ZO-1 Redistribution and F-actin stress fiber formation in pulmonary endothelial cells after thermal injury

Joseph T. Murphy, Steve Duffy, Ronald G. Tompkins

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

Background In response to isolated inflammatory stimuli, changes in endothelial cell morphology that enhance paracellular flow of solutes result from F-actin stress fiber formation, myosin phosphorylation, and actin anchoring protein (ZO-1) modifications. We hypothesized that myosin light chain kinase inhibition would diminish burn-enhanced endothelial monolayer permeability by secondarily preventing F-actin and actin anchoring protein rearrangements. Methods Human pulmonary microvascular endothelial cells were treated for 4 hours with 20% human burn serum (isolated from patients with > 45% total body surface area thermal injury or healthy volunteers). Select cultures were pretreated with myosin light chain kinase inhibitors (ML-9). Permeability was assessed by migration of bovine serum albumin across cell monolayers. Cells were stained with rhodamine-phalloidin and anti-ZO-1 antisera and examined by means of confocal microscopy. Results Burn serum significantly enhanced monolayer permeability to albumin, whereas pretreatment with ML-9 limited this effect. Control cells maintained cortical F-actin and peripheral ZO-1 distributions (1a, b), whereas burn serum induced transcellular F-actin stress fiber formation and a diffuse ZO-1 staining (2a, b). ML-9 prevented burn-induced actin rearrangements, but not the diffuse redistribution of ZO-1. Conclusion These data demonstrate that endothelial F-actin stress fiber formation and ZO-1 redistribution contribute to postburn loss of pulmonary endothelial monolayer integrity. Although myosin phosphorylation appears to be required for endothelial F-actin stress fiber formation, redistribution of actin-membrane anchoring proteins appears to be regulated independently after thermal injury.

Original languageEnglish (US)
Pages (from-to)81-90
Number of pages10
JournalJournal of Trauma
Volume54
Issue number1
DOIs
StatePublished - Jan 2003

Keywords

  • Cytoskeleton
  • Endothelium
  • MLCK
  • Permeability
  • Thermal injury
  • ZO-1

ASJC Scopus subject areas

  • Surgery
  • Critical Care and Intensive Care Medicine

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