title: F-actin-rich contractile endothelial pores prevent vascular leakage during leukocyte diapedesis through local RhoA signalling.
abstract: During immune surveillance and inflammation, leukocytes exit the vasculature through transient openings in the endothelium without causing plasma leakage. However, the exact mechanisms behind this intriguing phenomenon are still unknown. Here we report that maintenance of endothelial barrier integrity during leukocyte diapedesis requires local endothelial RhoA cycling. Endothelial RhoA depletion in vitro or Rho inhibition in vivo provokes neutrophil-induced vascular leakage that manifests during the physical movement of neutrophils through the endothelial layer. Local RhoA activation initiates the formation of contractile F-actin structures that surround emigrating neutrophils. These structures that surround neutrophil-induced endothelial pores prevent plasma leakage through actomyosin-based pore confinement. Mechanistically, we found that the initiation of RhoA activity involves ICAM-1 and the Rho GEFs Ect2 and LARG. In addition, regulation of actomyosin-based endothelial pore confinement involves ROCK2b, but not ROCK1. Thus, endothelial cells assemble RhoA-controlled contractile F-actin structures around endothelial pores that prevent vascular leakage during leukocyte extravasation.
authors.affiliation: Department of Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Academic Medical Centre, University of Amsterdam, 1066CX Amsterdam, The Netherlands.
authors.forename: Niels
authors.initials: N
authors.lastname: Heemskerk
authors.affiliation: Department of Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Academic Medical Centre, University of Amsterdam, 1066CX Amsterdam, The Netherlands.
authors.forename: Lilian
authors.initials: L
authors.lastname: Schimmel
authors.affiliation: Department of Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Academic Medical Centre, University of Amsterdam, 1066CX Amsterdam, The Netherlands.
authors.forename: Chantal
authors.initials: C
authors.lastname: Oort
authors.affiliation: Department of Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Academic Medical Centre, University of Amsterdam, 1066CX Amsterdam, The Netherlands.
authors.forename: Jos
authors.initials: J
authors.lastname: van Rijssel
authors.affiliation: Genetics and Developmental Biology, Center for Cell Analyses and Modelling, University of Connecticut Health Centre, Farmington, Connecticut 06032, USA.
authors.forename: Taofei
authors.initials: T
authors.lastname: Yin
authors.affiliation: Experimental Medicine and Pharmacology, Centre for Microvascular Research, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary, University of London, Charterhouse Square, London, EC1M 6BQ, UK.
authors.forename: Bin
authors.initials: B
authors.lastname: Ma
authors.affiliation: Molecular Cytology, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam 1098XH, The Netherlands.
authors.forename: Jakobus
authors.initials: J
authors.lastname: van Unen
authors.affiliation: Department of Angiogenesis, Walter-Brendel-Center of Experimental Medicine Ludwig-Maximilians University Marchioninistr. 27 81377 Munich, Germany.
authors.forename: Bettina
authors.initials: B
authors.lastname: Pitter
authors.affiliation: Department of Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Academic Medical Centre, University of Amsterdam, 1066CX Amsterdam, The Netherlands.
authors.forename: Stephan
authors.identifier: http://orcid.org/0000-0002-1091-475X
authors.initials: S
authors.lastname: Huveneers
authors.affiliation: Molecular Cytology, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam 1098XH, The Netherlands.
authors.forename: Joachim
authors.identifier: http://orcid.org/0000-0002-0630-3825
authors.initials: J
authors.lastname: Goedhart
authors.affiliation: Genetics and Developmental Biology, Center for Cell Analyses and Modelling, University of Connecticut Health Centre, Farmington, Connecticut 06032, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Angiogenesis, Walter-Brendel-Center of Experimental Medicine Ludwig-Maximilians University Marchioninistr. 27 81377 Munich, Germany.
authors.forename: Eloi
authors.initials: E
authors.lastname: Montanez
authors.affiliation: Experimental Medicine and Pharmacology, Centre for Microvascular Research, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary, University of London, Charterhouse Square, London, EC1M 6BQ, UK.
authors.forename: Abigail
authors.initials: A
authors.lastname: Woodfin
authors.affiliation: Department of Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Academic Medical Centre, University of Amsterdam, 1066CX Amsterdam, The Netherlands.
authors.forename: Jaap D
authors.initials: JD
authors.lastname: van Buul
mesh_terms: D008841:Actin Cytoskeleton / Q000737:chemistry / Q000378:metabolism; D000199:Actins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002199:Capillary Permeability; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D018799:Intercellular Adhesion Molecule-1; D007962:Leukocytes / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009504:Neutrophils / Q000166:cytology / Q000378:metabolism; D015398:Signal Transduction; D057705:Transendothelial and Transepithelial Migration; D020742:rhoA GTP-Binding Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: D000199:Actins; D018799:Intercellular Adhesion Molecule-1; D020742:rhoA GTP-Binding Protein
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