Regulation of endothelial barrier function during flow-induced conversion to an arterial phenotype.
OBJECTIVE
Flow-induced conversion of endothelial cells into an elongated arterial phenotype requires a coordinated regulation of cell junctions. Here we investigated the effect of acute and chronic flow on junction regulation.


RESULTS
Using an extended experimental setup that allows analyses of endothelial barrier function under flow conditions, we found a flow-induced upregulation of the transendothelial electrical resistance within minutes. This was accompanied by an increase in actin filaments along the junctions and vascular endothelial (VE)-cadherin clustering, which was identified at nanoscale resolution by stimulated emission depletion microscopy. In addition, a transient tyrosine phosphorylation of VE-cadherin and catenins occurred within minutes following the onset of flow. VE-cadherin and actin distribution were maintained under chronic flow over 24 h and associated with the upregulation of VE-cadherin and alpha-catenin expression, thus compensating for the cell elongation-mediated increase in cell border length. Importantly, all observed effects were rac1 dependent as verified by the inhibitory effect of dominant negative N17rac1.


CONCLUSIONS
These results show that flow-induced conversion of endothelial cells into an arterial phenotype occurs while intercellular junctions remain intact. The data place rac1 in a central multimodal regulatory position that might be important in the development of vascular diseases, such as arteriosclerosis.
Seebach
Jochen
J
Institute of Physiology, Medical Faculty Dresden of the TU-Dresden, 01307 Dresden, Germany.
Donnert
Gerald
G
Kronstein
Romy
R
Werth
Sebastian
S
Wojciak-Stothard
Beata
B
Falzarano
Darryl
D
Mrowietz
Christof
C
Hell
Stefan W
SW
Schnittler
Hans-J
HJ
D000199:Actins / Q000378:metabolism; D015703:Antigens, CD / Q000378:metabolism; D001158:Arteries; D015820:Cadherins / Q000378:metabolism; D002448:Cell Adhesion; D002462:Cell Membrane / Q000378:metabolism; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D004594:Electrophysiology; D042783:Endothelial Cells / Q000378:metabolism*; D006801:Humans; D007365:Intercellular Junctions / Q000378:metabolism*; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D010641:Phenotype; D010766:Phosphorylation; D012039:Regional Blood Flow; D013314:Stress, Mechanical; D014680:Veins; D020830:rac1 GTP-Binding Protein / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000378:metabolism; D000099152:Cadherin 5
D000199:Actins; D015703:Antigens, CD; D015820:Cadherins; D020830:rac1 GTP-Binding Protein; D020741:rho GTP-Binding Proteins; D000099152:Cadherin 5; C107953:RAC1 protein, human