Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “TROPHOBLAST”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 253 records · Page 14Linked to original sources

Trophoblastic invasion and the development of uteroplacental arteries in the macaque: immunohistochemical localization of cytokeratins, desmin, type IV collagen, laminin, and fibronectin.

The processes by which trophoblast cells invade and modify the walls of the uteroplacental arteries of macaques during the course of gestation were examined. Antibodies to cytokeratins were employed to identify trophoblast, anti-desmin antibody to identify smooth muscle, and antibodies to type IV collagen, laminin, and fibronectin to examine changes in extracellular matrix distribution in the arterial wall. During early gestation, endovascular trophoblast adhered to the arterial wall, often in an asymmetrical distribution. As trophoblast cells moved outwardly into the tunica media, the basement membrane underlying the endothelium was lost, as indicated by gaps in the layer when stained for type IV collagen and laminin. Trophoblast cells became sequestered in the vessel wall where they hypertrophied and became surrounded by a capsule containing type IV collagen and laminin. As the trophoblast cells became established in the vessel wall, the muscular layer of the artery became discontinuous. Throughout gestation it was common for trophoblast cells to invade the vessel intimal layer and share the lining of the artery with typical endothelial cells. This general disposition of endovascular and intramural trophoblast persisted into late gestation. In addition, and contrary to the results of earlier studies of macaques, we identified trophoblastic invasion and modification of myometrial segments of the uteroplacental arteries in later gestation. We also found evidence of interstitial trophoblast cells among the stromal cells of the endometrium, especially during early gestation.

Animals↗

An ultrastructural study of interstitial implantation in captive-bred, short-tailed fruit bats, Carollia perspicillata: trophoblastic adhesion and penetration of the uterine epithelium.

Trophoblastic adhesion to, and penetration of, the uterine epithelium during implantation have been examined in captive-bred Carollia perspicillata at the light and electron microscopic levels. Initial adhesion is localized to marginal ridges immediately over the apical intercellular junctions of the epithelial cells. Penetration then involves the intrusion of trophoblastic processes between the epithelial cells and the formation of junctional complexes between the two cell types. As larger areas of adhesion develop, they still occur most often near the intercellular boundaries between the more flattened epithelial cells or on their lateral sides. This suggests that many (if not all) of these broad areas of adhesion to the trophoblast had actually formed along what had originally been part of the lateral (rather than the apical) surfaces of the epithelial cells. Portions of the apical cell surfaces further removed from their intercellular boundaries usually were not adherent to the trophoblast and still had microvilli. Upon reaching the basal lamina of the uterine epithelium, invasion of the endometrium is temporarily retarded, and trophoblast cells migrate instead between the basal lamina and the epithelial cells. This occurs extensively along both the luminal and glandular epithelia around the implantation site, but not significantly along their apical surfaces. This again indicates that adhesive interactions between the trophoblast and those apical surfaces are at most very limited in Carollia. The epithelial cells appear to be viable until separated from their basal laminae by the trophoblast. They are then phagocytized by the trophoblast. During initial penetration of the uterine luminal epithelium, the trophoblast is still entirely of the cellular variety. Syncytiotrophoblast does not begin to appear until later, when the trophoblast first comes into contact with endometrial capillaries.

Animals↗

Murine trophoblast resists cell-mediated lysis. II. Resistance to natural cell-mediated cytotoxicity.

The susceptibility of murine trophoblast cells to natural cell-mediated cytotoxicity has been assessed. Primary short-term cultures of murine trophoblast cells isolated from 14-day placentas were found to be resistant to endogenous and interferon-activated natural killer (NK) cells and natural cytotoxic cells. That the relevant target structures are expressed on the surface of trophoblast cells and accessible to the effectors was demonstrated by their ability to inhibit the lysis of NK-sensitive target cells (YAC-1) in a dose-dependent manner. The lytic resistance of trophoblast cells was unaffected by neuraminidase treatment, inhibition of protein synthesis, or extending the assay time to 12 hr. Moreover, trophoblast cells were resistant to antibody-dependent cell-mediated cytotoxicity when coated with an alloantibody capable of mediating their lysis in the presence of heterologous complement. Neither the preincubation of effector cells in concentrated trophoblast culture supernatants nor the direct exposure of effectors to monolayers of trophoblast cells inhibited their NK lytic activity, indicating that the secretion of a suppressive factor or the direct inactivation of the NK cells was not responsible for the observed resistance to lysis. These observations, together with previous results showing the resistance of trophoblast to cytotoxic T cell-mediated lysis, reveal that murine trophoblast cells possess a resistance mechanism against several forms of cell-mediated lysis. This feature of trophoblast cells at the maternal-fetal interface is likely to play an important role in protecting the fetoplacental allograft from immune rejection.

Animals↗

Trophoblast giant cell release of placental lactogens: temporal and regional characteristics.

Placental lactogen (PL) production by rat trophoblast giant cells was studied using in vitro methods. The influence of trophoblast giant cell location within the conceptus and day of trophoblast giant cell isolation on the type of PL released in vitro were investigated. The effect of trophoblast giant cell location on the amount of PL, progesterone, and testosterone released in vitro was also evaluated. Trophoblast giant cells release two types of PLs in vitro; a high-molecular-weight lactogen, PL-1, and a low-molecular-weight lactogen, PL-2. The type of PL released by trophoblast giant cells was not influenced by their location within the conceptus at the time of dissection. Location did influence the amount of hormone produced by trophoblast giant cells. Mural trophoblast giant cells were more active in the production of PL, progesterone, and testosterone. The type of PL released by trophoblast giant cells is highly dependent upon the day of gestation the cells are removed for study. Trophoblast giant cells isolated on Day 10 of gestation release predominantly PL-1, while those cells isolated 24 hr later (Day 11 of gestation) release predominantly PL-2. The switch from PL-1 to PL-2 production that occurs in vivo does not occur under the in vitro conditions employed in this report.

Animals↗

GM-CSF and CSF-1 stimulate DNA synthesis but not cell proliferation in short-term cultures of mid-gestation murine trophoblast.

Short-term cultures of purified murine trophoblast were used to investigate the potential trophic effects of a number of cytokines. Both granulocyte-macrophage colony stimulating factor (GM-CSF) and colony stimulating factor-1 (CSF-1) increased [3H]thymidine (TdR) uptake (3-8 times control values) by trophoblast harvested from placentae on day 12 or 14 of pregnancy. In contrast, interleukin-3 (IL-3) had only a mild stimulatory effect ([3H]TdR uptake 1.5 times control), and IL-2 did not alter the level of DNA synthesis in these cells. Immunocytochemical analysis confirmed that the cells engaged in DNA synthesis were cytokeratin-positive trophoblast cells and revealed that these cells predominantly bore markers (alkaline phosphatase, transferrin receptors) characteristic of trophoblast cells from the placental labyrinth. The increased DNA synthesis observed after exposure to GM-CSF or CSF-1 was not associated with a change in the proportion of nuclei involved in synthesis, nor did it result in significantly increased trophoblast cell numbers in the cultures. These findings suggest that DNA-synthesizing trophoblast cells were not proliferating, but were more likely engaged in endoreduplicative cycles leading to the formation of terminally differentiated trophoblast giant cells. These results caution against the presumption of proliferation when measuring [3H]thymidine incorporation by placental or trophoblast cells in standard in vitro cultures. In addition, taken together with the reports of high levels of CSF-1 in the pregnant uterus and the expression of the CSF-1 receptor on placental trophoblast cells, they suggest that the hemopoietic cytokines may play a role in the differentiation and/or function of trophoblast cells in the developing murine placenta.

Animals↗

Expression of cFABP and PPAR in trophoblast cells: effect of PPAR ligands on linoleic acid uptake and differentiation.

Throughout gestation, fetal growth depends, in part, on placental transfer of maternal essential fatty acid (EFA) and long-chain polyunsaturated fatty acid. All fatty acid (FA) can cross lipid bilayer by simple diffusion, such as those in the syncytiotrophoblasts, the multinucleated, terminally differentiated trophoblast cells. The trophoblasts differentiation process is accompanied by an increase of human chorionic gonadotropin (hCG) secretion and an inhibition of Human Achaete-Scute Homologue-2 expression (Hash-2). Furthermore, a number of FA-binding proteins (FABPs) have been identified in membrane and cytoplasm of mammalian cells, which are thought to facilitate the transfer of FA across membranes and their intracellular channeling. Thus, the aim of this study was to investigate the implication of cFABPs in linoleic acid (LA) uptake by human trophoblast cells according to differentiation. Moreover, since peroxisome proliferator-activated receptor (PPARs) regulate the expression of cFABP and play an important role in trophoblast cells differentiation, the effects of PPARs ligands are verified on cFABP expression and differentiation. Herein, we reported the increase of the expression of liver and heart FABP (L- and H-FABP) upon differentiation of trophoblast cells, an inhibition of PPAR alpha and beta, while PPAR gamma levels remains unchanged. The nonselective peroxisome-proliferating agents, bezafibrate and LA, impaired trophoblast differentiation, and reduced L- and H-FABP expression. Furthermore, cobalt, a chemical agent known to mimic hypoxia, inhibits trophoblast cells differentiation and diminishes H-, L-FABP and PPARs expression. Finally, both treatments show no influence on LA uptake by trophoblast cells. In conclusion, this study showed that there is no correlation between the expression of H- and L-FABP and LA uptake by trophoblast cells and that bezafibrate and LA greatly impaired trophoblast cells differentiation.

Basic Helix-Loop-Helix Proteins↗

ICAM-1 is overexpressed by villous trophoblasts in placentitis.

Although an in vitro study has hypothesized that expression of ICAM-1 by villous trophoblasts could be important for the influx of maternal immune cells in villitis, it remains to be shown whether the same phenomenon occurs in human villitis. To investigate the expression of ICAM-1 by villous trophoblasts, its relationship with rupture of the trophoblastic barrier and influx of immune cells into the villi, we analysed 18 paraffin-embedded placentas with placentitis (5 by Toxoplasma gondii, 3 by Trypanosoma cruzi, 2 by Paracoccidioides brasiliensis and 8 of unknown aetiology - VUE) and 8 control placentas for detection of ICAM-1 by immunohistochemistry. All cases but one of placentitis showed trophoblast overexpression of ICAM-1 in the inflamed villi, located almost exclusively next to the areas of trophoblastic rupture. The villitis cases (caused by T. cruzi, T. gondii and VUE) presented leukocyte adherence in the areas of trophoblastic rupture. When the inflammatory reaction was situated in the intervillous space (placentitis by P. brasiliensis), in spite of the trophoblastic rupture and ICAM-1 overexpression there was no leukocyte influx into villi. None of the control placentas showed ICAM-1 expression by the trophoblast. We concluded that overexpression of ICAM-1 by villous trophoblasts occurs during placentitis characterized by accumulation of leukocytes in the villous or intervillous space and probably plays an important role in the rupture of the trophoblastic barrier. The influx of immune cells into the villi appears to be mediated by ICAM-1 but the location of the antigen within villous stroma is certainly a crucial factor for its occurrence.

Adult↗

Macaque trophoblast migration is regulated by RANTES.

In human and non-human primates, migratory trophoblasts penetrate the uterine epithelium, invade the endometrium, enter the uterine vasculature, and migrate within the arteries. The mechanisms that regulate this directional migration are unknown. We have used early gestation macaque trophoblasts to test the hypothesis that trophoblast migration is regulated by the chemokine, Regulated on Activation T-Cell Expressed and Secreted (RANTES). Immunohistochemical analysis of cryosections of endometrial tissue showed expression of RANTES by stromal cells and vascular cells. Isolated endothelial cells expressed RANTES as determined by immunocytochemistry and RT-PCR analyses. Immunohistochemical analysis of endometrial cryosections showed that the RANTES receptor, CCR5, was expressed by trophoblasts on anchoring villi and by cells within the trophoblastic shell. Cytokeratin-positive/CCR5-positive cells, consistent with trophoblasts, were also found scattered within the stroma and were often clustered around blood vessels. Isolated trophoblast cells expressed CCR5 as determined by immunocytochemistry and RT-PCR analyses. Isolated trophoblasts migrated towards RANTES when cultured in migration chambers and migration was reduced in the presence of anti-CCR5 antibody. When trophoblasts were cultured on dishes coated with recombinant RANTES, expression of beta1 integrin was increased. The RANTES-induced increase in beta1 integrin expression was inhibited by pertussis toxin. These data suggest a role for RANTES and CCR5 in the regulation of trophoblast migration within the endometrium and within the uterine vasculature.

Animals↗

Expression of maspin in gestational trophoblastic disease.

BACKGROUND: Maspin is a tumor suppressor gene whose expression is altered in neoplasia and malignancies of many tissues. In the human placenta, the maspin gene is expressed in trophoblastic cells and might act as an inhibitory regulator of trophoblastic invasion. Hence, in gestational trophoblastic disease (GTD), where there is increased propensity for invasion in the trophoblastic tissue, we hypothesized that maspin expression would be decreased. The present study aimed at investigating the expression of maspin in GTD and its prognostic significance. METHODS: Using immunohistochemical staining, we firstly studied the expression of maspin in hydatidiform moles, with gestational age-matched normal first trimester placenta used as control. A total of 38 cases of hydatidiform moles were studied, including 20 complete moles (CM) and 18 partial moles (PM). Among them, 10 cases of the CM group and 8 cases of the PM group subsequently developed gestational trophoblastic neoplasia (GTN). Immunostaining was also performed on tissue from 4 cases of choriocarcinoma and 5 cases of placental site trophoblastic tumor. Reverse transcriptase-polymerase chain reaction (RT-PCR) was further performed on RNA extracted from 10 hydatidiform moles (5 with GTN and 5 without) and 6 normal first-trimester placentae. RESULTS: In all tissue sections, nuclear expression of immunostaining signal was demonstrated, mainly in the cytotrophoblasts. The percentage of trophoblastic nuclei stained in both complete and partial moles was significantly lower than that in normal first-trimester placenta (P < 0.001). However, there was no significant difference in immunostaining between complete and partial moles (P > 0.05). There was also significantly lower expression of maspin in those cases subsequently developing GTN than those which did not (P = 0.01). Immunostaining on choriocarcinoma and placental site trophoblastic tumor showed reduced expression of maspin in all the tumor cells. Reverse transcriptase-polymerase chain reaction revealed that the expression of maspin was consistently down-regulated in all the hydatidiform mole samples. CONCLUSIONS: Our results suggest that there is down-regulated expression of maspin in gestational trophoblastic diseases, and the down-regulation is more prominent in cases developing gestational trophoblastic neoplasia. This may play a role with prognostic significance in the pathogenesis and malignant transformation of hydatidiform moles.

Choriocarcinoma↗

Evidence of embryo- and trophoblast-toxic cellular immune response(s) in women with recurrent spontaneous abortion.

OBJECTIVE: The purpose of this study was to determine whether reproductive antigens stimulate lymphocytes and macrophages from women with recurrent abortion to secrete factors that are toxic to preimplantation embryos or trophoblast cells in vitro. STUDY DESIGN: Mononuclear cells were isolated from 30 fertile controls and 300 nonpregnant women being evaluated for recurrent abortion. Supernatants generated from these cells after separate culture with sperm and trophoblast antigen extracts were added to two-cell mouse embryo cultures and trophoblast proliferation assays. Toxicity was assumed when the median percentage of embryos developing to blastocysts or trophoblast proliferation was less than or equal to 50% of control values. Both parametric and nonparametric statistical methods were used where appropriate. RESULTS: Mouse embryo development and/or trophoblast proliferation were significantly inhibited by supernatants from trophoblast and/or sperm antigen-activated peripheral blood leukocyte cultures from a majority of 300 women with recurrent abortion but not from 30 women with normal reproductive histories. The mouse blastocyst development assay was more sensitive than the trophoblast proliferation assay in determining toxic factor production. Embryo-toxic factors were produced by activated leukocyte cultures from 90% of 180 women with a history of recurrent abortion of unexplained etiology, whereas trophoblast-inhibitory factors were detected in 50% of women from the same group. The embryo-toxic factor(s) was heat labile, had a molecular weight(s) between 10 and 30 kd, and was absorbed out by passage through affinity columns containing anti-interferon gamma beads. CONCLUSION: We conclude that recurrent abortion in some women is associated with embryo- and/or trophoblast-toxic factor production in response to stimulation by sperm or trophoblast antigens and that the principal factor may involve the 18 kd, heat-labile, T-lymphocyte cytokine interferon gamma. This study suggests a new cause of recurrent abortion.

Abortion, Habitual↗

Interactions between trophoblast cells and the maternal and fetal circulation in the mouse placenta.

Mammalian embryos have an intimate relationship with their mothers, particularly with the placental vasculature from which embryos obtain nutrients essential for growth. It is an interesting vascular bed because maternal vessel number and diameter change dramatically during gestation and, in rodents and primates, the terminal blood space becomes lined by placental trophoblast cells rather than endothelial cells. Molecular genetic studies in mice aimed at identifying potential regulators of these processes have been hampered by lack of understanding of the anatomy of the vascular spaces in the placenta and the general nature of maternal-fetal vascular interactions. To address this problem, we examined the anatomy of the mouse placenta by preparing plastic vascular casts and serial histological sections of implantation sites from embryonic day (E) 10.5 to term. We found that each radial artery carrying maternal blood into the uterus branched into 5-10 dilated spiral arteries located within the metrial triangle, populated by uterine natural killer (uNK) cells, and the decidua basalis. The endothelial-lined spiral arteries converged together at the trophoblast giant cell layer and emptied into a few straight, trophoblast-lined "canals" that carried maternal blood to the base of the placenta. Maternal blood then percolated back through the intervillous space of the labyrinth toward the maternal side of the placenta in a direction that is countercurrent to the direction of the fetal capillary blood flow. Trophoblast cells were found invading the uterus in two patterns. Large cells that expressed the trophoblast giant cell-specific gene Plf (encoding Proliferin) invaded during the early postimplantation period in a pattern tightly associated with spiral arteries. These peri/endovascular trophoblast were detected only approximately 150-300 microm upstream of the main giant cell layer. A second type of widespread interstitial invasion in the decidua basalis by glycogen trophoblast cells was detected after E12.5. These cells did not express Plf, but rather expressed the spongiotrophoblast-specific gene Tpbp. Dilation of the spiral arteries was obvious between E10.5 and E14.5 and was associated with a lack of elastic lamina and smooth muscle cells. These features were apparent even in the metrial triangle, a site far away from the invading trophoblast cells. By contrast, the transition from endothelium-lined artery to trophoblast-lined (hemochorial) blood space was associated with trophoblast giant cells. Moreover, the shaping of the maternal blood spaces within the labyrinth was dependent on chorioallantoic morphogenesis and therefore disrupted in Gcm1 mutants. These studies provide important insights into how the fetoplacental unit interacts with the maternal intrauterine vascular system during pregnancy in mice.

Animals↗

Expression of HOX gene products in normal and abnormal trophoblastic tissue.

OBJECTIVE: The expression pattern of three homeobox genes products, HOX A11, HOX B6, and HOX C6, was examined in normal human placental tissue and abnormal trophoblastic tissue derived from complete hydatidiform moles and choriocarcinoma tumors. We sought to determine whether expression of these gene products during different states of trophoblastic differentiation and proliferation is constant or demonstrates variation. Variation in expression of these respective homeobox genes may provide insight into predicting which molar tissues are likely to develop into choriocarcinoma tumors. METHODS: Tissue sections from a total of 12 samples were studied. Among these, six full-term human placentas, three complete hydatidiform moles, and three choriocarcinoma tumors were examined for expression of the homeobox HOX A11, HOX B6, and HOX C6 gene products, using immunohistochemistry staining methods. RESULTS: Expression of HOX homeobox gene products, HOX A11, HOX B6, and HOX C6, was detected in full-term human placenta and tissue from complete hydatiform moles. Abnormal trophoblasts from complete moles demonstrated an immunoreactivity expression pattern comparable to that of normal trophoblasts from term pregnancies. However, definitive expression of these respective homeobox genes was not identified in tissue obtained from choriocarcinoma tumors. CONCLUSION: Variation in expression of HOX homeobox gene products, HOX A11, HOX B6, and HOX C6, was established in trophoblast tissue obtained from full-term human placentas, complete hydatiform moles, and choriocarcinoma tumors. This finding indicates that normal full-term trophoblasts and abnormal molar trophoblasts may share similar fundamental regulatory control mechanisms. The absence of definitive expression of these HOX gene products in trophoblastic cells derived from choriocarcinoma tumors indicates that while HOX A11, HOX B6, and HOX C6 genes may be involved in maintenance of some trophoblastic cell states, they may be either downregulated or have alterations in their expression in trophoblasts from choriocarcinoma tumors.

Adult↗

Vascular endothelial growth factor, placenta growth factor and their receptors in isolated human trophoblast.

The expression of the angiogenic growth factors, vascular endothelial cell growth factor (VEGF) and placenta growth factor (PIGF) was demonstrated in isolated human term cytotrophoblast and in vitro differentiated syncytiotrophoblast. RNase protection assays demonstrated VEGF expression in both cytotrophoblast and syncytiotrophoblast while prominent PIGF expression was detected in both types of trophoblast by Northern blot analyses. VEGF expression increased approximately eightfold in trophoblast cultured under hypoxic conditions (1 per cent O2) yet PIGF expression decreased 73 +/- 5.5 per cent in the same trophoblast. These results suggest distinct regulatory mechanisms govern expression of VEGF and PIGF in trophoblast. Characterization of the VEGF/PIGF receptors, KDR and flt-1, revealed the presence of flt-1 mRNA in isolated cytotrophoblast and in vitro differentiated syncytiotrophoblast. KDR was not detected in the isolated trophoblast. Exogenous rhVEGF induced c-Jun N-terminal kinase (JNK) activity in the normal trophoblast indicating that the flt-1 receptors on trophoblast are functional. Trophoblast-derived VEGF/PIGF could act in a paracrine fashion to promote uterine angiogenesis and vascular permeability within the placental bed. In addition, presence of function flt-1 on normal trophoblast suggests that VEGF/PIGF functions in an autocrine manner to perform an as yet undefined role in trophoblast invasion, differentiation, and/or metabolic activity during placentation.

Blotting, Northern↗

Quantitative analysis of trophoblast invasion in preeclampsia.

BACKGROUND: The process of physiological conversion of spiral arteries is dependent on the invasion of the interstitium and spiral arteries of the uterine wall by invasive extravillous trophoblast thereby creating a high flow-low resistance vessel. Quantitative data on restriction of trophoblast invasion and failure of spiral artery transformation are limited in preeclampsia. AIM: This study morphometrically analyzes interstitial trophoblast cells and trophoblast cells embedded in the wall of the converted spiral arteries within the decidua and myometrium of normotensive and preeclamptic Black African pregnant women. METHODS: Placental bed biopsies were obtained from 25 normotensive pregnant women and 30 pregnant women complicated with hypertensive disorders. Biopsies were processed and immunostained for trophoblast cell identification, using anti-MNF 116 antibody. Image analysis of the trophoblast population within the decidua, myometrium and the spiral arteries was performed in the normotensive and in the severe proteinuric hypertensive groups. RESULTS: The mean field area percentage of trophoblast cells observed in the decidua of the normotensive women was 22.79 +/- 2.1% in comparison to 18.14 +/- 1.53% in the severe hypertensive group (p < 0.01). In the myometrium, the mean field area percentage of interstitial trophoblast cells (both mononuclear and multinuclear) was 10.04 +/- 2.1% of the field area of the normotensive group compared to 2.81 +/- 0.67% in the severe hypertensive group (p < 0.001). The mean field area percentage occupied by trophoblast cells in myometrial spiral arteries was 10.15% in the normotensive group compared to none in the severe hypertensive group. The latter group displayed medial disorganization, hyperplasia and endothelial vacuolation. CONCLUSION: This study demonstrates restricted invasion of the trophoblastic cells in preeclampsia. This inadequate invasion may influence vascular remodeling required for delivering adequate volumes of maternal blood to the placenta.

Adult↗

Connexin expression patterns in human trophoblast cells during placental development.

This study focuses on the gap junction expression pattern in trophoblast cells during human placental development in vivo and in vitro. Investigations of cell-cell communication properties within the subpopulations of trophoblast responsible for invasion, placental growth and feto-maternal transport seem of special interest because the intercellular channels are believed to coordinate proliferation and differentiation processes. From all gap junction connexins (Cx) investigated (Cx26, Cx31, Cx32, Cx37, Cx40, Cx43), Cx40 was the only connexin clearly detected within the cytotrophoblast of human placenta, and was restricted to the extravillous trophoblast of cell islands and cell columns. Most intense staining was found in the juxtastromal area correlated to the proliferating extravillous trophoblast cells. Connexin protein expression was missing during trophoblast migration into the decidua but was re-expressed in trophoblast aggregates within the decidua. Cx40 expression decreased with progressing pregnancy and no connexins could be detected in villous or extravillous trophoblast of mature placentae. In parallel, isolated trophoblast cells of first and second trimester placentae revealed Cx40 expression and, in contrast to the situation in vivo, Cx43 was also found. In isolated cells of mature placentae, expression of both Cx40 and Cx43 transcripts was decreased to low levels and Cx40 immunoreactivity was absent. Cx43 protein, however, was still detectable in trophoblast cultures of term placentae. Our studies suggest that Cx40 is the characteristic channel for the proliferating cell population of cell islands and cell columns of first and second trimester placentae and isolated trophoblast and is probably involved in regulation and coordination of the invasive pathway.

Cell Division↗

Increased superoxide generation is associated with decreased superoxide dismutase activity and mRNA expression in placental trophoblast cells in pre-eclampsia.

Pre-eclampsia is a multi-system disorder unique to human pregnancy. Although the aetiology of pre-eclampsia is still unknown, increased placental oxidative stress contributes to the pathophysiology of this pregnancy disorder. The goal of this study was to determine if placental trophoblast cells generate superoxide, and if there was a difference in superoxide generation and superoxide dismutase (SOD) activity between trophoblast cells isolated from pre-eclamptic placentae versus normal placentae. Placentae were obtained from nine normal and 10 pre-eclamptic pregnancies immediately after delivery. Trophoblast cells were isolated and purified by Percoll density gradient centrifugation. Superoxide generation and SOD activity were determined by spectrophotometric assays. Localization of CuZn-SOD protein within the placenta was examined by immunohistochemical staining. mRNA expression of CuZn-SOD was determined in trophoblast cells isolated from five normal and five pre-eclamptic pregnancies by Northern blot analysis. 18S ribosomal mRNA expression was used as an internal standard. We found: (1) trophoblast cells from pre-eclamptic placentae generated significantly more superoxide than trophoblast cells from normal placentae: 17.62+/-3.19 versus 4.70+/-0.76 nmol/5x10(6) cells (mean+/-s.e.), P< 0.01; (2) protein and mRNA expression for CuZn-SOD was mainly localized in the trophoblast cells within the placenta; and (3) SOD activity and relative mRNA expression for CuZn-SOD were significantly decreased in trophoblast cells from pre-eclamptic placentae as compared to trophoblast cells from normal placentae, SOD activity: 6.46+/-1.76 versus 13.01+/-1.67 units/mg protein, P< 0.05; relative mRNA expression for CuZn-SOD: 0.25+/-0.09 versus 0.73+/-0.07, P< 0.01. We conclude that increased superoxide generation was associated with decreased SOD activity and mRNA expression for CuZn-SOD in trophoblast cells isolated from pre-eclamptic placentae. These findings support the notion of increased oxidative stress in the pre-eclamptic placenta, which may contribute to the pathophysiology of this pregnancy disorder.

Chorionic Gonadotropin, beta Subunit, Human↗

Characterization of a TGFbeta-responsive human trophoblast-derived cell line.

The placenta is formed by developing trophoblast cells to facilitate fluid, gas and nutrient exchange with the mother. Inappropriate trophoblast responsiveness can lead to life threatening complications during pregnancy including intrauterine growth retardation, pre-eclampsia, spontaneous abortion and malignancy that could lead to fetal loss. Transforming growth factor beta (TGFbeta) is a multifunctional cytokine required for embryonic development and is an important regulator of human trophoblast function. Although TGFbeta is critical for placental and embryonic development, there are currently no established TGFbeta-responsive human trophoblast-derived cell lines available to study the mechanisms by which TGFbeta regulates trophoblast function. Our studies have examined the transformed human trophoblast-derived cell line, ED27, to determine if it is responsive to TGFbeta. Our data indicate that TGFbeta dose responsively and reversibly inhibits cell growth in ED27 cells and induces classic TGFbeta response genes, fibronectin and plasminogen activator inhibitor 1 (PAI-1). TGFbeta also induces an inhibitor of trophoblast invasion, tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in ED27 cells. Our studies have identified a human trophoblast-derived cell line that parallels isolated primary human trophoblasts in their responses to TGFbeta. This cell line may provide us with the opportunity to determine TGFbeta-mediated responses on human trophoblast functions not previously possible.

Apoptosis↗

Macaque trophoblast migration toward RANTES is inhibited by cigarette smoke-conditioned medium.

Trophoblast migration within the endometrium and uterine vasculature is essential for normal placental and fetal development. We previously demonstrated that macaque trophoblasts express the chemokine receptor CCR5 and that this receptor mediates trophoblast migration toward RANTES (regulated upon activation normal T-cell expressed and secreted). In the present paper we have used primary cultures of early gestation macaque trophoblasts to test the hypothesis that tobacco smoke inhibits trophoblast migration as the result of dysregulation of the RANTES/CCR5 chemotactic axis. Early gestation macaque trophoblasts were incubated in the absence or presence of cigarette smoke-conditioned medium (CSM). Cell migration was quantified using migration chambers. CCR5 and G protein receptor kinase 2 (GRK2) expression was measured by immunofluorescence microscopy and Western blotting. cAMP levels were measured by enzyme-linked immunosorbent assay. Trophoblast migration toward RANTES was reduced when cells were incubated in CSM. Trophoblasts also showed reduced expression of CCR5, increased levels of cAMP, and increased expression of GRK2. Finally, the secretion of RANTES by uterine endothelial cells was reduced by exposing the cells to CSM. These results support the idea that cigarette smoke constituents inhibit directional trophoblast migration by causing increased desensitization of trophoblast CCR5 and inhibiting the secretion of RANTES by endothelial cells.

Animals↗