domains in chromatin. Emerging evidence supports a potentially pivotal role for epigenetic regulation of how GR interacts with other chromatin regulators to control the expression of its target genes. Dysregulation of the central and peripheral GR regulome has potentially significant consequences for stress-related disorders affecting the brain-gut axis.
... The maximum amount of endogenous GR in COS-1 is 61 nM based on the previously reported 111 000 molecules per cell (1420 fmol/mg) [17] and our finding of a 3 pL cell volume. Moreover, 16 200 fmol/mg in cytotrophoblasts [18] was calculated to be 701 nM using the same method. The range above a total EGFP-GR concentration of 50 nM in the microwell was ignored in this fitting model because the 50 nM total concentration of EGFP-GR corresponded to a 1.5 lM concentration in the cell calculated using the 3 pL cell volume, and a concentration of over 1.5 lM in a cell seemed to be overexpression compared with the endogenous concentration of GR. ...
Glucocorticoid receptor α (GR) binds to the promoter regions of target genes as a homodimer and activates its transcriptional process. Though the homodimerization is thought to be the initial and essential process, the dissociation constant for homodimerization of GR remains controversial. To quantify homodimerization of (enhanced green fluorescence protein) EGFP-(glucocorticoid receptor) GR, the particle brightness in lysates from single cell was estimated for the fraction of homodimeric EGFP-GR using fluorescence correlation spectroscopy and microwells. Fitting the data with a bimolecular reaction model, the dissociation constant was determined. Moreover slow-diffusion complex was observed. These results suggest that EGFP-GR forms not only a monomer-dimer equivalent state but also a large-molecular-weight complex.
Copyright © 2015. Published by Elsevier B.V.
... During pregnancy the maternal HPA-axis activity gradually increases overtime toward a state of hypercortisolism by steadily increasing production of corticotropin-releasing hormone (CRH) in the placenta and fetal membranes [9,51]. During this time, most of the maternal cortisol (80-90%) permeating the placental unit is converted to inactive cortisone by the enzyme, 11Bhydroxysteroiddehydrogenase type 2 (11B-HSD2), which is synthesized together with glucocorticoid receptors in trophoblastic cells of the fetoplacental unit and is crucial to protect the fetus from excess glucocorticoid exposure [52]. Thus, considering the extremely high maternal glucocorticoid levels compared to those of the fetus, and the increase in placental 11β-HSD-2 activity that occurs throughout gestation, subtle changes in 11β-HSD-2 activity during pregnancy may induce dramatic glucocorticoid effects on the fetus due to glucocorticoid toxicity [52]. ...
... During this time, most of the maternal cortisol (80-90%) permeating the placental unit is converted to inactive cortisone by the enzyme, 11Bhydroxysteroiddehydrogenase type 2 (11B-HSD2), which is synthesized together with glucocorticoid receptors in trophoblastic cells of the fetoplacental unit and is crucial to protect the fetus from excess glucocorticoid exposure [52]. Thus, considering the extremely high maternal glucocorticoid levels compared to those of the fetus, and the increase in placental 11β-HSD-2 activity that occurs throughout gestation, subtle changes in 11β-HSD-2 activity during pregnancy may induce dramatic glucocorticoid effects on the fetus due to glucocorticoid toxicity [52]. ...
Abnormal function of the hypothalamic–pituitary–adrenal (HPA) axis along with the sympathetic-adrenalmedullary (SAM) axis represents the hallmark pathological findings in non-pregnant and pregnant women exhibiting major depressive disorder (MDD). Perinatal depression is highly prevalent, with an estimated prevalence exceeding 10% in most high-income countries. Pregnant women experience more stressful life events than non-pregnant women. Pregnant women with MDD show higher baseline levels of cortisol, increased levels of proinflammatory cytokines, and higher levels of hypothalamic-pituitary peptide hormones, catecholamines, and low dehydroepiandrosterone (DHEA) levels in plasma compared to those without MDD. During pregnancy the innate immune system and placental factors play crucial roles in the development of the conceptus to its full term. These factors when altered may generate a persistent dysfunction of the HPA axis that could lead to an overt transfer of cortisol and toxicity to the fetus at the expense of reduced activity of placental 11β-hydroxysteroid dehydrogenase Type 2 (11β-HSD-2). Epigenetic modifications (DNA methylation) of gene enhancers of the glucocorticoid receptor (GR) in addition to the corticotrophin-releasing hormone (CRH) and arginine-vasopressin (AVP) among other brain and placental molecules, may significantly contribute to the expression of depression during pregnancy. Pregnant women exhibiting affective disorders have a higher risk of generating preterm babies with high rates of morbidity and mortality and negative outcomes in child development compared to pregnant women without these disorders. Affective disorders in pregnant women should be taken seriously, and therapies focused in reducing or at most preventing the deleterious effects induced by stressors should be implemented to promote the welfare of both mother and baby. The goal of this paper is to describe the modulatory role of distinct paracrine and endocrine mediators including epigenetics of hormone receptors that drive the abnormal activity of the HPA axis in perinatal depression.
... 11β-HSD1 in proximal cells and 11β-HSD2 in collecting duct cells of the kidney where both appear to function as a dehydrogenase [247]. Both are expressed and tightly regulated throughout gestation in the placenta, 11 beta-HSD1 in decidualized stromal cells on the maternal side of the placenta, 11 beta-HSD2 in villous cytotrophoblast, syncytiotrophoblasts and trophoblast cells from the fetus that invade the placental bed and maternal vasculature [248]. While most have found 11β-HSD1 expression in smooth muscle cells and 11β-HSD2 in endothelial cells of vessels [249], both enzymes were reported to be expressed in human aortic vascular and bronchial smooth muscle cells [250] 11β-HSD1, but not 11β-HSD2, in these smooth muscle cells was increased in by inflammatory cytokines, resulting in an increased net conversion of 3 H-cortisone to 3 H-cortisol [250]. ...
... Ambos os tipos tem sido demonstrados na placenta. No entanto, o tipo 11beta-HSD2 tem sido localizado na camada do sinciciotrofoblasto da placenta, nas trocas materno-fetal (Driver et al. 2001). De acordo com Van-Beek et al. (2004), o tratamento com dexametasona estimula a expressão da enzima 11beta-HSD2 e a atividade desta em culturas de células trofoblásticas da placenta. ...
The dexamethasone, a synthetic glucocorticoid, has the ability to cross the placenta by increasing the level of movement of corticosteroids from mother to fetus during pregnancy. When administered in the late stages of pregnancy can produce effects undesirable on placental formation. The present study aimed to investigate the effect of administration of dexamethasone (0.8mg/day/animal) in the first five days of pregnancy, on placental development in rats. We used 30 albino rats, divided into two groups: Group I -pregnant rats without the application of dexamethasone, sacrificed to the 7th and 14th day. Group II -rats subjected to the application of dexamethasone in the first five days of pregnancy, sacrificed to the 7th and 14th day. The results showed that dexamethasone did not affect the number and histology of the implantation sites, but promoted changes in the disk placental causing hypertrophy in trophoblastic giant cell layer. No changes were found in the content of collagen, but there was interference with the metabolism of glycogen in spongiotrophoblast. The morphometry of lines showed, a difference between groups in the region of labyrinth and trophoblast giant cell. However, in morphometry of points there was a difference between groups in the region of labyrinth.
