title: Role of N-WASP in Endothelial Monolayer Formation and Integrity.
abstract: Endothelial cells (ECs) form a monolayer that serves as a barrier between the blood and the underlying tissue. ECs tightly regulate their cell-cell junctions, controlling the passage of soluble materials and immune cells across the monolayer barrier. We studied the role of N-WASP, a key regulator of Arp2/3 complex and actin assembly, in EC monolayers. We report that N-WASP regulates endothelial monolayer integrity by affecting the organization of cell junctions. Depletion of N-WASP resulted in an increase in transendothelial electrical resistance, a measure of monolayer integrity. N-WASP depletion increased the width of cell-cell junctions and altered the organization of F-actin and VE-cadherin at junctions. N-WASP was not present at cell-cell junctions in monolayers under resting conditions, but it was recruited following treatment with sphingosine-1-phosphate. Taken together, our results reveal a novel role for N-WASP in remodeling EC junctions, which is critical for monolayer integrity and function.
authors.affiliation: From the Department of Cell Biology and Physiology, Washington University, St. Louis, Missouri 63110.
authors.forename: Olivia L
authors.initials: OL
authors.lastname: Mooren
authors.affiliation: From the Department of Cell Biology and Physiology, Washington University, St. Louis, Missouri 63110.
authors.forename: Joanna
authors.initials: J
authors.lastname: Kim
authors.affiliation: From the Department of Cell Biology and Physiology, Washington University, St. Louis, Missouri 63110.
authors.forename: Jinmei
authors.initials: J
authors.lastname: Li
authors.affiliation: From the Department of Cell Biology and Physiology, Washington University, St. Louis, Missouri 63110 jacooper@wustl.edu.
authors.forename: John A
authors.initials: JA
authors.lastname: Cooper
mesh_terms: D051376:Actin-Related Protein 2-3 Complex / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D015703:Antigens, CD / Q000378:metabolism; D015820:Cadherins / Q000378:metabolism; D002460:Cell Line; D003599:Cytoskeleton / Q000378:metabolism*; D042783:Endothelial Cells / Q000378:metabolism*; D006801:Humans; D007365:Intercellular Junctions / Q000378:metabolism*; D008246:Lysophospholipids / Q000378:metabolism; D034741:RNA, Small Interfering; D012867:Skin / Q000166:cytology / Q000378:metabolism; D013110:Sphingosine / Q000031:analogs & derivatives / Q000378:metabolism; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal / Q000235:genetics / Q000378:metabolism*; D000099152:Cadherin 5
chemical_list: D051376:Actin-Related Protein 2-3 Complex; D000199:Actins; D015703:Antigens, CD; D015820:Cadherins; D008246:Lysophospholipids; D034741:RNA, Small Interfering; D013110:Sphingosine; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal; D000099152:Cadherin 5; C495771:WASL protein, human; C060506:sphingosine 1-phosphate
keywords: Arp2/3 complex; N-WASP; actin; barrier; cytoskeleton; endothelial cell; endothelial dysfunction; monolayer