Analysis of tight junctions during neutrophil transendothelial migration

• DOI: 10.1242/jcs.113.1.45
• Publication Date: 2021-04-25T17:31:54Z
• Authors: Alan R. Burns; Robert A. Bowden; Sonia D. MacDonell; David C. Walker; Timothy O. Odebunmi; Elizabeth M. Donn...; Scott I. Simon; Mark L. Entman; C. Wayne Smith
• Keywords: 0301 basic medicine; Hot Temperature; Neutrophils; Interleukins; Electric Conductivity; Membrane Proteins; Phosphoproteins; Neutrophil Activation; Tight Junctions; 3. Good health; Microscopy, Electron; 03 medical and health sciences; Microscopy, Fluorescence; Cell Movement; Culture Media, Conditioned; Occludin; Endopeptidases; Cell Adhesion; Freeze Fracturing; Humans; Endothelium, Vascular; Platelet Activating Factor; Cells, Cultured

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