Placental vascularisation requires the AP-1 component fra1

Development ◽  
2000 ◽  
Vol 127 (22) ◽  
pp. 4937-4948 ◽  
Author(s):  
M. Schreiber ◽  
Z.Q. Wang ◽  
W. Jochum ◽  
I. Fetka ◽  
C. Elliott ◽  
...  
Keyword(s):  
Marked Decrease ◽  
Vascular Endothelial Cells ◽  
Es Cells ◽  
Early Gene ◽  
Marker Genes ◽  
Vascular Endothelial ◽  
Wild Type ◽  
Mouse Development ◽  
Gene Encoding ◽  
Embryonic Tissues

Fra1 is an immediate-early gene encoding a member of the AP-1 transcription factor family, which has diverse roles in development and oncogenesis. To determine the function of Fra1 in mouse development, the gene was inactivated by gene targeting. Foetuses lacking Fra1 were severely growth retarded and died between E10.0 and E10.5, owing to defects in extra-embryonic tissues. The placental labyrinth layer, where X-gal staining revealed expression of Fra1, was reduced in size and largely avascular, owing to a marked decrease in the number of vascular endothelial cells, as shown by the lack of Flk1 expression. In contrast, the spongiotrophoblast layer was unaffected and expressed the marker genes 4311 (Tpbp) and Flt1. Furthermore, mutant foetuses exhibited yolk-sac defects that may contribute to their growth retardation and lethality. Importantly, when the placental defect was rescued by injection of Fra1(−)(/)(−) ES cells into tetraploid wild-type blastocysts, Fra1(−)(/)(−) pups were obtained that were no longer growth retarded and survived up to 2 days after birth without apparent phenotypic defects. These data indicate that a defect in the extra-embryonic compartment is causal to the observed lethality, and suggest that Fra1 plays a crucial role in establishing normal vascularisation of the placenta.

scholarly journals Activation of TF-Dependent Blood Coagulation Pathway and VEGF-A in Patients with Essential Thrombocythemia

Medicina ◽  
2019 ◽  
Vol 55 (2) ◽  
pp. 54
Author(s):  
Grażyna Gadomska ◽  
Katarzyna Ziołkowska ◽  
Joanna Boinska ◽  
Jan Filipiak ◽  
Danuta Rość
Keyword(s):  
Tissue Factor ◽  
Essential Thrombocythemia ◽  
Vascular Endothelial Cells ◽  
Myeloproliferative Neoplasms ◽  
Simultaneous Increase ◽  
Control Group ◽  
Vascular Endothelial ◽  
Gene Encoding ◽  
Positive Correlation ◽  
Normal Individuals

Background and objectives: Recent studies suggest that a vascular endothelial growth factor (VEGF-A) may be involved in the thrombotic process by stimulating the expression of tissue factor in vascular endothelial cells. Tissue factor (TF) can also stimulate the transcription of the gene encoding VEGF-A. The relationship between coagulation and angiogenesis in myeloproliferative neoplasms is not fully understood. The aim of this study was to evaluate the concentration of TF in relation to VEGF-A in the blood of patients with essential thrombocythemia (ET). Patients and methods: The study group consisted of 130, newly diagnosed patients with ET (mean age 61 years). The control group consisted of 35 healthy volunteers (mean age 51 years). Concentrations of VEGF-A, TF, and tissue factor pathway inhibitor (TFPI) were analysed using immunoenzymatic methods. TF and TFPI activities were performed using chromogenic assays. Results: The median concentration of TF Ag was 3-fold higher and the TF activity was more than 15-fold higher in ET patients than in normal individuals. There were no statistically significant differences in the TFPI concentration and activity between groups. VEGF-A was significantly increased in patients with ET (p < 0.000001). Analysis of correlations revealed a positive correlation between VEGF-A and TF Ag as well as a positive correlation between VEGF-A and TFPI activity. Conclusions: The simultaneous increase of TF concentration and activity, VEGF-A in the blood of patients with ET, as well as a positive correlation between the concentration of TF and VEGF-A demonstrates the coexistence of TF-dependent coagulation and activation of angiogenesis.

scholarly journals The Effect of Sodium Bicarbonate, a Beneficial Adjuvant Molecule in Cystic Fibrosis, on Bronchial Epithelial Cells Expressing a Wild-Type or Mutant CFTR Channel

2020 ◽  
Vol 21 (11) ◽  
pp. 4024 ◽  
Author(s):  
Ilona Gróf ◽  
Alexandra Bocsik ◽  
András Harazin ◽  
Ana Raquel Santa-Maria ◽  
Gaszton Vizsnyiczai ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Endothelial Cells ◽  
Epithelial Cells ◽  
Sodium Bicarbonate ◽  
Vascular Endothelial Cells ◽  
Bronchial Epithelium ◽  
Barrier Properties ◽  
Vascular Endothelial ◽  
Wild Type ◽  
Bronchial Epithelial

Clinical and experimental results with inhaled sodium bicarbonate as an adjuvant therapy in cystic fibrosis (CF) are promising due to its mucolytic and bacteriostatic properties, but its direct effect has not been studied on respiratory epithelial cells. Our aim was to establish and characterize co-culture models of human CF bronchial epithelial (CFBE) cell lines expressing a wild-type (WT) or mutant (deltaF508) CF transmembrane conductance regulator (CFTR) channel with human vascular endothelial cells and investigate the effects of bicarbonate. Vascular endothelial cells induced better barrier properties in CFBE cells as reflected by the higher resistance and lower permeability values. Activation of CFTR by cAMP decreased the electrical resistance in WT but not in mutant CFBE cell layers confirming the presence and absence of functional channels, respectively. Sodium bicarbonate (100 mM) was well-tolerated by CFBE cells: it slightly reduced the impedance of WT but not that of the mutant CFBE cells. Sodium bicarbonate significantly decreased the more-alkaline intracellular pH of the mutant CFBE cells, while the barrier properties of the models were only minimally changed. These observations indicate that sodium bicarbonate is beneficial to deltaF508-CFTR expressing CFBE cells. Thus, sodium bicarbonate may have a direct therapeutic effect on the bronchial epithelium.

scholarly journals Deficiency of the Stress Kinase P38α Results in Embryonic Lethality

2000 ◽  
Vol 191 (5) ◽  
pp. 859-870 ◽  
Author(s):  
Melanie Allen ◽  
Linne Svensson ◽  
Marsha Roach ◽  
John Hambor ◽  
John McNeish ◽  
...  
Keyword(s):  
Map Kinase ◽  
Es Cells ◽  
Interleukin 1 ◽  
Embryonic Stem ◽  
Wild Type ◽  
Antiinflammatory Drugs ◽  
Gene Encoding ◽  
Significant Difference ◽  
Induced Apoptosis

The mitogen-activated protein (MAP) kinase p38 is a key component of stress response pathways and the target of cytokine-suppressing antiinflammatory drugs (CSAIDs). A genetic approach was employed to inactivate the gene encoding one p38 isoform, p38α. Mice null for the p38α allele die during embryonic development. p38α1/− embryonic stem (ES) cells grown in the presence of high neomycin concentrations demonstrated conversion of the wild-type allele to a targeted allele. p38α−/− ES cells lacked p38α protein and failed to activate MAP kinase–activated protein (MAPKAP) kinase 2 in response to chemical stress inducers. In contrast, p38α1/+ ES cells and primary embryonic fibroblasts responded to stress stimuli and phosphorylated p38α, and activated MAPKAP kinase 2. After in vitro differentiation, both wild-type and p38α−/− ES cells yielded cells that expressed the interleukin 1 receptor (IL-1R). p38α1/+ but not p38α−/− IL-1R–positive cells responded to IL-1 activation to produce IL-6. Comparison of chemical-induced apoptosis processes revealed no significant difference between the p38α1/+ and p38α−/− ES cells. Therefore, these studies demonstrate that p38α is a major upstream activator of MAPKAP kinase 2 and a key component of the IL-1 signaling pathway. However, p38α does not serve an indispensable role in apoptosis.

scholarly journals Adrenocortical function and regulation of the steroid 21-hydroxylase gene in NGFI-B-deficient mice.

1995 ◽  
Vol 15 (8) ◽  
pp. 4331-4316 ◽  
Author(s):  
P A Crawford ◽  
Y Sadovsky ◽  
K Woodson ◽  
S L Lee ◽  
J Milbrandt
Keyword(s):  
Early Gene ◽  
Adrenocortical Function ◽  
Mrna Levels ◽  
Orphan Nuclear Receptor ◽  
Wild Type ◽  
Gene Encoding ◽  
Serum Corticosterone ◽  
B Mice

The immediate-early gene NGFI-B encodes an orphan nuclear receptor that binds DNA as a monomer and activates transcription through a canonical response element (NBRE). NGFI-B is expressed under basal conditions and in response to external stimuli in many mammalian tissues. In particular, NGFI-B expression is dramatically elevated in the adrenal cortex in response to stress and in Y1 adrenocortical cells in response to adrenocorticotropin. NGFI-B activates transcription through an NBRE of the gene encoding 21-hydroxylase (P450c21) in Y1 cells. Steroidogenic factor 1 (SF-1), a homolog of NGFI-B, also activates the P450c21 promoter. To examine the influence of these factors on P450c21 expression in vivo and the function of the hypothalamic-pituitary-adrenocortical axis as a whole, we generated NGFI-B (-/-) mice. These mice thrive and reproduce normally and maintain normal basal adrenocorticotropin, corticosterone, and P450c21 mRNA levels. In response to increases in adrenocorticotropin, NGFI-B (-/-) and wild-type mice demonstrated equivalent increases in serum corticosterone levels. Furthermore, and in contrast to in vitro results, no increases in P450c21 mRNA levels were observed in response to increases in adrenocorticotropin in NGFI-B (-/-) or wild-type mice. While SF-1 mRNA levels were not increased with increased steroidogenic demand, adrenal expression of Nurr1, a close homolog of NGFI-B, was induced to a greater extent by lipopolysaccharide in NGFI-B (-/-) mice than in wild-type mice. Finally, when the administration of dexamethasone for suppression was stopped, P450c21 mRNA and serum corticosterone levels recovered at the same rate in wild-type and NGFI-B (-/-) mice. Thus, while NGFI-B appears poised to affect the structure and function of the adrenal gland, the gland functions normally in its absence, suggesting that other factors, including Nurr1 and SF-1, are sufficient to drive P450c21 expression in mice and maintain normal steroidogenesis.

scholarly journals Application of human iPS cell-derived vascular cell differentiation induction method to elucidation of vascular pathology

Impact ◽  
2021 ◽  
Vol 2021 (5) ◽  
pp. 16-18
Author(s):  
Daisuke Taura
Keyword(s):  
Endothelial Cells ◽  
Cell Differentiation ◽  
Animal Models ◽  
Vascular Endothelial Cells ◽  
Es Cells ◽  
Ips Cells ◽  
Vascular Endothelial ◽  
Vascular Cell ◽  
Mural Cells ◽  
Human Ips Cells

Cardiovascular diseases such as arteriosclerosis are a leading cause of death in developed countries. Various factors, including diet, exercise and genetics, can play a role in the onset of such conditions and the development of treatment options is complex and often involves the use of animal models. However, there are limitations with the use of animal models due to their inherent differences to humans, meaning that research results aren't always translatable. Associate Professor Daisuke Taura, Department of Diabetes, Endocrinology and Nutrition, Kyoto University, Japan, is interested in the use of in vitro techniques for investigating cardiovascular conditions. He is conducting vascular cell differentiation induction research using human embryonic stem cells (ES) cells and induced pluripotent cells (iPS), which can be induced to differentiate into any type of human cell, provided the right culturing conditions are present. In a world first, Taura successfully induced vascular constituent cells from human iPS cells, which led to important results in the establishment of vascular cells with autosomal dominant polycystic kidney disease (ADPKD) and moyamoya disease. Ultimately, Taura and his team are working towards a radical treatment for atherosclerotic diseases, and are also seeking to induce differentiation of vascular constituent cells using human ES/ iPS cells and explore their development and differentiation processes. Taura has been successful in improving on culture methods and, in doing so, achieved the differentiation of human ES cells into vascular endothelial cells and mural cells, and the differentiation of human iPS cells into vascular endothelial cells and mural cells.

Vascular endothelial cell lineage-specific promoter in transgenic mice

Development ◽  
1995 ◽  
Vol 121 (4) ◽  
pp. 1089-1098 ◽  
Author(s):  
T.M. Schlaeger ◽  
Y. Qin ◽  
Y. Fujiwara ◽  
J. Magram ◽  
T.N. Sato
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Vascular Endothelial Cells ◽  
Es Cells ◽  
Vascular Endothelial Cell ◽  
Cell Lineage ◽  
Vascular Endothelial ◽  
Lacz Reporter

Vascular endothelial cells play essential roles in the function and development of the cardiovascular system. However, due to the lack of lineage-specific markers suitable for molecular and biochemical analyses, very little is known about the molecular mechanisms that regulate endothelial cell differentiation. We report the first vascular endothelial cell lineage-specific (including angioblastic precursor cells) 1.2 kb promoter in transgenic mice. Moreover, deletion analysis of this promoter region in transgenic embryos revealed multiple elements that are required for the maximum endothelial cell lineage-specific expression. This is a powerful molecular tool that will enable us to identify factors and cellular signals essential for the establishment of vascular endothelial cell lineage. It will also allow us to deliver genes specifically into this cell type in vivo to test specifically molecules that have been implicated in cardiovascular development. Furthermore, we have established embryonic stem (ES) cells from the blastocysts of the transgenic mouse that carry the 1.2 kb promoter-LacZ reporter transgene. These ES cells were able to differentiate in vitro to form cystic embryoid bodies (CEB) that contain endothelial cells determined by PECAM immunohistochemistry. However, these in vitro differentiated endothelial cells did not express the LacZ reporter gene. This indicates the lack of factors and/or cellular interactions which are required to induce the expression of the reporter gene mediated by this 1.2 kb promoter in this in vitro differentiation system. Thus this system will allow us to screen for the putative inducers that exist in vivo but not in vitro. These putative inducers are presumably important for in vivo differentiation of vascular endothelial cells.

scholarly journals Formation of Methyl Mercaptan froml-Methionine by Porphyromonas gingivalis

2000 ◽  
Vol 68 (12) ◽  
pp. 6912-6916 ◽  
Author(s):  
Mamiko Yoshimura ◽  
Yoshio Nakano ◽  
Yoshihisa Yamashita ◽  
Takahiko Oho ◽  
Toshiyuki Saito ◽  
...  
Keyword(s):  
Porphyromonas Gingivalis ◽  
Wild Type Strain ◽  
Marked Decrease ◽  
Oral Bacteria ◽  
Methyl Mercaptan ◽  
Wild Type ◽  
Oral Malodor ◽  
Gene Encoding ◽  
Degradation Activity

ABSTRACT Methyl mercaptan production by oral bacteria is thought to be one of the main causes of oral malodor. We examined the ability of periodontopathic Porphyromonas gingivalis to produce methyl mercaptan from l-methionine and found that the invasive strains W83 and W50 produced large amounts of methyl mercaptan. We cloned and sequenced the mgl gene encodingl-methionine-α-deamino-γ-mercaptomethane-lyase (METase) from P. gingivalis W83. The structural mgl gene consisted of 1,200 bp and encoded a 43.3-kDa protein. To examine the role of methyl mercaptan in the pathogenesis ofP. gingivalis, a METase-deficient mutant of P. gingivalis W83 was constructed. The methionine degradation activity and virulence of the mutant (M1217) and the parent strain (W83) in mice were compared. M1217 showed a marked decrease in the formation of methyl mercaptan from l-methionine and decreased virulence compared with the wild-type strain W83. These results suggest that methyl mercaptan not only is one of the sources of oral malodor, but may also play a role in the pathogenicity of P. gingivalis.

scholarly journals Platelet adhesion to podoplanin under flow is mediated by the receptor CLEC-2 and stabilised by Src/Syk-dependent platelet signalling

2015 ◽  
Vol 113 (05) ◽  
pp. 1109-1120 ◽  
Author(s):  
Alice Y. Pollitt ◽  
Kate Lowe ◽  
Arusa Latif ◽  
Gerard B. Nash ◽  
Steve P. Watson ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Platelet Adhesion ◽  
Vascular Endothelial Cells ◽  
Vascular Endothelial ◽  
Mouse Development ◽  
Shear Rates ◽  
Flow Perfusion ◽  
Platelet Binding ◽  
Lymphatic Development ◽  
The Stability

SummaryPlatelet-specific deletion of CLEC-2, which signals through Src and Syk kinases, or global deletion of its ligand podoplanin results in bloodfilled lymphatics during mouse development. Platelet-specific Syk deficiency phenocopies this defect, indicating that platelet activation is required for lymphatic development. In the present study, we investigated whether CLEC-2-podoplanin interactions could support platelet arrest from blood flow and whether platelet signalling is required for stable platelet adhesion to lymphatic endothelial cells (LECs) and recombinant podoplanin under flow. Perfusion of human or mouse blood over human LEC monolayers led to platelet adhesion and aggregation. Following α∥bβ3 blockade, individual platelets still adhered. Platelet binding occurred at venous but not arterial shear rates. There was no adhesion using CLEC-2-deficient blood or to vascular endothelial cells (which lack podoplanin). Perfusion of human blood over human Fc-podoplanin (hFcPDPN) in the presence of monoclonal antibody IV.3 to block FcγR∥A receptors led to platelet arrest at similar shear rates to those used on LECs. Src and Syk inhibitors significantly reduced global adhesion of human or mouse platelets to LECs and hFcPDPN. A similar result was seen using Syk-deficient mouse platelets. Reduced platelet adhesion was due to a decrease in the stability of binding. In conclusion, our data reveal that CLEC-2 is an adhesive receptor that supports platelet arrest to podoplanin under venous shear. Src/Syk-dependent signalling stabilises platelet adhesion to podoplanin, providing a possible molecular mechanism contributing to the lymphatic defects of Syk-deficient mice.

Thrombomodulin-mediated catabolism of protein C by pleural mesothelial and vascular endothelial cells

2007 ◽  
Vol 98 (09) ◽  
pp. 627-634 ◽  
Author(s):  
Alireza Rezaie ◽  
Steven Idell ◽  
Alexei Iakhiaev
Keyword(s):  
Endothelial Cells ◽  
Protein C ◽  
Vascular Endothelial Cells ◽  
Degradation Products ◽  
Cell Types ◽  
Model Systems ◽  
Vascular Endothelial ◽  
Wild Type ◽  
Endothelial Protein C Receptor ◽  
Gla Domain

SummaryPleural mesothelial and vascular endothelial cells express protein C (PC) pathway components including thrombomodulin (TM) and endothelial protein C receptor (EPCR) and activate PC by the thrombin-TM dependent mechanism.We used these cells as model systems to identify molecules involved in endocytosis and degradation of PC. We find that mesothelial and endothelial cells can bind, internalize and degrade PC.Addition of thrombin markedly induced degradation of PC by these cells in a TM-dependent fashion, implicating the involvement of the thrombin-TM complex in internalization and degradation of PC. This observation defines a novel function for the thrombin-TM complex as a degradation receptor for PC and suggests that PC is degraded concurrent with its activation.A PC Gla-domain mutant, which is unable to bind to the EPCR, was degraded by the cells to a lesser extent than wild-type PC, implicating the PC degradation concurrent with its activation. Consistent with the role of thrombin-TM complex as a degradation receptor, the catalytically inactive thrombin-S195A also induced PC degradation though to a lesser extent than wild-type thrombin.This suggests that generation of activated PC (APC) can contribute to accumulation of degradation products, but is not essential for the thrombin-induced degradation of PC. The thrombin-TMmediated degradation of PC by both cell types suggest a previously unrecognized mechanism, which can contribute to PC consumption.This mechanism may be pathophysiologically relevant and can contribute to an acquired PC deficiency in conditions characterized by sustained thrombin generation.

scholarly journals Human Cytomegalovirus UL47 Tegument Protein Functions after Entry and before Immediate-Early Gene Expression

2002 ◽  
Vol 76 (3) ◽  
pp. 1043-1050 ◽  
Author(s):  
Jill T. Bechtel ◽  
Thomas Shenk
Keyword(s):  
Human Cytomegalovirus ◽  
Human Fibroblasts ◽  
Open Reading Frame ◽  
Early Gene ◽  
Immediate Early ◽  
Wild Type Virus ◽  
Marker Genes ◽  
Type Virus ◽  
Wild Type ◽  
Reading Frame

ABSTRACT The human cytomegalovirus UL47 open reading frame encodes a 110-kDa protein that is a component of the virion tegument. We have constructed a cytomegalovirus mutant, ADsubUL47, in which the central portion of the UL47 open reading frame has been replaced by two marker genes. The mutant replicated to titers 100-fold lower than those for wild-type virus after infection at either a high or a low input multiplicity in primary human fibroblasts but was substantially complemented on cells expressing UL47 protein. A revertant virus in which the mutation was repaired, ADrevUL47, replicated with wild-type kinetics. Mutant virions lacked UL47 protein and contained reduced amounts of UL48 protein. The mutant was found to be less infectious than wild-type virus, and a defect very early in the replication cycle was observed. Transcription of the viral immediate-early 1 gene was delayed by 8 to 10 h. However, this delay was not the result of a defect in virus entry or of the inability of virion proteins to transactivate the major immediate-early promoter. We also show that the UL47 protein coprecipitated with the UL48 and UL69 tegument proteins and the UL86-encoded major capsid protein. We propose that a UL47-containing complex is involved in the release of viral DNA from the disassembling virus particle and that the loss of UL47 protein causes this process to be delayed.

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