Endothelial myosin light chain kinase regulates neutrophil migration across human umbilical vein endothelial cell monolayer.

Although extravasation of neutrophils is a critical step in acute inflammation, the role of the endothelial cytoskeleton in neutrophil transmigration has not been fully investigated. We used an in vitro model of neutrophil transmigration across a monolayer of HUVEC cultured on amniotic membrane. Human neutrophils were allowed to migrate across the HUVEC monolayer in response to a gradient leukotriene B4 and then the number of migrated neutrophils were counted microscopically. We also followed endothelial F-actin and myosin filament formation using rhodamine-phalloidin and anti-myosin Ab staining. Myosin light chain (MLC) phosphorylation in endothelial cells was determined by immunoprecipitation of 32P-labeled HUVEC with anti-myosin polyclonal Ab. Normally, neutrophil migration induced F-actin formation, myosin filament formation, and MLC phosphorylation in HUVEC. When HUVEC was pretreated with the myosin light chain kinase (MLCK) inhibitor, ML-9, neutrophil migration was diminished and F-actin formation, myosin filament formation, and MLC phosphorylation were inhibited. Pretreatments of HUVEC with the intracellular calcium ion chelator, bis-(O-aminophenoxyl)ethane-N, N, N', N'-tetraacetic acid acetoxymethyl ester (BAPTA/AM), and the calmodulin antagonist, trifluoperazine, had similar effects. These results indicate that a calcium/ calmodulin-dependent MLCK in endothelial cells regulates neutrophil transendothelial migration.

[1]  S I Simon,et al.  Neutrophil transendothelial migration is independent of tight junctions and occurs preferentially at tricellular corners. , 1997, Journal of immunology.

[2]  F. Luscinskas,et al.  Endothelial-dependent Mechanisms Regulate Leukocyte Transmigration: A Process Involving the Proteasome and Disruption of the Vascular Endothelial–Cadherin Complex at Endothelial Cell-to-Cell Junctions , 1997, The Journal of experimental medicine.

[3]  Yoshiharu Matsuura,et al.  Phosphorylation and Activation of Myosin by Rho-associated Kinase (Rho-kinase)* , 1996, The Journal of Biological Chemistry.

[4]  Kozo Kaibuchi,et al.  Regulation of Myosin Phosphatase by Rho and Rho-Associated Kinase (Rho-Kinase) , 1996, Science.

[5]  G. Schmid-Schönbein,et al.  Transvascular and Interstitial Migration of Neutrophils in Rat Mesentery , 1996, Microcirculation.

[6]  M. Lampugnani,et al.  Thrombin-induced increase in endothelial permeability is associated with changes in cell-to-cell junction organization. , 1996, Arteriosclerosis, thrombosis, and vascular biology.

[7]  J. Bodmer,et al.  Histamine and thrombin modulate endothelial focal adhesion through centripetal and centrifugal forces. , 1996, The Journal of clinical investigation.

[8]  D L Rimm,et al.  Alpha 1(E)-catenin is an actin-binding and -bundling protein mediating the attachment of F-actin to the membrane adhesion complex. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[9]  R B Wysolmerski,et al.  Myosin light chain kinase-regulated endothelial cell contraction: the relationship between isometric tension, actin polymerization, and myosin phosphorylation , 1995, The Journal of cell biology.

[10]  K. Kurokawa,et al.  Involvement of rho in GTPγS‐induced enhancement of phosphorylation of 20 kDa myosin light chain in vascular smooth muscle cells: inhibition of phosphatase activity , 1995, FEBS letters.

[11]  A. P. Soler,et al.  Interaction of alpha-actinin with the cadherin/catenin cell-cell adhesion complex via alpha-catenin , 1995, The Journal of cell biology.

[12]  A. Verin,et al.  Regulation of endothelial cell gap formation and barrier function by myosin-associated phosphatase activities. , 1995, The American journal of physiology.

[13]  J. Sun,et al.  IFN-gamma and TNF-alpha induce redistribution of PECAM-1 (CD31) on human endothelial cells. , 1995, Journal of immunology.

[14]  H. Davis,et al.  Regulation of endothelial cell gap formation and barrier dysfunction: Role of myosin light chain phosphorylation , 1995, Journal of cellular physiology.

[15]  J. Gamble,et al.  Transendothelial migration of neutrophils involves integrin-associated protein (CD47). , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[16]  D. Walker,et al.  The structure of the tricellular region of endothelial tight junctions of pulmonary capillaries analyzed by freeze-fracture. , 1994, Microvascular research.

[17]  C. Smith,et al.  Adhesion molecules and inflammatory injury , 1994, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[18]  W. Muller,et al.  Monoclonal antibody to murine PECAM-1 (CD31) blocks acute inflammation in vivo , 1994, The Journal of experimental medicine.

[19]  S. Thom,et al.  Involvement of platelet-endothelial cell adhesion molecule-1 in neutrophil recruitment in vivo. , 1993, Science.

[20]  R. Sheldon,et al.  Role of myosin light-chain phosphorylation in endothelial cell retraction. , 1993, The American journal of physiology.

[21]  P. Kubes,et al.  Effects of human neutrophil elastase (HNE) on neutrophil function in vitro and in inflamed microvessels. , 1993, Blood.

[22]  A. Moy,et al.  The effect of histamine and cyclic adenosine monophosphate on myosin light chain phosphorylation in human umbilical vein endothelial cells. , 1993, The Journal of clinical investigation.

[23]  S. Silverstein,et al.  Endothelial cell cytosolic free calcium regulates neutrophil migration across monolayers of endothelial cells , 1993, The Journal of cell biology.

[24]  J. D. Pardee,et al.  Actin-facilitated assembly of smooth muscle myosin induces formation of actomyosin fibrils , 1992, The Journal of cell biology.

[25]  C. Benjamin,et al.  E-selectin (endothelial-leukocyte adhesion molecule-1) is not required for the migration of neutrophils across IL-1-stimulated endothelium in vitro. , 1992, Journal of immunology.

[26]  M. Itoh,et al.  A 220-kD undercoat-constitutive protein: its specific localization at cadherin-based cell-cell adhesion sites , 1991, The Journal of cell biology.

[27]  T. Kuijpers,et al.  Neutrophil and monocyte adherence to and migration across monolayers of cytokine-activated endothelial cells: the contribution of CD18, ELAM-1, and VLA-4. , 1991, Blood.

[28]  C W Smith,et al.  Monoclonal antibodies to leukocyte integrins CD11a/CD18 and CD11b/CD18 or intercellular adhesion molecule-1 inhibit chemoattractant-stimulated neutrophil transendothelial migration in vitro. , 1991, Blood.

[29]  W. Gerthoffer.  Regulation of the contractile element of airway smooth muscle. , 1991, The American journal of physiology.

[30]  R. Wysolmerski,et al.  Regulation of permeabilized endothelial cell retraction by myosin phosphorylation. , 1991, American Journal of Physiology.

[31]  L. McIntire,et al.  Chemotactic factors regulate lectin adhesion molecule 1 (LECAM-1)-dependent neutrophil adhesion to cytokine-stimulated endothelial cells in vitro. , 1991, The Journal of clinical investigation.

[32]  J. Sellers,et al.  Substance P contracts bovine tracheal smooth muscle via activation of myosin light chain kinase. , 1990, The American journal of physiology.

[33]  M B Furie,et al.  Migration of neutrophils across endothelial monolayers is stimulated by treatment of the monolayers with interleukin-1 or tumor necrosis factor-alpha. , 1989, Journal of immunology.

[34]  K. Vaughan,et al.  Actin filaments mediate Dictyostelium myosin assembly in vitro. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[35]  R. Adelstein,et al.  Antigen-induced secretion of histamine and the phosphorylation of myosin by protein kinase C in rat basophilic leukemia cells. , 1989, The Journal of biological chemistry.

[36]  M. Kazatchkine,et al.  Platelet‐dependent induction and amplification of polymorphonuclear leucocyte lysosomal enzyme release , 1989, British journal of haematology.

[37]  D. Hartshorne,et al.  Correlation of enzymatic properties and conformation of bovine erythrocyte myosin. , 1989, Biochemistry.

[38]  C W Smith,et al.  Recognition of an endothelial determinant for CD 18-dependent human neutrophil adherence and transendothelial migration. , 1988, The Journal of clinical investigation.

[39]  Clive R. Bagshaw,et al.  Active site trapping of nucleotide by smooth and non-muscle myosins. , 1988, Journal of molecular biology.

[40]  L. Liebovitch,et al.  Effects of human neutrophil chemotaxis across human endothelial cell monolayers on the permeability of these monolayers to ions and macromolecules , 1988, Journal of cellular physiology.

[41]  J. Koretz,et al.  Effects of phosphorylation of light chain residues threonine 18 and serine 19 on the properties and conformation of smooth muscle myosin. , 1988, The Journal of biological chemistry.

[42]  T. A. Sutton,et al.  Phosphorylation of two sites on smooth muscle myosin. Effects on contraction of glycerinated vascular smooth muscle. , 1988, The Journal of biological chemistry.

[43]  H. Hidaka,et al.  Selective inhibition of catalytic activity of smooth muscle myosin light chain kinase. , 1987, The Journal of biological chemistry.

[44]  K. Weber,et al.  Calcium control of actin-myosin based contraction in triton models of mouse 3T3 fibroblasts is mediated by the myosin light chain kinase (MLCK)-calmodulin complex. , 1983, Experimental cell research.

[45]  E. Jaffe,et al.  Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. , 1973, The Journal of clinical investigation.

[46]  R. Wysolmerski,et al.  Involvement of myosin light-chain kinase in endothelial cell retraction. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[47]  R. A. Murphy,et al.  Ca2+, crossbridge phosphorylation, and contraction. , 1989, Annual review of physiology.

[48]  R. Ezzell,et al.  Evidence for regulation of lamellipodial and tail contraction of glycerinated chicken embryonic fibroblasts by myosin light chain kinase. , 1986, Cell motility and the cytoskeleton.

[49]  J. Kendrick‐Jones,et al.  Light-chain phosphorylation controls the conformation of vertebrate non-muscle and smooth muscle myosin molecules , 1983, Nature.