Advertisement

Direct observation and quantitative analysis of spatiotemporal dynamics of individual living monocytes during transendothelial migration

  • Ken Hashimoto
    Correspondence
    Corresponding author. Tel.: +81 86 462 1111x83512; fax: +81 86 464 1020.
    Affiliations
    Department of Physiology, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama 701-0192, Japan

    Department of Cardiovascular Physiology and Medical Engineering, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama 700-8558, Japan
    Search for articles by this author
  • Noriyuki Kataoka
    Affiliations
    Department of Medical Engineering, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama 701-0192, Japan

    Department of Cardiovascular Physiology and Medical Engineering, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama 700-8558, Japan
    Search for articles by this author
  • Emi Nakamura
    Affiliations
    Department of Physiology, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama 701-0192, Japan
    Search for articles by this author
  • Hiroko Asahara
    Affiliations
    Department of Physiology, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama 701-0192, Japan
    Search for articles by this author
  • Yasuo Ogasawara
    Affiliations
    Department of Medical Engineering, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama 701-0192, Japan
    Search for articles by this author
  • Katsuhiko Tsujioka
    Affiliations
    Department of Physiology, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama 701-0192, Japan
    Search for articles by this author
  • Fumihiko Kajiya
    Affiliations
    Department of Medical Engineering, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama 701-0192, Japan

    Department of Cardiovascular Physiology and Medical Engineering, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama 700-8558, Japan
    Search for articles by this author

      Abstract

      Objective:

      To visualize and quantitatively analyze spatiotemporal dynamics of individual living monocytes during transendothelial migration (TEM).

      Methods and results:

      We developed an in vitro new experimental system using confocal laser scanning microscope with following two improvements: (1) ultra thin collagen gel layer (30–50 μm thick) constructed under human umbilical vein endothelial cell layer for three-dimensional observation with high magnification; (2) appropriate fluorescent labeling of living monocytes and endothelial cells to keep highest cell activity. Individual monocytes behaved quite diversely. Approximately 70% of adhered monocytes directionally crawled to intercellular junction, and started invasion. Time from adhesion to start of invasion was 8.6 ± 5.4 min (mean ± S.D., n = 61 monocytes). Approximately 80% of such invading monocytes completed TEM, but remaining 20% of once invading monocytes hesitated transmigration, and returned onto the endothelial surface. Time from start to finish of invasion was 6.3 ± 3.2 min (mean ± S.D., n = 53 monocytes).

      Conclusions:

      Using our collagen gel-based newly-developed system, we visualized and quantitatively analyzed detailed spatiotemporal, three-dimensional dynamics of individual living monocytes during TEM. We revealed that monocytes encountered at least two hurdles, at starting invasion, and leaving endothelium, to achieve complete TEM. Approximately 56% (80% of 70% of adhered monocytes) passed both hurdles.

      Keywords

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Atherosclerosis
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Libby P.
        • Ridker P.M.
        • Maseri A.
        Inflammation and atherosclerosis.
        Circulation. 2002; 105: 1135-1143
        • Allport J.R.
        • Muller W.A.
        • Luscinskas F.W.
        Monocytes induce reversible focal changes in vascular endothelial cadherin complex during transendothelial migration under flow.
        J. Cell Biol. 2000; 148: 203-216
        • Berman M.E.
        • Xie Y.
        • Muller W.A.
        Roles of platelet/endothelial cell adhesion molecule-1 (PECAM-1, CD31) in natural killer cell transendothelial migration and beta 2 integrin activation.
        J. Immunol. 1996; 156: 1515-1524
        • Cooper D.
        • Lindberg F.P.
        • Gamble J.R.
        • Brown E.J.
        • Vadas M.A.
        Transendothelial migration of neutrophils involves integrin-associated protein (CD47).
        Proc. Natl. Acad Sci. USA. 1995; 92: 3978-3982
        • Mamdouh Z.
        • Chen X.
        • Pierini L.M.
        • Maxfield F.R.
        • Muller W.A.
        Targeted recycling of PEC AM from endothelial surface-connected compartments during diapedesis.
        Nature. 2003; 421: 748-753
        • Sans E.
        • Delachanal E.
        • Duperray A.
        Analysis of the roles of ICAM-1 in neutrophil transmigration using a reconstituted mammalian cell expression model: implication of ICAM-1 cytoplasmic domain and Rho-dependent signaling pathway.
        J. Immunol. 2001; 166: 544-551
        • Schenkel A.R.
        • Mamdouh Z.
        • Chen X.
        • Liebman R.M.
        • Muller W.A.
        CD99 plays a major role in the migration of monocytes through endothelial junctions.
        Nat. Immunol. 2002; 3: 143-150
        • Strey A.
        • Janning A.
        • Barth H.
        • Gerke V.
        Endothelial Rho signaling is required for monocyte transendothelial migration.
        FEBS Lett. 2002; 517: 261-266
        • Sultana C.
        • Shen Y.
        • Rattan V.
        • Kalra V.K.
        Lipoxygenase metabolites induced expression of adhesion molecules and transendothelial migration of monocyte-like HL-60 cells is linked to protein kinase C activation.
        J. Cell Physiol. 1996; 167: 477-487
        • Iwaki K.
        • Ohashi K.
        • Ikeda M.
        • et al.
        Decrease in the amount of focal adhesion kinase (p125(FAK)) in interleukin-1beta-stimulated human umbilical vein endothelial cells by binding of human monocytic cell lines.
        J. Biol. Chem. 1997; 272: 20665-20670
        • Giaever I.
        • Keese C.R.
        Micromotion of mammalian cells measured electrically.
        Proc. Natl. Acad Sci. USA. 1991; 88: 7896-7900
        • Kataoka N.
        • Iwaki K.
        • Hashimoto K.
        • et al.
        Measurements of endothelial cell-to-cell and cell-to-substrate gaps and micromechanical properties of endothelial cells during monocyte adhesion.
        Proc. Natl. Acad Sci. USA. 2002; 99: 15638-15643
        • Bamforth S.D.
        • Lightman S.L.
        • Greenwood J.
        Ultrastructural analysis of interleukin-1 beta-induced leukocyte recruitment to the rat retina.
        Invest Ophthalmol Vis Sci. 1997; 38: 25-35
        • Feng D.
        • Nagy J.A.
        • Pyne K.
        • Dvorak H.F.
        • Dvorak A.M.
        Neutrophils emigrate from venules by a transendothelial cell pathway in response to FMLP.
        J. Exp. Med. 1998; 187: 903-915
        • Hoshi O.
        • Ushiki T.
        Scanning electron microscopic studies on the route of neutrophil extravasation in the mouse after exposure to the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP).
        Arch. Histol. Cytol. 1999; 62: 253-260
        • Miyoshi M.
        • Mizutani T.
        • Iizuka A.
        Migration of granular leukocytes through the perikaryal portion of endothelial cytoplasm: electron microscope observation in the mouse gallbladder vessels.
        Kaibogaku Zasshi Japanese. 1970; 45: 217-224
        • Faustmann P.M.
        • Dermietzel R.
        Extravasation of polymorphonuclear leukocytes from the cerebral microvasculature. Inflammatory response induced by alpha-bungarotoxin.
        Cell Tissue Res. 1985; 242: 399-407
        • Takaku M.
        • Wada Y.
        • Jinnouchi K.
        • et al.
        An in vitro coculture model of transmigrant monocytes and foam cell formation.
        Arterioscler Thromb Vasc Biol. 1999; 19: 2330-2339
        • Burns A.R.
        • Bowden R.A.
        • MacDonell S.D.
        • et al.
        Analysis of tight junctions during neutrophil transendothelial migration.
        J. Cell Sci. 2000; 113: 45-57
        • Burns A.R.
        • Walker D.C.
        • Brown E.S.
        • et al.
        Neutrophil transendothelial migration is independent of tight junctions and occurs preferentially at tricellular corners.
        J. Immunol. 1997; 159: 2893-2903
        • Kitayama J.
        • Hidemura A.
        • Saito H.
        • Nagawa H.
        Shear stress affects migration behavior of polymorphonuclear cells arrested on endothelium.
        Cell Immunol. 2000; 203: 39-46
        • Migliorisi G.
        • Folkes E.
        • Pawlowski N.
        • Cramer E.B.
        In vitro studies of human monocyte migration across endothelium in response to leukotriene B4 and f-Met-Leu-Phe.
        Am. J. Pathol. 1987; 127: 157-167
        • Giri R.
        • Shen Y.
        • Stins M.
        • et al.
        beta-amyloid-induced migration of monocytes across human brain endothelial cells involves RAGE and PECAM-1.
        Am. J. Physiol. Cell Physiol. 2000; 279: C1772-C1781
        • Worthylake R.A.
        • Lemoine S.
        • Watson J.M.
        • Burridge K.
        RhoA is required for monocyte tail retraction during transendothelial migration.
        J. Cell Biol. 2001; 154: 147-160
        • Brown E.B.
        • Campbell R.B.
        • Tsuzuki Y.
        • et al.
        In vivo measurement of gene expression, angiogenesis and physiological function in tumors using multiphoton laser scanning microscopy.
        Nat. Med. 2001; 7 ([Erratum in: Nat. Med. 2001;7:1069]): 864-868
        • Friedl P.
        • Brocker E.B.
        Reconstructing leukocyte migration in 3D extracellular matrix by time-lapse videomicroscopy and computer-assisted tracking.
        Methods Mol. Biol. 2004; 239: 77-90
        • Denk W.
        • Strickler J.H.
        • Webb W.W.
        Two-photon laser scanning fluorescence microscopy.
        Science. 1990; 248: 73-76
        • Buchstaller A.
        • Kunz S.
        • Berger P.
        • et al.
        Cell adhesion molecules NgCAM and axonin-1 form heterodimers in the neuronal membrane and cooperate in neurite outgrowth promotion.
        J. Cell Biol. 1996; 135: 1593-1607
        • Hu Y.L.
        • Li S.
        • Miao H.
        • et al.
        Roles of microtubule dynamics and small GTPase Rac in endothelial cell migration and lamellipodium formation under flow.
        J. Vasc Res. 2002; 39: 465-476