Analysis of tight junctions during neutrophil transendothelial migration

Alan R. Burns, Robert A. Bowden, Sonia D. MacDonell, David C. Walker, Timothy O. Odebunmi, Elizabeth M. Donnachie, Scott I. Simon, Mark L. Entman, C. Wayne Smith

Research output: Contribution to journalArticle

155 Citations (Scopus)

Abstract

Intercellular junctions have long been considered the main sites through which adherent neutrophils (PMNs) penetrate the endothelium. Tight junctions (TJs; zonula occludens) are the most apical component of the intercellular cleft and they form circumferential belt-like regions of intimate contact between adjacent endothelial cells. Whether PMN transmigration involves disruption of the TJ complex is unknown. We report here that endothelial TJs appear to remain intact during PMN adhesion and transmigration. Human umbilical vein endothelial cell (HUVEC) monolayers, a commonly used model for studying leukocyte trafficking, were cultured in astrocyte-conditioned medium to enhance TJ expression. Immunofluorescence microscopy and immunoblot analysis showed that activated PMN adhesion to resting monolayers or PMN migration across interleukin-1-treated monolayers does not result in widespread proteolytic loss of TJ proteins (ZO-1, ZO-2, and occludin) from endothelial borders. Ultrastructurally, TJs appear intact during and immediately following PMN transendothelial migration. Similarly, transendothelial electrical resistance is unaffected by PMN adhesion and migration. Previously, we showed that TJs are inherently discontinuous at tricellular corners where the borders of three endothelial cells meet and PMNs migrate preferentially at tricellular corners. Collectively, these results suggest that PMN migration at tricellular corners preserves the barrier properties of the endothelium and does not involve widespread disruption of endothelial TJs.

Original languageEnglish (US)
Pages (from-to)45-57
Number of pages13
JournalJournal of Cell Science
Volume113
Issue number1
StatePublished - 2000
Externally publishedYes

Fingerprint

Transendothelial and Transepithelial Migration
Tight Junctions
Endothelium
Neutrophils
Endothelial Cells
Occludin
Intercellular Junctions
Human Umbilical Vein Endothelial Cells
Conditioned Culture Medium
Electric Impedance
Interleukin-1
Fluorescence Microscopy
Astrocytes
Leukocytes
Proteins

Keywords

  • Endothelium
  • Inflammation
  • Neutrophil
  • Tight junction

ASJC Scopus subject areas

  • Cell Biology

Cite this

Burns, A. R., Bowden, R. A., MacDonell, S. D., Walker, D. C., Odebunmi, T. O., Donnachie, E. M., ... Smith, C. W. (2000). Analysis of tight junctions during neutrophil transendothelial migration. Journal of Cell Science, 113(1), 45-57.

Analysis of tight junctions during neutrophil transendothelial migration. / Burns, Alan R.; Bowden, Robert A.; MacDonell, Sonia D.; Walker, David C.; Odebunmi, Timothy O.; Donnachie, Elizabeth M.; Simon, Scott I.; Entman, Mark L.; Smith, C. Wayne.

In: Journal of Cell Science, Vol. 113, No. 1, 2000, p. 45-57.

Research output: Contribution to journalArticle

Burns, AR, Bowden, RA, MacDonell, SD, Walker, DC, Odebunmi, TO, Donnachie, EM, Simon, SI, Entman, ML & Smith, CW 2000, 'Analysis of tight junctions during neutrophil transendothelial migration', Journal of Cell Science, vol. 113, no. 1, pp. 45-57.
Burns AR, Bowden RA, MacDonell SD, Walker DC, Odebunmi TO, Donnachie EM et al. Analysis of tight junctions during neutrophil transendothelial migration. Journal of Cell Science. 2000;113(1):45-57.
Burns, Alan R. ; Bowden, Robert A. ; MacDonell, Sonia D. ; Walker, David C. ; Odebunmi, Timothy O. ; Donnachie, Elizabeth M. ; Simon, Scott I. ; Entman, Mark L. ; Smith, C. Wayne. / Analysis of tight junctions during neutrophil transendothelial migration. In: Journal of Cell Science. 2000 ; Vol. 113, No. 1. pp. 45-57.
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