Feto-maternal Junction Research Articles

Quantitative assessment of morphology and sub-cellular changes in macrophages and trophoblasts during inflammation.

In pregnancy during an inflammatory condition, macrophages present at the feto-maternal junction release an increased amount of nitric oxide (NO) and pro-inflammatory cytokines such as TNF-α and INF-γ, which can disturb the trophoblast functions and pregnancy outcome. Measurement of the cellular and sub-cellular morphological modifications associated with inflammatory responses are important in order to quantify the extent of trophoblast dysfunction for clinical implication. With this motivation, we investigated morphological, cellular and sub-cellular changes in externally inflamed RAW264.7 (macrophage) and HTR-8/SVneo (trophoblast) using structured illumination microscopy (SIM) and quantitative phase microscopy (QPM). We monitored the production of NO, changes in cell membrane and mitochondrial structure of macrophages and trophoblasts when exposed to different concentrations of pro-inflammatory agents (LPS and TNF-α). In vitro NO production by LPS-induced macrophages increased 22-fold as compared to controls, whereas no significant NO production was seen after the TNF-α challenge. Under similar conditions as with macrophages, trophoblasts did not produce NO following either LPS or the TNF-α challenge. Super-resolution SIM imaging showed changes in the morphology of mitochondria and the plasma membrane in macrophages following the LPS challenge and in trophoblasts following the TNF-α challenge. Label-free QPM showed a decrease in the optical thickness of the LPS-challenged macrophages while TNF-α having no effect. The vice-versa is observed for the trophoblasts. We further exploited machine learning approaches on a QPM dataset to detect and to classify the inflammation with an accuracy of 99.9% for LPS-challenged macrophages and 98.3% for TNF-α-challenged trophoblasts. We believe that the multi-modal advanced microscopy methodologies coupled with machine learning approach could be a potential way for early detection of inflammation.

Spatiotemporal immunofluorescent evaluation of porcine reproductive and respiratory syndrome virus transmission across the maternal-fetal interface.

Porcine reproductive and respiratory syndrome virus (PRRSV) infection causes severe reproductive failure characterized by high fetal morbidity and mortality leading to substantial economic losses to the swine industry. Evaluation of spatiotemporal transmission of PRRSV at the maternal-fetal interface (MFI) is critical for understanding fetal infection. Localization of PRRSV-2 strain NVSL 97-7895 at different regions of the MFI in 20 pregnant gilts at 2, 5, 8, 12 and 14 days post-inoculation (dpi) were analyzed by immunofluorescence (IF). Samples of MFI were collected from 15 inoculated and 5 control gilts and transplacental PRRSV transmission assessed in randomly selected fetuses from each litter. Localization of NVSL 97-7895 antigen immunoreactivity in the MFI was focused in three major areas: endometrial connective tissues (ENDO), the feto-maternal junction (FMJ) and fetal placenta (PLC). NVSL 97-7895 was detected at the FMJ by 2 dpi. At 2, 5 and 8 dpi, NVSL 97-7895 was localized within the ENDO and FMJ, whereas at 12 and 14 dpi, it was mainly localized in the PLC. Using a novel IF strategy for counting and size sorting NVSL 97-7895 viral antigen in situ, results of this study indicate that non-cell-associated mechanisms are involved in PRRSV transmission across the MFI.

Identification of pregnancy-associated glycoproteins and alpha-fetoprotein in fallow deer (Dama dama) placenta

BackgroundThis paper describes the isolation and characterization of pregnancy-associated glycoproteins (PAG) from fetal cotyledonary tissue (FCT) and maternal caruncular tissue (MCT) collected from fallow deer (Dama dama) pregnant females. Proteins issued from FCT and MCT were submitted to affinity chromatographies by using Vicia villosa agarose (VVA) or anti-bovine PAG-2 (R#438) coupled to Sepharose 4B gel. Finally, they were characterized by SDS-PAGE and N-terminal microsequencing.ResultsFour distinct fallow deer PAG (fdPAG) sequences were identified and submitted to Swiss-Prot database. Comparison of fdPAG with PAG sequences identified in other ruminant species exhibited 64 to 83% identity. Additionally, alpha-fetoprotein was identified in fetal and maternal tissues.ConclusionOur results demonstrate the efficacy of VVA and bovine PAG-2 affinity chromatographies for the isolation of PAG molecules expressed in deer placenta. This is the first report giving four specific amino acid sequences of PAG isolated from feto-maternal junction (FCT and MCT) in the Cervidae family.

747: Hepatocyte growth factor (HGF) is upregulated at the fetomaternal junction in preeclamptic placentas

747: Hepatocyte growth factor (HGF) is upregulated at the fetomaternal junction in preeclamptic placentas

Distribution of CD14+ and CD68+ Macrophages in the Placental Bed and Basal Plate of Women With Preeclampsia and Preterm Labor

Objective Macrophages play a key role in implantation, placentation and parturition. Yet, whether or not the number of macrophages at the fetomaternal interface (basal plate of the placenta and placental bed) is altered in women with preeclampsia is the subject of controversy. The purpose of this study was to compare the immunoreactivity and distribution patterns of CD14 and CD68 positive macrophages in both the basal plate and placental bed from preeclamptic and non-preeclamptic pregnancies. Methods A cross-sectional study was conducted. Paraffin embedded sections of placental tissues and placental bed biopsies were obtained from patients with early onset preeclampsia ( n = 10) and from those with preterm labor/delivery ( n = 10) without preeclampsia matched for gestational age. Double immunohistochemistry using antibodies to CD14 and CD68 was performed, and the density of double or single positive cells in the basal plate and placental bed was evaluated. Non-parametric statistics were used for analysis. Results 1) A unique subset of CD14−/CD68+ cells was identified. The cells in question were present at a higher level in the decidua than in the myometrial segment of the placental bed ( p < 0.01); 2) The density and proportion of CD14+/CD68+ cells (double positive cells) were significantly higher in the myometrial segment than in the basal plate ( p = 0.0003); and 3) There were no significant differences in the density and patterns of immunopositive macrophages in the basal plate, the decidua, and the myometrium between women with preeclampsia and those with preterm labor/delivery ( p > 0.05). Conclusion The macrophages at the fetomaternal interface can be dichotomized by CD14 and CD68 immunoreactivity. A gradient of CD14+/CD68+ macrophages was demonstrated between the superficial myometrium and the basal plate regardless of the etiology of preterm birth (preeclampsia or spontaneous preterm labor). The biological function of single positive (CD14−/CD68+) and double positive (CD14+/CD68+) macrophages at the fetomaternal interface remains to be established. The overall findings also suggest that the discrepancies in the literature are due to the varying markers used to detect macrophages and in the anatomical plane of the fetomaternal junction analyzed.

Effect of Vitamin E and selenium supplementation on concentrations of plasma cortisol and erythrocyte lipid peroxides and the incidence of retained fetal membranes in crossbred dairy cattle

The objectives were to: (i) determine the effect of prepartum supplementation of Vitamin E (Vit E) and selenium (Se) on plasma cortisol, erythrocyte peroxidation and the incidence of retained fetal membranes (RFM); (ii) estimate myeloperoxidase (MPO), lysozyme, elastase, and acid phosphatase (ACP) enzyme activities in the cotyledons of cows with or without RFM; and (iii) determine the molecular weight (SDS-PAGE) of proteins present in the cotyledons of cows with or without RFM. Fifty dairy (Friesian × Sahiwal) cows were equally allocated to one of two treatments, given as an im injection 3 week before calving: 1100 IU of DL α-tocopherol acetate (Vit E) and 30 mg of sodium selenite (Se), or saline (control). Concentrations of plasma cortisol (20 cows) were determined on days 21, 7, 3, 2, 1, and 0 prepartum, and erythrocyte lipid peroxide (all cows) was determined on days 21 and 7 prepartum. Treatment with Vit E and Se did not affect ( P = 0.23) the incidence of RFM (12% versus 0%, respectively) but decreased ( P < 0.05) erythrocyte lipid peroxide concentrations on day 7 prepartum compared with day 21 prepartum. Plasma cortisol concentration increased ( P < 0.05) from day 21 prepartum to the day of parturition in Vit E + Se and control cows. However, on day 0, plasma cortisol concentrations were lower ( P < 0.05) in cows given Vit E + Se than in control cows (with or without RFM). To investigate enzyme activity and peptides in cotyledons, cotyledons were collected (from cows that were not part of the principal experiment), homogenised with PBS, and the supernatant used for the estimation of cationic peptides. Cotyledons of cows with RFM ( n = 8) had lower ( P < 0.01) MPO and greater ( P < 0.05) lysozyme and ACP enzyme activities than those from non-RFM cows ( n = 6). A band at < 10 kDa in the SDS-PAGE indicated the presence of cationic peptides. In conclusion, a single treatment of Vit E and Se at 3-week prepartum reduced concentrations of plasma cortisol and erythrocyte peroxide. Altered enzyme activities in the fetal membranes indicated the involvement of leukocytes and trauma at the fetomaternal junction and warrant further investigation.

The pathogenesis of pre-eclampsia: new aspects.

The etiology of pre-eclampsia, a disorder specific to pregnancy, has not yet been clearly established. Generalized endothelial dysfunction is thought to occur. Inadequate trophoblast invasion at the feto-maternal junction has also been postulated as the cause. However, recent findings are more suggestive of an inappropriate maternal inflammatory response within the framework of placentation, the innate immune system being primarily involved. This exaggerated maternal intravascular inflammatory reaction to the invading trophoblast leads to the manifestations of pre-eclampsia in the mother and feto-placental unit. The extent of the inflammatory reaction, which is also present in normal pregnancy, is probably determined by genetic factors and any pre-existing disorder affecting the mother or fetus. The process is very complex and cannot be attributed to any one single cause.

Immunohistological and biochemical evidence for a role for hyaluronic acid in the growth and development of the placenta

A monoclonal antibody, designated NDOG1, has been used to stain a series of human and monkey placentae as well as several adult human tissues using immunoperoxidase techniques. In early placentae, NDOG1 was found to stain extracellular material associated with proliferating, extravillous cytotrophoblast cell columns and with the cytotrophoblast shell at the feto-maternal junction. The immunohistology suggests that NDOG1 antigen may be secreted by the anchoring cytotrophoblast into the immediately adjacent maternal tissues. NDOG1 antibody also shows extracellular staining in the stroma of early human placentae and reacted with the apical villous syncytiotrophoblast plasma membrane throughout pregnancy. Biochemical experiments demonstrated that extracts of this latter membrane contained NDOG1 antigenic activity which was susceptible to digestion with bovine testicular hyaluronidase. Hyaluronic acid was the only glycosaminoglycan found in this membrane, thereby implying a reaction between NDOG1 antibody and hyaluronic acid. Whilst no such direct interaction could be demonstrated in vitro. NDOG1 was shown to compete with two other antibodies which themselves demonstrated specificity for hyaluronic acid. The proposed identity between the NDOG1 antigen and hyaluronic acid is discussed particularly in terms of placentation where the distribution of NDOG1 staining may confirm the role of hyaluronic acid in providing an open matrix structure during stages of cell proliferation, migration and invasion.

Ultrastructural observations on the chorioallantoic placenta of the golden‐rumped elephant shrew, Rhynchocyon chrysopygus

SummaryThe chorioallantoic placenta of the elephant shrew (Rhynchocyon) was studied at the ultrastructural level. The placenta is haemonochorial with an appparent lack of microvilli on the syncytiotrophoblastic surface. There is, however, an unusual development of subsurface bays which are covered only by an ectoplasmic layer of trophoblast. The contents of the bays is PAS‐negative and is similar in both texture and appearance to the maternal blood plasma. The basal trophoblastic cells are exceptionally tall and are held together by numerous desmosomes. Their basal lamina, which is on the trophospongial side, it unusually thick. At the fetomaternal junction the basal trophoblastic cells are coated with a finely granular electron‐dense precipitate of unknown nature.Electron‐dense particles resembling iron deposits are demonstrable in the fetal mesenchyme of the labyrinthine zone.

Electron microscopic study of the chorioallantoic placenta of the rock hyrax (Heterohyrax brucei).

The interhaemal membrane consisted of only two cellular elements: a single layer of cellular trophoblast and the fetal capillary endothelium. The hyrax is therefore one of the few mammals known to possess the cellular haemomonochorial type of placenta. The trophospongium was also cellular while the basal trophoblastic cells were strongly phagocytic. The giant multinucleate cells at the feto-maternal junction were ultrastructurally different from the trophoblast cells and showed no signs of degeneration. Their appearance suggests that they are of maternal rather than fetal origin.

Autoradiographic evidence for migration and fusion of cells in the sheep placenta: Resolution of a problem in placental classification

The location of the fetomaternal junction in the placenta is important with respect to structural classification and the identification of any possible barrier to maternal immunorejection. Structural classification of the sheep placenta remains controversial on account of the uncertain origin of the syncytial layer. In this study [ 3H]thymidine was injected into the fetus and placentomes were removed between 4h and 21 days afterwards. Autoradiography showed that the syncytium is derived predominantly from the migration of fetal binucleate cells and not from the maternal uterine epithelium as most recent reports have suggested. In this respect the origin of the syncytium in the sheep placenta is similar to that reported in certain other eutherian mammals. The finding that cells originating from the fetal allograft survive after migration through the microvillous junction poses questions as to the mechanism by which the syncytial layer resists maternal immune rejection throughout gestation.

Immuno and histochemical electron microscopic studies of the selective permeability, involving the feto-maternal junctional barrier

Regarding to the cellular transfer and absorption mechanism of the trophoblast in the feto-maternal junctional zone, it is awaited futher confirmation in the future. Numerous documentation has been accumulated on this aspect, utilizing the visible markers such as the fat(Yamaguchi), ferritin(Tillack) and IgG(Morphis). however, the observations are mainly based on the investigations of the placenta in vitro. From this view point, the authors have made a rather comprehensive observation of the cellular transfer and absorption mechanism of the human feto-maternal junction in physiological circumstance.Materials and methods: The specimens of the human uterus in early ges tation are removed surgically for therapeutic purpose had served as materials. The chorionic tissue and implanted tissue from the uterus were carefully investigated. The method of the investigation are as follows: 1) Conventional transmission electron microscopic observation with double electron stains. 2) Visualization of the iron dextran(Fesin), it is usually used to iron deficiency anemia, which was administered prior to the surgical removal of the uterus.

The role of placental gonadotrophins (PMSG and hCG) in pregnancy in the rat.

Treatment of rats with anti-PMSG and anti-hCG for 5 days from Days 6, 11 or 16 of pregnancy caused variable necrosis of fetal tissue, the extent of which increased from the chorionic cells to the inner trophoblastic layers with the increase in dose from 4 to 10 mg anti-PMSG. At 10 mg/day, fetal death and resorption were invariable in early to mid-pregnancy, whilst in late pregnancy some litters were carried to term. The main histological change in the corpora lutea was hyperaemia in the anti-PMSG-treated rats. Immunofluorescence studies with conjugates of anti-hCG and anti-PMSG showed sharp localization in the giant and chorionic cell layer at the feto-maternal junction. We conclude that an hCG/PMSG-like substance is produced by rat chorionic cells and that it plays an immunosuppressive role during pregnancy.