scholarly journals Spatiotemporal Expression of C-CAM in the Rat Placenta

1997 ◽  
Vol 45 (7) ◽  
pp. 1021-1034 ◽  
Author(s):  
Hiroki Sawa ◽  
Hiroyuki Ukita ◽  
Minoru Fukuda ◽  
Hajime Kamada ◽  
Isamu Saito ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Broad Band ◽  
Placental Tissue ◽  
Maternal Blood ◽  
Immunoglobulin Superfamily ◽  
Pcr Analysis ◽  
Spatiotemporal Pattern ◽  
Positive Staining ◽  
Placental Development ◽  
Embryonic Vessels

We investigated the expression of the immunoglobulin superfamily cell adhesion molecule, C-CAM, in developing and mature rat placenta. By immunohistochemical staining at the light microscopic level, no C-CAM-expression was seen before Day 9 of gestation, when it appeared in the trophoblasts of ectoplacental cones. On Day 10.5, spongiotrophoblasts and invasive trophoblasts around the maternal vessels of the decidua basalis were stained positively. On Day 12.5, C-CAM was detected in the spongiotrophoblasts of the junctional layer, but labyrinth trophoblasts and secondary giant trophoblasts were not stained. On Day 17.5, C-CAM was found only in the labyrinth and lacunae of the junctional layer. At this stage, both the labyrinth cytotrophoblasts of the maternal blood vessels and the endothelial cells of the embryonic capillaries were strongly stained. Placental tissues from gestational Days 12.5 and 17.5 were analyzed by immunoelectron microscopy to determine the location of C-CAM at the subcellular level. On Day 12.5, positive staining of the spongiotrophoblasts was observed, mainly on surface membranes and microvilli between loosely associated cells. On Day 17.5, staining was found primarily on the microvilli of the maternal luminal surfaces of the labyrinth cytotrophoblasts, and both on the luminal surface and in the cytoplasm of endothelial cells of the embryonic vessels. RT-PCR analysis and Southern blotting of the PCR products revealed expression of mRNA species for both of the major isoforms, C-CAM1 and C-CAM2. Immunoblotting analysis of C-CAM isolated from 12.5-day and 14.5-day placentae showed that it appeared as a broad band with an apparent molecular mass of 110–170 kD. In summary, C-CAM was strongly expressed in a specific spatiotemporal pattern in trophoblasts actively involved in formation of the placental tissue, suggesting an important role in placental development. In the mature placenta, C-CAM expression was confined to the trophoblastic and endothelial cells lining the maternal and embryonic vessels, respectively, suggesting important functions in placental physiology.

Florid Bacillus cereus Infection of the Placenta Associated With Intrauterine Fetal Demise

2021 ◽  
pp. 109352662199902
Author(s):  
Stephanie Shea ◽  
Alberto Paniz-Mondolfi ◽  
Emilia Sordillo ◽  
Michael Nowak ◽  
Fumiko Dekio
Keyword(s):  
Bacillus Cereus ◽  
Placental Tissue ◽  
Maternal Blood ◽  
Gastrointestinal Infections ◽  
Gram Positive ◽  
Fetal Demise ◽  
Placental Infection ◽  
Intrauterine Fetal Demise ◽  
Acute Necrotizing ◽  
Poor Outcomes

Bacillus cereus is a gram-positive, rod-shaped bacterium that is commonly implicated in foodborne illness but has also become increasingly recognized as a source of serious non-gastrointestinal infections, including sepsis, meningitis, and pneumonia. Non-gastrointestinal B. cereus infections have been identified in children, especially in neonates; however, there are no previously described cases of fetal demise associated with B. cereus placental infection. We present a case of acute chorioamnionitis-related intrauterine fetal demise of twin A at 17 weeks gestation, noted two days after selective termination of twin B. Histological examination revealed numerous gram-positive bacilli in placental tissue, as well as fetal vasculature, in the setting of severe acute necrotizing chorioamnionitis and subchorionitis, intervillous abscesses, acute villitis, and peripheral acute funisitis. Cultures of maternal blood and placental tissue both yielded growth of B. cereus. This case underscores the importance of B. cereus as a human pathogen, and specifically demonstrates its potential as an agent of severe intraamniotic and placental infection with poor outcomes for the fetus.

Abstract 294: Expression of Matrix Metalloproteases during the Differentiation of Porcine Adopse-Drived Mesenchymal Stem Cells ro Endothelial Cells

2013 ◽  
Vol 33 (suppl_1) ◽  
Author(s):  
Sami G Almalki ◽  
Velidi Rao ◽  
Divya Pankajakshan ◽  
Devendra K Agrawal
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Mesenchymal Stem Cells ◽  
Enzyme Activity ◽  
Protein Expression ◽  
Western Blot ◽  
Gelatin Zymography ◽  
Matrix Metalloproteases ◽  
Positive Staining ◽  
Mrna Transcripts

Rationale Adipose-derived mesenchymal stem cells (ADMSCs) are multipotent cells that have the potential to differentiate into different cell linages, and represent promising tools in various clinical applications. However, the molecular mechanisms that control the ability of ADMSCs to remodel 3-dimensional extracellular matrix (ECM) barriers during differentiation are not clearly understood. Herein, we studied the expression of matrix metalloproteinases (MMPs) during the differentiation of ADMSCs to endothelial cells (ECs) in vitro . Methods MSCs were isolated from porcine abdominal adipose tissue, and characterized by positive staining for MSC markers, CD44, CD73, CD90, and negative staining for CD11b, CD34 and CD45. The plasticity of MSCs was detected by bi-lineage differentiation to osteocytes, and adipocytes. The mRNA transcripts for different MMPs and TIMPs and protein expression of EC markers were analyzed by RT-PCR and immunostaining. The enzyme activity and protein expression were also analyzed by gelatin zymography, ELISA, and Western blot. Results The differentiation of ADMSCs to ECs was confirmed by the positive staining and mRNA expression of the endothelial markers. The mRNA transcripts for MMP-2 and membrane type 1 MMP (MT1-MMP) was significantly increased by 2.5 and 2.0 fold, respectively, during the differentiation of MSCs into ECs. Western blot and ELISA showed an elevated MT1-MMP and MMP-2 expression. The enzyme activity of MMP-2 was also observed by gelatin zymography. Conclusion We demonstrated that porcine ADMSCs have the ability to differentiate into ECs, and this process involves the up-regulation of MMP-2 and MT1-MMP. The increase in the expression of MMP-2 and MT1-MMP may, at least partially, facilitate the change in morphology of MSCs by degrading the ECM barriers. These findings may provide a potential mechanism for the role of MMP2 and MT1-MMP in the differentiation of ADMSCs into ECs.

scholarly journals Spatiotemporal Pattern of Neuroinflammation After Impact-Acceleration Closed Head Injury in the Rat

2006 ◽  
Vol 2006 ◽  
pp. 1-6 ◽  
Author(s):  
Servan Rooker ◽  
Sebastian Jander ◽  
Jos Van Reempts ◽  
Guido Stoll ◽  
Philippe G. Jorens ◽  
...  
Keyword(s):  
Head Injury ◽  
Interleukin 1 ◽  
Closed Head Injury ◽  
Brain Structures ◽  
Pcr Analysis ◽  
Spatiotemporal Pattern ◽  
Inos Mrna ◽  
Rapid Induction ◽  
Closed Head ◽  
Primary Impact

Inflammatory processes have been implicated in the pathogenesis of traumatic brain damage. We analyzed the spatiotemporal expression pattern of the proinflammatory key molecules: interleukin-1β, interleukin-6, tumor necrosis factor-α, and inducible nitric oxide synthase in a rat closed head injury (CHI) paradigm. 51 rats were used for RT-PCR analysis after CHI, and 18 for immunocytochemistry. We found an early upregulation of IL-1β, IL-6, and TNF-αmRNA between 1 h and 7 h after injury; the expression of iNOS mRNA only revealed a significant increase at 4 h. After 24 h, the expression decreased towards baseline levels, and remained low until 7 d after injury. Immunocytochemically, IL-1βinduction was localized to ramified microglia in areas surrounding the primary impact place as well as deeper brain structures. Our study shows rapid induction of inflammatory gene expression that exceeds by far the primary impact site and might therefore contribute to tissue damage at remote sites.

scholarly journals Amelioration of C - Reactive Protein and Lectin Like Oxidised Low Density Lipoprotein Receptor Complex Induced Endothelial Dysfunction by Oligomeric Proanthocyanidins.

2021 ◽  
Author(s):  
Sankar Jamuna ◽  
Rathinavel Ashokkumar ◽  
Niranjali Devaraj Sivasithamparam
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Endothelial Dysfunction ◽  
Capillary Tube ◽  
Morphological Changes ◽  
Protein Docking ◽  
Pcr Analysis ◽  
Inhibitory Mechanism ◽  
C Reactive Protein ◽  
Reactive Protein

Abstract C-reactive protein (CRP) is a well established biochemical marker for atherosclerosis. Inflammation induced by CRP promotes endothelial dysfunction. Modification of LDL inside the artery wall favors the elevation of this acute phase protein. The mechanism of OxLDL+CRP complex is unrevealed so far. Hence, this mechanism was considered as the important factor to trigger the monocyte to macrophages differentiation which in leads to foam cells formation. Hence this key event should be targeted and focused on how this complex (OxLDL+CRP) proceeds to endothelial dysfunction. OPC is a well known cardioprotective flavon-3-ols. The present study is challenged between the protective roles of OPC against the deleterious effect of this complex (OxLDL+CRP) on endothelial cells. Monolayer of Endothelial cells were incubated with THP-1 monocytes for 48 h supplemented with OxLDL (10mg/ml) + CRP (10 mg/ml) complex and treated with OPC (100mg/ml). Morphological changes, cell migration assay and capillary tube forming assay was carried out. Myeleoperoxidase levels were estimated to determine the adhesion of monocytes onto EC monolayer. RT-PCR analysis of L-Selectin was done. The quantification of NO levels and analysis of mRNA expressions of eNOS is to determine the nitric oxide demand caused due to OxLDL+CRP complex. LOX-1, scavenger receptor levels were analysed by mRNA expression. Proinflammatory markers such as IL-6, MCP-1 and IL-1b were studied. Accumulation of ROS levels were measured fluorimetrically using DCF-DA. Spectrophotometric analysis of Sirius red dye binding collagen levels was observed. Mitochondrial membrane potential was determined by JC-1 dye and cell cycle analysis was done by FACS analysis. Protein –Protein docking was carried out between CRP and LOX-1. This docked protein complex were again docked with OPC and atrovastatin to show the inhibitory mechanism of CRP binding with LOX-1. OPC showed a promising inhibitory mechanism against OxLDL+CRP complex. To emphasis the results OPC treated group showed decreased levels of proinflammatory markers, LOX-1 and L-Selectin levels. Endothelial nitric oxide levels were increased upon OPC treatment and reduction in the ROS levels. Endothelial cells apoptosis was prevented by OPC. Docking studies showed that in the absence of ligands (OPC) binding of CRP and LOX-1 was greater and vice versa in the presence of ligands. To conclude, OxLDL + CRP complex inhibitory effects of OPC could maintain the normal homeostasis.

scholarly journals CD63 is a component of Weibel-Palade bodies of human endothelial cells

Blood ◽  
1993 ◽  
Vol 82 (4) ◽  
pp. 1184-1191 ◽  
Author(s):  
UM Vischer ◽  
DD Wagner
Keyword(s):  
Endothelial Cells ◽  
Cell Surface ◽  
Western Blotting ◽  
Vascular Endothelial Cells ◽  
Secretory Granules ◽  
Positive Staining ◽  
Cell Fractionation ◽  
Glycosylation Pattern ◽  
Adhesion Proteins ◽  
Shape Distribution

Weibel-Palade bodies are secretory granules of vascular endothelial cells specialized in the storage of von Willebrand factor (vWF) and P- selectin, two adhesion proteins that can be rapidly mobilized to the cell surface by exocytosis in response to thrombin or other agonists. In this study, we attempted to identify additional components of Weibel- Palade bodies by raising monoclonal antibodies to these granules, purified by cell fractionation. One antibody, 2C6, was found to be specific for CD63, a membrane glycoprotein previously described in the lysosomes of platelets and other cell types. The immunopurified 2C6 antigen was recognized by an anti-CD63 reference antibody, 2.28, by Western blotting. Also, the biosynthetic profile of the 2C6 antigen in endothelial cells showed a nascent molecular mass and a glycosylation pattern identical to that of CD63. Immunofluorescence staining with 2C6 showed the lysosomes, and also elongated structures identified as Weibel-Palade bodies by their shape, distribution, and positive staining with anti-vWF antibodies, CD63 was also found by Western blotting of subcellular fractions highly enriched in Weibel-Palade bodies. Our results indicate that CD63 colocalizes with vWF and P- selectin in the Weibel-Palade bodies of endothelial cells, and together with these adhesion proteins it could be rapidly expressed on the cell surface in areas of vascular injury and inflammation.

scholarly journals Study of Human T21 Placenta Suggests a Potential Role of Mesenchymal Spondin-2 in Placental Vascular Development

Endocrinology ◽  
2019 ◽  
Vol 160 (3) ◽  
pp. 684-698 ◽  
Author(s):  
Pascale Gerbaud ◽  
Padma Murthi ◽  
Jean Guibourdenche ◽  
Fabien Guimiot ◽  
Benoît Sarazin ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Biochemical Analysis ◽  
Mesenchymal Cells ◽  
Conditioned Media ◽  
Chromosome 21 ◽  
Placental Development ◽  
Cm Protein ◽  
Protein Array Analysis ◽  
Functional Analyses

Abstract Placental development is particularly altered in trisomy of chromosome 21 (T21)–affected pregnancies. We previously described in T21-affected placentae an abnormal paracrine crosstalk between the villus mesenchymal core and villus trophoblasts. T21-affected placentae are known to be characterized by their hypovascularity. However, the causes of this anomaly remain not fully elucidated. Therefore, the hypothesis of an abnormal paracrine crosstalk between fetal mesenchymal core and placental endothelial cells (PLECs) was evocated. Villus mesenchymal cells from control (CMCs) and T21 placentae (T21MCs) were isolated and grown in culture to allow their characterization and collection of conditioned media for functional analyses (CMC-CM and T21MC-CM, respectively). Interestingly, PLEC proliferation and branching ability were less stimulated by T21MC-CM than by CMC-CM. Protein array analysis identified secreted proangiogenic growth factors in CMC-CM, which were reduced in T21MC-CM. Combined mass spectrometry and biochemical analysis identified spondin-2 as a factor decreased in T21MC-CM compared with CMC-CM. We found that exogenous spondin-2 stimulated PLEC proliferation and established that T21MC-CM supplemented with spondin-2 recovered conditioned media ability to induce PLEC proliferation and angiogenesis. Hence, this study demonstrates a crosstalk between villus mesenchymal and fetal endothelial cells, in which spondin-2 secreted from mesenchymal cells plays a central role in placental vascular functions. Furthermore, our results also suggest that a reduction in spondin-2 secretion may contribute to the pathogenesis of T21 placental hypovascularity.

ULTRASTRUCTURAL AND Y-PCR ANALYSIS OF TROPHOBLAST-LIKE CELLS DETECTED IN PERIPHERAL MATERNAL BLOOD USING ANTIBODIES AND FLOW CYTOMETRY SORTING

1991 ◽  
Vol 73 (2-3) ◽  
pp. 34a-34a
Author(s):  
Philippe Metezeau ◽  
Jean-Frédéric Bruch ◽  
Nicole Garcia-Fonknechten ◽  
Hélène Kiefer ◽  
Christophe Julien ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Maternal Blood ◽  
Pcr Analysis ◽  
Flow Cytometry Sorting

Role of endogenous digitalis-like factors in the clinical manifestations of severe preeclampsia: a systematic review

2018 ◽  
Vol 132 (12) ◽  
pp. 1215-1242 ◽  
Author(s):  
Vardaman M. Buckalew
Keyword(s):  
Antihypertensive Drugs ◽  
Post Partum ◽  
Secondary Analysis ◽  
Clinical Manifestations ◽  
Antibody Fragment ◽  
Placental Tissue ◽  
Maternal Blood ◽  
Double Blind ◽  
Red Cell Membrane ◽  
Natriuretic Hormone

Endogenous digitalis-like factor(s), originally proposed as a vasoconstrictor natriuretic hormone, was discovered in fetal and neonatal blood accidentally because it cross-reacts with antidigoxin antibodies (ADAs). Early studies using immunoassays with ADA identified the digoxin-like immuno-reactive factor(s) (EDLF) in maternal blood as well, and suggested it originated in the feto–placental unit. Mammalian digoxin-like factors have recently been identified as at least two classes of steroid compounds, plant derived ouabain (O), and several toad derived bufodienolides, most prominent being marinobufagenin (MBG). A synthetic pathway for MBG has been identified in mammalian placental tissue. Elevated maternal and fetal EDLF, O and MBG have been demonstrated in preeclampsia (PE), and inhibition of red cell membrane sodium, potassium ATPase (Na, K ATPase (NKA)) by EDLF is reversed by ADA fragments (ADA-FAB). Accordingly, maternal administration of a commercial ADA-antibody fragment (FAB) was tested in several anecdotal cases of PE, and two, small randomized, prospective, double-blind clinical trials. In the first randomized trial, ADA-FAB was administered post-partum, in the second antepartum. In the post-partum trial, ADA-FAB reduced use of antihypertensive drugs. In the second trial, there was no effect of ADA-FAB on blood pressure, but the fall in maternal creatinine clearance (CrCl) was prevented. In a secondary analysis using the pre-treatment maternal level of circulating Na, K ATPase (NKA) inhibitory activity (NKAI), ADA-FAB reduced the incidence of pulmonary edema and, unexpectedly, that of severe neonatal intraventricular hemorrhage (IVH). The fall in CrCl in patients given placebo was proportional to the circulating level of NKAI. The implications of these findings on the pathophysiology of the clinical manifestations PE are discussed, and a new model of the respective roles of placenta derived anti-angiogenic (AAG) factors (AAGFs) and EDLF is proposed.

203. A differential pattern of follistatin expression in the placenta between spontaneous, induced and non-labouring patient groups

2005 ◽  
Vol 17 (9) ◽  
pp. 77
Author(s):  
K. M. Rae ◽  
K. G. Hollebone ◽  
L. Meng ◽  
D. C. Clausen ◽  
J. R. McFarlane
Keyword(s):  
Vascular Endothelial Cells ◽  
Placental Tissue ◽  
Cell Types ◽  
Cross Sectional Study ◽  
Polyclonal Antiserum ◽  
Antigen Retrieval ◽  
Positive Staining ◽  
Cross Sectional ◽  
Chorionic Villi ◽  
Patient Groups

Follistatin has been identified in human placenta, fetal membranes and fluids, with serum follistatin concentrations rising during pregnancy, particularly near term. Our laboratory has shown follistatin concentrations rise across labour in spontaneous but not induced women.1 As the placenta is a source of follistatin, this study examined placental tissues using immunohistochemistry to determine differences in follistatin localization between groups. Placental tissue was collected immediately following delivery from three groups of women at term, spontaneous onset (n = 4), induction (n = 4) and non-labouring caesarian (n = 4), and immediately formalin fixed. Antigen-retrieval immunohistochemistry using a specific chicken polyclonal antiserum (CK20) raised against a follistatin peptide (AA 121-133) or pre-immune chicken serum was performed. Positive staining of syncytiotrophoblast cells of the chorionic villi was seen in patients undergoing spontaneous labour but not in the induced and caesarian delivery group. The two labouring groups (spontaneous and induced) both showed positive staining for the vascular endothelial cells within the chorionic villi and the stratum basale, whilst the caesarian delivery group was negative for any staining within these vessels. Positive staining of Hofbauer cells was observed in both labouring groups; however, the caesarian group showed infrequent positive staining of these cell types. The differences in expression pattern in the two labouring groups (spontaneous v. induced) may be due to variations in labour lengths (6.5 v. 4.5 h, respectively); however, we would have expected a lower level of expression in the same cell types rather than the complete absence of staining. The positive follistatin staining in the syncytiotrophoblast of spontaneous patients suggests this may be the source of the rising plasma follistatin seen in this group. These differences in staining support our hypothesis that an earlier endocrine signal is absent in the induced and caesarian patient groups. (1)Rae K, Hollebone K, Clausen DC, Chetty V, McFarlane JR. (2004). A Cross-Sectional Study of Follistatin During Labour in Women. The Endocrine Societies 86th Annual Meeting, New Orleans, 2004.

scholarly journals Transcription Factor PLAGL1 Is Associated with Angiogenic Gene Expression in the Placenta

2020 ◽  
Vol 21 (21) ◽  
pp. 8317
Author(s):  
Rebekah R. Starks ◽  
Rabab Abu Alhasan ◽  
Haninder Kaur ◽  
Kathleen A. Pennington ◽  
Laura C. Schulz ◽  
...  
Keyword(s):  
Gene Expression ◽  
Endothelial Cells ◽  
Transcription Factor ◽  
Gene Networks ◽  
Critical Role ◽  
Placental Development ◽  
Human Trophoblast ◽  
Mouse Placenta ◽  
Expression Of Genes ◽  
Sirna Knockdown

During pregnancy, the placenta is important for transporting nutrients and waste between the maternal and fetal blood supply, secreting hormones, and serving as a protective barrier. To better understand placental development, we must understand how placental gene expression is regulated. We used RNA-seq data and ChIP-seq data for the enhancer associated mark, H3k27ac, to study gene regulation in the mouse placenta at embryonic day (e) 9.5, when the placenta is developing a complex network of blood vessels. We identified several upregulated transcription factors with enriched binding sites in e9.5-specific enhancers. The most enriched transcription factor, PLAGL1 had a predicted motif in 233 regions that were significantly associated with vasculature development and response to insulin stimulus genes. We then performed several experiments using mouse placenta and a human trophoblast cell line to understand the role of PLAGL1 in placental development. In the mouse placenta, Plagl1 is expressed in endothelial cells of the labyrinth layer and is differentially expressed in placentas from mice with gestational diabetes compared to placentas from control mice in a sex-specific manner. In human trophoblast cells, siRNA knockdown significantly decreased expression of genes associated with placental vasculature development terms. In a tube assay, decreased PLAGL1 expression led to reduced cord formation. These results suggest that Plagl1 regulates overlapping gene networks in placental trophoblast and endothelial cells, and may play a critical role in placental development in normal and complicated pregnancies.

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