scholarly journals Association of hyaluronan with rat vascular endothelial and smooth muscle cells.

1995 ◽  
Vol 43 (7) ◽  
pp. 689-697 ◽  
Author(s):  
P S Eggli ◽  
W Graber
Keyword(s):  
Smooth Muscle ◽  
Plasma Membranes ◽  
Vascular Tissue ◽  
Cell Types ◽  
Vascular Endothelial ◽  
Cell Plasma ◽  
One Step ◽  
Lowicryl K4m ◽  
20 Nm ◽  
Hyaluronan Binding

We investigated the distribution of hyaluronan (hyaluronic acid) in rat vascular tissue fixed by an osmium tetroxide (or glutaraldehyde) microwave technique and embedded in epoxy resin (or Lowicryl K4M), using hyaluronan binding proteins coupled to 15-20-nm gold particles as ultrastructural markers in a one-step post-embedding procedure. The intra- and extracellular aspects of vascular endothelial and smooth muscle cell plasma membranes revealed distinct labeling, with a high affinity for caveolae being manifested in both cell types and in all kinds of vessels. Hyaluronan was also localized intercellularly in areas characterized by extensive endothelial cell interdigitation. Intracellularly, moderate staining of nuclear heterochromatin was observed.

Regulation Of Vascular Prostacyclin (PGI2) Synthesis

1981 ◽  
Author(s):  
S Coughlin ◽  
M Moskowitz ◽  
H N Antoniades ◽  
L Levine
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Cell Types ◽  
Feedback Mechanism ◽  
Vascular Endothelial ◽  
Serum Factor ◽  
Negative Feedback Mechanism ◽  
Cells In Culture ◽  
Other Substances

We have examined the possibility that substances released during platelet degranulation modify vascular PGI2 synthesis. PGI2 is a potent inhibitor of platelet function produced by vascular endothelial and smooth muscle cells. Regulation of PGI2 synthesis by blood vessels is not well understood. We report that a platelet- dependent factor in serum dramatically stimulates PGI2 synthesis by vascular endothelial and smooth muscle cells in culture. We further report that platelet-derived growth factor (PDGF), a releasable protein found in platelet alpha granules, stimulates PGI2 synthesis by the above cell types by over 100 fold. The concentration of PDGF required to elicit this effect is below that reported in human serum. The above mentioned serum factor is relatively heat stable, non-dialyzable, and cationic; preliminary studies indicate that anti-PDGF antiserum is capable of blocking stimulation of PGI2 synthesis by both PDGF and serum. These data suggest that the serum factor may indeed be PDGF. PDGF acts synergistically with other platelet granule constituents (serotonin, ATP) and with thrombin to stimulate PGI2 synthesis by vascular cells in culture. We thus postulate that platelet-released PDGF, in concert with other substances generated during clotting, acts to increase vessel wall PGI2 synthesis as part of a negative feedback mechanism controlling platelet aggregation. A defect in the ability of a blood vessel to increase PGI2 production in response to platelet degranulation, as may occur in atherosclerotic vessels, could perhaps contribute to the genesis of thromboembolic events.

A molecular basis for synexin-driven, calcium-dependent membrane fusion

1988 ◽  
Vol 139 (1) ◽  
pp. 267-286
Author(s):  
H. B. Pollard ◽  
A. L. Burns ◽  
E. Rojas
Keyword(s):  
Membrane Fusion ◽  
Calcium Binding ◽  
Plasma Membranes ◽  
Cell Types ◽  
Complete Analysis ◽  
Calcium Dependent ◽  
Cell Plasma ◽  
Probable Role ◽  
Highly Hydrophobic ◽  
Chromaffin Granule Ghosts

Membranes of secretory vesicles fuse with each other and with plasma membranes during exocytosis in many different cell types. The probable role of calcium in the process is now widely accepted, and it is possible that at least one cytosolic mediator of calcium action is synexin. Synexin is a 47,000 Mr calcium-binding protein, initially discovered in the bovine adrenal medulla, which binds to granule membranes and to inner aspects of chromaffin cell plasma membranes. Synexin causes chromaffin granules to aggregate, and such aggregates can be caused to fuse in the additional presence of arachidonic acid. Synexin also mediates the direct fusion of liposomes and chromaffin granule ghosts. To understand better the mechanisms of membrane fusion promoted by synexin we have attempted to define the primary sequence of the protein. Our initial efforts were directed towards purification of bovine synexin in sufficient amounts to allow us to sequence tryptic peptides. However, as the project progressed we also directed our attention to human synexin, preparing peptides from this protein as well. From analysis of bovine peptides we learned that the synexin molecule might be closely related to a class of proteins including lipocortin I, calpactin (p36), endonexin II, protein II and calelectrin 67K. Complete analysis of a human synexin cDNA clone revealed strong homology with bovine synexin. The analysis also showed that synexin contained a unique, long, highly hydrophobic N-terminal leader sequence followed by a characteristic four-fold repeat homologous with those found in other members of the synexin gene family. The highly hydrophobic character of synexin seems consistent with information previously obtained that synexin is able to insert directly into the interior of bilayers prepared not only from purified phosphatidylserine but also from biological membranes. The evidence for such insertions is a dramatic increase in the capacitance of the membrane, formed at the tip of a patch pipette, when calcium-activated synexin is applied to the bilayer. Additional evidence is the fact that synexin also forms calcium-selective channels when the protein is applied to the cytosolic aspect of the plasmalemma when that side is also exposed to calcium at sub-millimolar concentrations. Thus, the synexin molecule not only enters the membrane, but also spans it. From these and other data we have developed the concept that the fusion process may involve synexin forming a ‘hydrophobic bridge’ between two fusing membranes. Lipid movement across this bridge may then be the material basis for final fusion.(ABSTRACT TRUNCATED AT 400 WORDS)

Cell type specific electrophoretic polypeptide profiles of cultured bovine cells

1984 ◽  
Vol 72 (1) ◽  
pp. 135-145
Author(s):  
D.W. Lincoln ◽  
K.I. Braunschweiger ◽  
W.R. Braunschweiger ◽  
J.R. Smith
Keyword(s):  
Smooth Muscle ◽  
Pulmonary Artery ◽  
Smooth Muscle Cells ◽  
Vascular Endothelial Cells ◽  
Muscle Cells ◽  
Cell Types ◽  
Vascular Endothelial ◽  
Molecular Weights ◽  
Cell Type Specific ◽  
Electrophoretic Techniques

The polypeptide profiles of bovine vascular endothelial cells (from pulmonary artery and descending aorta), smooth muscle cells (from pulmonary artery) and fibroblast cells (from skin and lung) were examined by high-resolution two-dimensional polyacrylamide gel electrophoretic techniques. A set of polypeptides (molecular weights between 43 X 10(3) and 47 X 10(3) and pI values from 6.0-4.8, respectively) exhibited patterns that were unique to the three cell types. In the case of smooth muscle cells, these polypeptides exhibited cell-density-dependent expression. These results allow for identification of the three cell types on the basis of their highly specific polypeptide signatures.

scholarly journals Physical contact between human vascular endothelial and smooth muscle cells modulates cytosolic and nuclear calcium homeostasis

2018 ◽  
Vol 96 (7) ◽  
pp. 655-661
Author(s):  
Ghada S. Hassan ◽  
Danielle Jacques ◽  
Pedro D’Orléans-Juste ◽  
Sheldon Magder ◽  
Ghassan Bkaily
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Culture Medium ◽  
Vascular Endothelial Cells ◽  
Muscle Cells ◽  
Cell Types ◽  
Physical Contact ◽  
Vascular Endothelial ◽  
Nuclear Calcium ◽  
Calcium Dependent

The interaction between vascular endothelial cells (VECs) and vascular smooth muscle cells (VSMCs) plays an important role in the modulation of vascular tone. There is, however, no information on whether direct physical communication regulates the intracellular calcium levels of human VECs (hVECs) and (or) human VSMCs (hVSMCs). Thus, the objective of the study is to verify whether co-culture of hVECs and hVSMCs modulates cytosolic ([Ca2+]c) and nuclear calcium ([Ca2+]n) levels via physical contact and (or) factors released by both cell types. Quantitative 3D confocal microscopy for [Ca2+]c and [Ca2+]n measurement was performed in cultured hVECs or hVSMCs or in co-culture of hVECs–hVSMCs. Our results show that: (1) physical contact between hVECs–hVECs or hVSMCs–hVSMCs does not affect [Ca2+]c and [Ca2+]n in these 2 cell types; (2) physical contact between hVECs and hVSMCs induces a significant increase only of [Ca2+]n of hVECs without affecting the level of [Ca2+]c and [Ca2+]n of hVSMCs; and (3) preconditioned culture medium of hVECs or hVSMCs does not affect [Ca2+]c and [Ca2+]n of both types of cells. We concluded that physical contact between hVECs and hVSMCs only modulates [Ca2+]n in hVECs. The increase of [Ca2+]n in hVECs may modulate nuclear functions that are calcium dependent.

Progesterone increases nitric oxide synthesis in human vascular endothelial cells through activation of membrane progesterone receptor-α

2015 ◽  
Vol 308 (10) ◽  
pp. E899-E911 ◽  
Author(s):  
Yefei Pang ◽  
Jing Dong ◽  
Peter Thomas
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Progesterone Receptor ◽  
Map Kinase ◽  
Plasma Membranes ◽  
Vascular Endothelial Cells ◽  
No Production ◽  
Vascular Endothelial ◽  
Enos Phosphorylation ◽  
20 Nm

Progesterone exerts beneficial effects on the human cardiovascular system by inducing rapid increases in nitric oxide (NO) production in vascular endothelial cells, but the receptors mediating these nongenomic progesterone actions remain unclear. Using human umbilical vein endothelial cells (HUVECs) as a model, we show that progesterone binds to plasma membranes of HUVECs with the characteristics of membrane progesterone receptors (mPRs). The selective mPR agonist Org OD 02-0 had high binding affinity for the progesterone receptor on HUVEC membranes, whereas nuclear PR (nPR) agonists R5020 and medroxyprogesterone acetate displayed low binding affinities. Immunocytochemical and Western blot analyses confirmed that mPRs are expressed in HUVECs and are localized on their plasma membranes. NO levels increased rapidly after treatment with 20 nM progesterone, Org OD 02-0, and a progesterone-BSA conjugate but not with R5020, suggesting that this progesterone action is at the cell surface and initiated through mPRs. Progesterone and Org OD 02-0 (20 nM) also significantly increased endothelial nitric oxide synthase (eNOS) activity and eNOS phosphorylation. Knockdown of mPRα expression by treatment with small-interfering RNA (siRNA) blocked the stimulatory effects of 20 nM progesterone on NO production and eNOS phosphorylation, whereas knockdown of nPR was ineffective. Treatment with PI3K/Akt and MAP kinase inhibitors blocked the stimulatory effects of progesterone, Org OD 02-0, and progesterone-BSA on NO production and eNOS phosphorylation and also prevented progesterone- and Org OD 02-0-induced increases in Akt and ERK phosphorylation. The results suggest that progesterone stimulation of NO production in HUVECs is mediated by mPRα and involves signaling through PI3K/Akt and MAP kinase pathways.

scholarly journals Metabolism of adenine nucleotides by ectoenzymes of vascular endothelial and smooth-muscle cells in culture

1980 ◽  
Vol 190 (2) ◽  
pp. 421-429 ◽  
Author(s):  
J D Pearson ◽  
J S Carleton ◽  
J L Gordon
Keyword(s):  
Smooth Muscle ◽  
Substrate Specificity ◽  
Smooth Muscle Cells ◽  
Adenine Nucleotides ◽  
Nucleoside Diphosphate Kinase ◽  
Muscle Cells ◽  
Blood Platelet ◽  
Cell Types ◽  
Vascular Endothelial ◽  
Cells In Culture

1. Pig aortic endothelial and smooth-muscle cells in culture rapidly catabolize exogenous ATP, ADP or AMP. 2. In both cell types catabolism is due to Mg2+-stimulated ectoenzymes. 3. Inhibition and substrate-specificity studies suggest that both cell types possess three distinct ectonucleotidases, namely nucleoside triphosphatase (EC 3.6.1.15), nucleoside diphosphatase (EC 3.6.1.6) and 5′-nucleotidase (EC 3.1.3.5), as well as nucleoside diphosphate kinase (EC 2.7.4.6). 4. These ectonucleotidase systems could be of importance in the regulation of neurotransmission, blood platelet function and vasodilation.

Whole-cell and nuclear NADPH oxidases levels and distribution in human endocardial endothelial, vascular smooth muscle, and vascular endothelial cells

2013 ◽  
Vol 91 (1) ◽  
pp. 71-79 ◽  
Author(s):  
Lena Ahmarani ◽  
Levon Avedanian ◽  
Johny Al-Khoury ◽  
Claudine Perreault ◽  
Danielle Jacques ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Vascular Endothelial Cells ◽  
Vascular Diseases ◽  
Cell Types ◽  
Ros Generation ◽  
Vascular Endothelial ◽  
Whole Cell ◽  
Cardiovascular Cells ◽  
Nuclear Levels

The results of our study show that whole-cell and nuclear levels of NADPH oxidase-1 (NOX1) are similar in human vascular endothelial cells (hVECs) and smooth muscle cells (hVSMCs), but lower in human endocardial endothelial cells (hEECs). NOX2 levels were higher in hVECs and lower in hVSMCs. NOX3 levels were the same in hVECs and hVSMCs, but lower in hEECs. NOX4 levels were similar in all of the cell types. NOX4 levels were higher in hVECs than in hVSMCs. NOX5 was also present throughout the 3 cell types, including their nuclei, in the following order: hEECs > hVSMCs > hVECs. The level of basal reactive oxygen species (ROS) was highest in hVECs and lowest in hVSMCs. However, the Ca2+ level was highest in hVSMCs and lowest in hVECs. These findings suggest that all types of NOXs exist in hEECs, hVECs, and hVSMCs, although their density and distribution are cell-type dependent. The density of the different NOXs correlated with the ROS level, but not with the Ca2+ level. In conclusion, NOXs, including NOX3, exist in cardiovascular cells and their nuclei. The nucleus is a major source of ROS generation. The nuclear NOXs may contribute to ROS and Ca2+ homeostasis, which may affect cell remodeling, including the formation of nuclear T-tubules in vascular diseases and aging.

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