The Myosin II ATPase Inhibitor Blebbistatin Prevents Thrombin-Induced Inhibition of Intercellular Calcium Wave Propagation in Corneal Endothelial Cells

2008 ◽  
Vol 49 (11) ◽  
pp. 4816 ◽  
Author(s):  
Raf Ponsaerts ◽  
Catheleyne D'hondt ◽  
Geert Bultynck ◽  
Sangly P. Srinivas ◽  
Johan Vereecke ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Wave Propagation ◽  
Myosin Ii ◽  
Calcium Wave ◽  
Corneal Endothelial Cells ◽  
Atpase Inhibitor

scholarly journals Thrombin Inhibits Intercellular Calcium Wave Propagation in Corneal Endothelial Cells by Modulation of Hemichannels and Gap Junctions

2007 ◽  
Vol 48 (1) ◽  
pp. 120 ◽  
Author(s):  
Catheleyne D’hondt ◽  
Raf Ponsaerts ◽  
Sangly P. Srinivas ◽  
Johan Vereecke ◽  
Bernard Himpens
Keyword(s):  
Endothelial Cells ◽  
Wave Propagation ◽  
Gap Junctions ◽  
Calcium Wave ◽  
Corneal Endothelial Cells

Adenosine induces dephosphorylation of myosin II regulatory light chain in cultured bovine corneal endothelial cells

2004 ◽  
Vol 79 (4) ◽  
pp. 543-551 ◽  
Author(s):  
S.P. Srinivas ◽  
M. Satpathy ◽  
P. Gallagher ◽  
E. Larivière ◽  
W. Van Driessche
Keyword(s):  
Endothelial Cells ◽  
Light Chain ◽  
Myosin Ii ◽  
Regulatory Light Chain ◽  
Corneal Endothelial Cells

Endothelial cell retraction is induced by PAK2 monophosphorylation of myosin II

2000 ◽  
Vol 113 (3) ◽  
pp. 471-482 ◽  
Author(s):  
Q. Zeng ◽  
D. Lagunoff ◽  
R. Masaracchia ◽  
Z. Goeckeler ◽  
G. Cote ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Actin Cytoskeleton ◽  
Light Chain ◽  
Myosin Ii ◽  
Specific Inhibitor ◽  
Regulatory Light Chain ◽  
Atpase Inhibitor ◽  
Cytoskeletal Rearrangements ◽  
Cell Retraction

The p21-activated kinase (PAK) family includes several enzyme isoforms regulated by the GTPases Rac1 and Cdc42. PAK1, found in brain, muscle and spleen, has been implicated in triggering cytoskeletal rearrangements such as the dissolution of stress fibers and reorganization of focal complexes. The role of the more widely distributed PAK2 in controlling the cytoskeleton has been less well studied. Previous work has demonstrated that PAK2 can monophosphorylate the myosin II regulatory light chain and induce retraction of permeabilized endothelial cells. In this report we characterize PAK2's morphological and biochemical effect on intact endothelial cells utilizing microinjection of constitutively active PAK2. Under these conditions we observed a modification of the actin cytoskeleton with retraction of endothelial cell margins accompanied by an increase in monophosphorylation of myosin II. Selective inhibitors were used to analyze the mechanism of action of PAK2. Staurosporine, a direct inhibitor of PAK2, largely prevented the action of microinjected PAK2 in endothelial cells. Butanedione monoxime, a non-specific myosin ATPase inhibitor, also inhibited the effects of PAK2 implicating myosin in the changes in cytoskeletal reorganization. In contrast, KT5926, a specific inhibitor of myosin light chain kinase was ineffective in preventing the changes in morphology and the actin cytoskeleton. The additional finding that endogenous PAK2 associates with myosin II is consistent with the proposal that cell retraction and cytoskeletal rearrangements induced by microinjected PAK2 depend on the direct activation of myosin II by PAK2 monophosphorylation of the regulatory light chain.

scholarly journals Calcium Wave Propagation in Networks of Endothelial Cells: Model-based Theoretical and Experimental Study

2012 ◽  
Vol 8 (12) ◽  
pp. e1002847 ◽  
Author(s):  
Juexuan Long ◽  
Michael Junkin ◽  
Pak Kin Wong ◽  
James Hoying ◽  
Pierre Deymier
Keyword(s):  
Experimental Study ◽  
Endothelial Cells ◽  
Wave Propagation ◽  
Calcium Wave ◽  
Model Based

scholarly journals Connexin 40 and ATP-dependent intercellular calcium wave in renal glomerular endothelial cells

2008 ◽  
Vol 294 (6) ◽  
pp. R1769-R1776 ◽  
Author(s):  
Ildikó Toma ◽  
Eric Bansal ◽  
Elliott J. Meer ◽  
Jung Julie Kang ◽  
Sarah L. Vargas ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Wave Propagation ◽  
Fold Increase ◽  
Extracellular Atp ◽  
Physical Contact ◽  
Calcium Wave ◽  
Fluorescence Confocal Microscopy ◽  
Glomerular Endothelial Cells ◽  
Connexin 40 ◽  
Voltage Dependent

Endothelial intracellular calcium ([Ca2+]i) plays an important role in the function of the juxtaglomerular vasculature. The present studies aimed to identify the existence and molecular elements of an endothelial calcium wave in cultured glomerular endothelial cells (GENC). GENCs on glass coverslips were loaded with Fluo-4/Fura red, and ratiometric [Ca2+]iimaging was performed using fluorescence confocal microscopy. Mechanical stimulation of a single GENC caused a nine-fold increase in [Ca2+]i, which propagated from cell to cell throughout the monolayer (7.9 ± 0.3 μm/s) in a regenerative manner (without decrement of amplitude, kinetics, and speed) over distances >400 μm. Inhibition of voltage-dependent calcium channels with nifedipine had no effect on the above parameters, but the removal of extracellular calcium reduced Δ[Ca2+]iby 50%. Importantly, the gap junction uncoupler α-glycyrrhetinic acid or knockdown of connexin 40 (Cx40) by transfecting GENCs with Cx40 short interfering RNA (siRNA) almost completely eliminated Δ[Ca2+]iand the calcium wave. Breakdown of extracellular ATP using a scavenger cocktail (apyrase and hexokinase) or nonselective inhibition of purinergic P2 receptors with suramin, had similar blocking effects. Scraping cells off along a line eliminated physical contact between cells but did not effect calcium wave propagation. Using an ATP biosensor technique, we detected a significant elevation in extracellular ATP (Δ = 76 ± 2 μM) during calcium wave propagation, which was abolished by Cx40 siRNA treatment (Δ = 6 ± 1 μM). These studies suggest that connexin 40 hemichannels and extracellular ATP are key molecular elements of the glomerular endothelial calcium wave, which may serve important juxtaglomerular functions.

scholarly journals Adenosine Opposes Thrombin-Induced Inhibition of Intercellular Calcium Wave in Corneal Endothelial Cells

2007 ◽  
Vol 48 (4) ◽  
pp. 1518 ◽  
Author(s):  
Catheleyne D’hondt ◽  
Sangly P. Srinivas ◽  
Johan Vereecke ◽  
Bernard Himpens
Keyword(s):  
Endothelial Cells ◽  
Calcium Wave ◽  
Corneal Endothelial Cells
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