scholarly journals MAGI1 Mediates eNOS Activation and NO Production in Endothelial Cells in Response to Fluid Shear Stress

Cells ◽  
2019 ◽  
Vol 8 (5) ◽  
pp. 388 ◽  
Author(s):  
Kedar Ghimire ◽  
Jelena Zaric ◽  
Begoña Alday-Parejo ◽  
Jochen Seebach ◽  
Mélanie Bousquenaud ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Shear Stress ◽  
Endothelial Cell ◽  
Protein Kinase ◽  
Fluid Shear Stress ◽  
Adaptor Protein ◽  
No Production ◽  
Fluid Shear ◽  
Enos Phosphorylation

Fluid shear stress stimulates endothelial nitric oxide synthase (eNOS) activation and nitric oxide (NO) production through multiple kinases, including protein kinase A (PKA), AMP-activated protein kinase (AMPK), AKT and Ca2+/calmodulin-dependent protein kinase II (CaMKII). Membrane-associated guanylate kinase (MAGUK) with inverted domain structure-1 (MAGI1) is an adaptor protein that stabilizes epithelial and endothelial cell-cell contacts. The aim of this study was to assess the unknown role of endothelial cell MAGI1 in response to fluid shear stress. We show constitutive expression and co-localization of MAGI1 with vascular endothelial cadherin (VE-cadherin) in endothelial cells at cellular junctions under static and laminar flow conditions. Fluid shear stress increases MAGI1 expression. MAGI1 silencing perturbed flow-dependent responses, specifically, Krüppel-like factor 4 (KLF4) expression, endothelial cell alignment, eNOS phosphorylation and NO production. MAGI1 overexpression had opposite effects and induced phosphorylation of PKA, AMPK, and CAMKII. Pharmacological inhibition of PKA and AMPK prevented MAGI1-mediated eNOS phosphorylation. Consistently, MAGI1 silencing and PKA inhibition suppressed the flow-induced NO production. Endothelial cell-specific transgenic expression of MAGI1 induced PKA and eNOS phosphorylation in vivo and increased NO production ex vivo in isolated endothelial cells. In conclusion, we have identified endothelial cell MAGI1 as a previously unrecognized mediator of fluid shear stress-induced and PKA/AMPK dependent eNOS activation and NO production.

Laminar Flow on Endothelial Cells Suppresses eNOS O-GlcNAcylation to Promote eNOS Activity

2021 ◽  
Author(s):  
Sarah Basehore ◽  
Samantha Bohlman ◽  
Callie Weber ◽  
Swathi Swaminathan ◽  
Yuji Zhang ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Shear Stress ◽  
Laminar Flow ◽  
No Production ◽  
Disturbed Flow ◽  
Enos Activity ◽  
Enos Phosphorylation ◽  
Steady Laminar ◽  
In Cells

Rationale: In diabetic animals as well as high glucose cell culture conditions, endothelial nitric oxide synthase (eNOS) is heavily O-GlcNAcylated, which inhibits its phosphorylation and nitric oxide (NO) production. It is unknown, however, whether varied blood flow conditions, which affect eNOS phosphorylation, modulate eNOS activity via O-GlcNAcylation-dependent mechanisms. Objective: The goal of this study was to test if steady laminar flow, but not oscillating disturbed flow, decreases eNOS O-GlcNAcylation, thereby elevating eNOS phosphorylation and NO production. Methods and Results: Human umbilical vein endothelial cells (HUVEC) were exposed to either laminar flow (20 dynes/cm2 shear stress) or oscillating disturbed flow (4{plus minus}6 dynes/cm2 shear stress) for 24 hours in a cone-and-plate device. eNOS O-GlcNAcylation was almost completely abolished in cells exposed to steady laminar but not oscillating disturbed flow. Interestingly, there was no change in protein level or activity of key O-GlcNAcylation enzymes (OGT, OGA, or GFAT). Instead, metabolomics data suggest that steady laminar flow decreases glycolysis and hexosamine biosynthetic pathway (HBP) activity, thereby reducing UDP-GlcNAc pool size and consequent O-GlcNAcylation. Inhibition of glycolysis via 2-deoxy-2-glucose (2-DG) in cells exposed to disturbed flow efficiently decreased eNOS O-GlcNAcylation, thereby increasing eNOS phosphorylation and NO production. Finally, we detected significantly higher O-GlcNAcylated proteins in endothelium of the inner aortic arch in mice, suggesting that disturbed flow increases protein O-GlcNAcylation in vivo. Conclusions: Our data demonstrate that steady laminar but not oscillating disturbed flow decreases eNOS O-GlcNAcylation by limiting glycolysis and UDP-GlcNAc substrate availability, thus enhancing eNOS phosphorylation and NO production. This research shows for the first time that O-GlcNAcylation is regulated by mechanical stimuli, relates flow-induced glycolytic reductions to macrovascular disease, and highlights targeting HBP metabolic enzymes in endothelial cells as a novel therapeutic strategy to restore eNOS activity and prevent EC dysfunction in cardiovascular disease.

Effects of Fluid Shear Stress on Endothelial Cell Invasion Into Three-Dimensional Matrices

2007 ◽  
Author(s):  
Hojin Kang ◽  
Kayla J. Bayless ◽  
Roland Kaunas
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Endothelial Cell ◽  
Cell Invasion ◽  
Fluid Shear Stress ◽  
Vascular Endothelial Cells ◽  
Umbilical Vein ◽  
Shear Stresses ◽  
Collagen Gels ◽  
Fluid Shear

We have previously developed a cell culture model to study the effects of angiogenic factors, such as sphingosine-1-phosphate (S1P), on the invasion of endothelial cells into the underlying extracellular matrix. In addition to biochemical stimuli, vascular endothelial cells are subjected to fluid shear stress due to blood flow. The present study is aimed at determining the effects of fluid shear stress on endothelial cell invasion into collagen gels. A device was constructed to apply well-defined fluid shear stresses to confluent human umbilical vein endothelial cells (HUVECs) seeded on collagen gels. Fluid shear stress induced significant increases in cell invasion with a maximal induction at ∼5 dyn/cm2. These results provide evidence that fluid shear stress is a significant stimulus for endothelial cell invasion and may play a role in regulating angiogenesis.

Glycated collagen alters endothelial cell actin alignment and nitric oxide release in response to fluid shear stress

2011 ◽  
Vol 44 (10) ◽  
pp. 1927-1935 ◽  
Author(s):  
Steven F. Kemeny ◽  
Dannielle S. Figueroa ◽  
Allison M. Andrews ◽  
Kenneth A. Barbee ◽  
Alisa Morss Clyne
Keyword(s):  
Nitric Oxide ◽  
Shear Stress ◽  
Endothelial Cell ◽  
Fluid Shear Stress ◽  
Fluid Shear ◽  
Nitric Oxide Release

Nitric oxide secretion by endothelial cells in response to fluid shear stress, aspirin, and temperature

2014 ◽  
Vol 103 (3) ◽  
pp. 1231-1237 ◽  
Author(s):  
Fatemeh Kabirian ◽  
Ghassem Amoabediny ◽  
Nooshin Haghighipour ◽  
Nasim Salehi-Nik ◽  
Behrouz Zandieh-Doulabi
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Shear Stress ◽  
Fluid Shear Stress ◽  
Fluid Shear

3-D Stress Analysis in Cultured Endothelial Cells Exposed to Fluid Shear Stress

1999 ◽  
Author(s):  
T. Ohashi ◽  
H. Sugawara ◽  
Y. Ishii ◽  
M. Sato
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Endothelial Cell ◽  
Cell Surface ◽  
Fluid Shear Stress ◽  
Flow Direction ◽  
Stress Distributions ◽  
Fluid Shear ◽  
Surface Geometry ◽  
Cultured Endothelial Cells

Abstract Under fluid shear stress, applied both in vivo and in vitro, vascular endothelial cells show morphological changes. After applying shear stress, cultured endothelial cells showed elongation and orientation to the flow direction (Kataoka et al., 1998). Moreover, statistical image analysis showed that intercellular F-actin distributions were confirmed to change depending on the shear stress and the flow direction. Thus, the endothelial cell morphology relates closely with the cytoskeletal structures. Intercellular stress distributions in the cells may be also accompanied by the reorganization of cytoskeletal structures. The use of both atomic force microscopy measurements (AFM) of endothelial cell surface topography and computational fluid dynamics of shear stress distributions acting on the cell surface, it has revealed that the surface geometry defined the detailed distribution of shear stress (Davies et al., 1995).

scholarly journals Fluid Shear Stress Stimulates Big Mitogen-activated Protein Kinase 1 (BMK1) Activity in Endothelial Cells

1999 ◽  
Vol 274 (1) ◽  
pp. 143-150 ◽  
Author(s):  
Chen Yan ◽  
Masafumi Takahashi ◽  
Masanori Okuda ◽  
Jiing-Dwan Lee ◽  
Bradford C. Berk
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Protein Kinase ◽  
Fluid Shear Stress ◽  
Mitogen Activated Protein Kinase ◽  
Fluid Shear ◽  
Mitogen Activated Protein

Abstract 1429: The Proline-Rich Tyrosine Kinase PYK2 Regulates NO Production by eNOS under Physiologic and Pathophysiologic Conditions.

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Annemarieke E Loot ◽  
Beate Fisslthaler ◽  
Rudi Busse ◽  
Ingrid Fleming
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Angiotensin Ii ◽  
Tyrosine Kinase ◽  
Carotid Arteries ◽  
Fluid Shear Stress ◽  
Dominant Negative ◽  
Maximal Response ◽  
No Production ◽  
Fluid Shear

In endothelial cells, the proline-rich tyrosine kinase (PYK2) is activated by fluid shear stress and phosphorylates the endothelial NO synthase (eNOS) on tyrosine residue 657. We have previously shown that eNOS phosphorylation on Tyr657 inhibits NO production in vitro and therefore addressed the consequences of PYK2 activation in a more physiological situation i.e., in perfused isolated carotid arteries as well as in response to other stimuli. Pressurised perfused murine carotid arteries subjected to step-wise increases in luminal flow responded with a graded endothelium-dependent vasodilatation that was partially sensitive to NOS inhibition. Carotid arteries in which the endothelium was infected with an adenovirus expressing a dominant-negative PYK2 mutant displayed normal flow-dependent vasodilatation (maximal response: R max 49 ± 6 μm) that was sensitive to the NOS inhibitor L-NAME (R max 28 ± 2 μm). However, flow-induced vasodilatation was markedly reduced in arteries overexpressing wild-type PYK2 and was insensitive to L-NAME (R max 27 ± 4 μm and 28 ± 5 μm, respectively) indicating loss of the NO-dependent component of vasodilatation. Given that angiotensin II is known to stimulate PYK2 phosphorylation and activation in smooth muscle cells we assessed the effects of angiotensin II on endothelial PYK2 activation and NO-dependent vasodilatation. We observed a time-dependent phosphorylation of PYK2 immuno-precipitated from porcine aortic endothelial cells stimulated with angiotensin II for 30 min up to 24 hrs. This correlated with an angiotensin II-induced decrease in NO-dependent vasodilatation. These data indicate that under physiological conditions, the tyrosine phosphorylation of eNOS by PYK2 in response to fluid shear stress attenuates the activity of the enzyme and limits NO-mediated flow-dependent vasodilatation. This PYK2-dependent inhibition of NO production may at least partially limit the detrimental consequences of maintained high NO output e.g. the generation of peroxynitrite. Conversely, under pathophysiological conditions in which angiotensin II levels are elevated, the increased activity of PYK2 can contribute to endothelial dysfunction by reducing eNOS activity.

Fluid shear stress-induced nitric oxide production in human cavernosal endothelial cells: inhibition by hyperglycaemia

2006 ◽  
Vol 97 (5) ◽  
pp. 1047-1052 ◽  
Author(s):  
HUNTER WESSELLS ◽  
THOMAS H. TEAL ◽  
KAREN ENGEL ◽  
CHRISTOPHER J. SULLIVAN ◽  
BYRON GALLIS ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Shear Stress ◽  
Fluid Shear Stress ◽  
Nitric Oxide Production ◽  
Fluid Shear ◽  
Oxide Production

scholarly journals Piezo1 acts upstream of TRPV4 to induce pathological changes in endothelial cells due to shear stress

2020 ◽  
pp. jbc.RA120.015059
Author(s):  
Sandip M Swain ◽  
Rodger A Liddle
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Endothelial Cell ◽  
Fluid Shear Stress ◽  
Venous Pressure ◽  
Calcium Signal ◽  
Cell Integrity ◽  
Fluid Shear ◽  
Channel Opening ◽  
Trpv4 Channel

The ion channels Piezo1 and TRPV4 have both, independently, been implicated in high venous pressure- and fluid shear stress-induced vascular hyperpermeability in endothelial cells.  However, the mechanism by which Piezo1 and TRPV4 channels execute the same function is poorly understood.  Here we demonstrate that Piezo1 regulates TRPV4 channel activation in endothelial cells and that Piezo1-mediated TRPV4 channel opening is a function of the strength and duration of fluid shear stress.  We first confirmed that either fluid shear stress or the Piezo1 agonist, Yoda1, led to an elevation in intracellular calcium ([Ca2+]i), and that application of the Piezo1 antagonist, GsMTx4, completely blocked this change. We discovered that high and prolonged shear stress caused sustained [Ca2+]i elevation which was blocked by inhibition of TRPV4 channel opening.  Moreover, Piezo1 stimulated TRPV4 opening through activation of phospholipase A2.  TRPV4-dependent sustained [Ca2+]i elevation was responsible for fluid shear stress- and Piezo1-mediated disruption of adherens junctions and actin remodeling.  Blockade of TRPV4 channels with the selective TRPV4 blocker, HC067047, prevented the loss of endothelial cell integrity and actin disruption induced by Yoda1 or shear stress and prevented Piezo1-induced monocyte adhesion to endothelial cell monolayers.  These findings demonstrate that Piezo1 activation by fluid shear stress initiates a calcium signal that causes TRPV4 opening which in turn is responsible for the sustained phase calcium elevation that triggers pathological events in endothelial cells.  Thus, deleterious effects of shear stress are initiated by Piezo1 but require TRPV4.

The Dynamic Response of Vascular Endothelial Cells to Fluid Shear Stress

1981 ◽  
Vol 103 (3) ◽  
pp. 177-185 ◽  
Author(s):  
C. F. Dewey ◽  
S. R. Bussolari ◽  
M. A. Gimbrone ◽  
P. F. Davies
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Endothelial Cell ◽  
Dynamic Response ◽  
Fluid Shear Stress ◽  
Vascular Endothelial Cells ◽  
Shear Stresses ◽  
Vascular Endothelial ◽  
Fluid Shear ◽  
Cell Functions

We have developed an in-vitro system for studying the dynamic response of vascular endothelial cells to controlled levels of fluid shear stress. Cultured monolayers of bovine aortic endothelial cells are placed in a cone-plate apparatus that produces a uniform fluid shear stress on replicate samples. Subconfluent endothelial cultures continuously exposed to 1–5 dynes/cm2 shear proliferate at a rate comparable to that of static cultures and reach the same saturation density (≃ 1.0–1.5 × 105 cells/cm2). When exposed to a laminar shear stress of 5–10 dynes/cm2, confluent monolayers undergo a time-dependent change in cell shape from polygonal to ellipsoidal and become uniformly oriented with flow. Regeneration of linear “wounds” in confluent monolayer appears to be influenced by the direction of the applied force. Preliminary studies indicate that certain endothelial cell functions, including fluid endocytosis, cytoskeletal assembly and nonthrombogenic surface properties, also are sensitive to shear stress. These observations suggest that fluid mechanical forces can directly influence endothelial cell structure and function. Modulation of endothelial behavior by fluid shear stresses may be relevant to normal vessel wall physiology, as well as the pathogenesis of vascular diseases, such as atherosclerosis.

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