Cyclic strain-mediated regulation of vascular endothelial occludin and ZO-1: influence on intercellular tight junction assembly and function

Nora T Collins, Philip M Cummins, Olga C Colgan, Gail Ferguson, Yvonne A Birney, Ronan P Murphy, Gerardene Meade, Paul A Cahill

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

OBJECTIVE: The vascular endothelium constitutes a highly effective fluid/solute barrier through the regulated apposition of intercellular tight junction complexes. Because endothelium-mediated functions and pathology are driven by hemodynamic forces (cyclic strain and shear stress), we hypothesized a dynamic regulatory link between endothelial tight junction assembly/function and hemodynamic stimuli. We, therefore, examined the effects of cyclic strain on the expression, modification, and function of 2 pivotal endothelial tight junction components, occludin and ZO-1.

METHODS AND RESULTS: For these studies, bovine aortic endothelial cells were subjected to physiological levels of equibiaxial cyclic strain (5% strain, 60 cycles/min, 24 hours). In response to strain, both occludin and ZO-1 protein expression increased by 2.3+/-0.1-fold and 2.0+/-0.3-fold, respectively, concomitant with a strain-dependent increase in occludin (but not ZO-1) mRNA levels. These changes were accompanied by reduced occludin tyrosine phosphorylation (75.7+/-8%) and increased ZO-1 serine/threonine phosphorylation (51.7+/-9% and 82.7+/-25%, respectively), modifications that could be completely blocked with tyrosine phosphatase and protein kinase C inhibitors (dephostatin and rottlerin, respectively). In addition, there was a significant strain-dependent increase in endothelial occludin/ZO-1 association (2.0+/-0.1-fold) in parallel with increased localization of both occludin and ZO-1 to the cell-cell border. These events could be completely blocked by dephostatin and rottlerin, and they correlated with a strain-dependent reduction in transendothelial permeability to FITC-dextran.

CONCLUSIONS: Overall, these findings indicate that cyclic strain modulates both the expression and phosphorylation state of occludin and ZO-1 in vascular endothelial cells, with putative consequences for endothelial tight junction assembly and barrier integrity.

Original languageEnglish
Pages (from-to)62-8
Number of pages7
JournalArteriosclerosis, Thrombosis, and Vascular Biology
Volume26
Issue number1
DOIs
Publication statusPublished - Jan 2006

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Occludin
Intercellular Junctions
Tight Junctions
Blood Vessels
Phosphorylation
Endothelial Cells
Hemodynamics
Protein C Inhibitor
Vascular Endothelium
Threonine
Protein Kinase Inhibitors
Phosphoric Monoester Hydrolases
Protein-Tyrosine Kinases
Serine
Protein Kinase C
Endothelium
Tyrosine
Permeability
Pathology
Messenger RNA

Keywords

  • Acetophenones/pharmacology
  • Animals
  • Aorta/cytology
  • Benzopyrans/pharmacology
  • Capillary Permeability/physiology
  • Cattle
  • Dextrans/pharmacokinetics
  • Endothelial Cells/cytology
  • Enzyme Inhibitors/pharmacology
  • Fluorescein-5-isothiocyanate/analogs & derivatives
  • Gene Expression/physiology
  • Hydroquinones/pharmacology
  • In Vitro Techniques
  • Membrane Proteins/genetics
  • Occludin
  • Phosphoproteins/genetics
  • Phosphorylation
  • Protein Kinase C/antagonists & inhibitors
  • Protein Tyrosine Phosphatases/antagonists & inhibitors
  • RNA, Messenger/metabolism
  • Stress, Mechanical
  • Tight Junctions/metabolism
  • Zonula Occludens-1 Protein

Cite this

Collins, Nora T ; Cummins, Philip M ; Colgan, Olga C ; Ferguson, Gail ; Birney, Yvonne A ; Murphy, Ronan P ; Meade, Gerardene ; Cahill, Paul A. / Cyclic strain-mediated regulation of vascular endothelial occludin and ZO-1 : influence on intercellular tight junction assembly and function. In: Arteriosclerosis, Thrombosis, and Vascular Biology. 2006 ; Vol. 26, No. 1. pp. 62-8.
@article{2c4c1542e7084786a72197e6e1711948,
title = "Cyclic strain-mediated regulation of vascular endothelial occludin and ZO-1: influence on intercellular tight junction assembly and function",
abstract = "OBJECTIVE: The vascular endothelium constitutes a highly effective fluid/solute barrier through the regulated apposition of intercellular tight junction complexes. Because endothelium-mediated functions and pathology are driven by hemodynamic forces (cyclic strain and shear stress), we hypothesized a dynamic regulatory link between endothelial tight junction assembly/function and hemodynamic stimuli. We, therefore, examined the effects of cyclic strain on the expression, modification, and function of 2 pivotal endothelial tight junction components, occludin and ZO-1.METHODS AND RESULTS: For these studies, bovine aortic endothelial cells were subjected to physiological levels of equibiaxial cyclic strain (5{\%} strain, 60 cycles/min, 24 hours). In response to strain, both occludin and ZO-1 protein expression increased by 2.3+/-0.1-fold and 2.0+/-0.3-fold, respectively, concomitant with a strain-dependent increase in occludin (but not ZO-1) mRNA levels. These changes were accompanied by reduced occludin tyrosine phosphorylation (75.7+/-8{\%}) and increased ZO-1 serine/threonine phosphorylation (51.7+/-9{\%} and 82.7+/-25{\%}, respectively), modifications that could be completely blocked with tyrosine phosphatase and protein kinase C inhibitors (dephostatin and rottlerin, respectively). In addition, there was a significant strain-dependent increase in endothelial occludin/ZO-1 association (2.0+/-0.1-fold) in parallel with increased localization of both occludin and ZO-1 to the cell-cell border. These events could be completely blocked by dephostatin and rottlerin, and they correlated with a strain-dependent reduction in transendothelial permeability to FITC-dextran.CONCLUSIONS: Overall, these findings indicate that cyclic strain modulates both the expression and phosphorylation state of occludin and ZO-1 in vascular endothelial cells, with putative consequences for endothelial tight junction assembly and barrier integrity.",
keywords = "Acetophenones/pharmacology, Animals, Aorta/cytology, Benzopyrans/pharmacology, Capillary Permeability/physiology, Cattle, Dextrans/pharmacokinetics, Endothelial Cells/cytology, Enzyme Inhibitors/pharmacology, Fluorescein-5-isothiocyanate/analogs & derivatives, Gene Expression/physiology, Hydroquinones/pharmacology, In Vitro Techniques, Membrane Proteins/genetics, Occludin, Phosphoproteins/genetics, Phosphorylation, Protein Kinase C/antagonists & inhibitors, Protein Tyrosine Phosphatases/antagonists & inhibitors, RNA, Messenger/metabolism, Stress, Mechanical, Tight Junctions/metabolism, Zonula Occludens-1 Protein",
author = "Collins, {Nora T} and Cummins, {Philip M} and Colgan, {Olga C} and Gail Ferguson and Birney, {Yvonne A} and Murphy, {Ronan P} and Gerardene Meade and Cahill, {Paul A}",
year = "2006",
month = "1",
doi = "10.1161/01.ATV.0000194097.92824.b3",
language = "English",
volume = "26",
pages = "62--8",
journal = "Arteriosclerosis, Thrombosis, and Vascular Biology",
issn = "1079-5642",
publisher = "Lippincott Williams and Wilkins Ltd.",
number = "1",

}

Cyclic strain-mediated regulation of vascular endothelial occludin and ZO-1 : influence on intercellular tight junction assembly and function. / Collins, Nora T; Cummins, Philip M; Colgan, Olga C; Ferguson, Gail; Birney, Yvonne A; Murphy, Ronan P; Meade, Gerardene; Cahill, Paul A.

In: Arteriosclerosis, Thrombosis, and Vascular Biology, Vol. 26, No. 1, 01.2006, p. 62-8.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Cyclic strain-mediated regulation of vascular endothelial occludin and ZO-1

T2 - influence on intercellular tight junction assembly and function

AU - Collins, Nora T

AU - Cummins, Philip M

AU - Colgan, Olga C

AU - Ferguson, Gail

AU - Birney, Yvonne A

AU - Murphy, Ronan P

AU - Meade, Gerardene

AU - Cahill, Paul A

PY - 2006/1

Y1 - 2006/1

N2 - OBJECTIVE: The vascular endothelium constitutes a highly effective fluid/solute barrier through the regulated apposition of intercellular tight junction complexes. Because endothelium-mediated functions and pathology are driven by hemodynamic forces (cyclic strain and shear stress), we hypothesized a dynamic regulatory link between endothelial tight junction assembly/function and hemodynamic stimuli. We, therefore, examined the effects of cyclic strain on the expression, modification, and function of 2 pivotal endothelial tight junction components, occludin and ZO-1.METHODS AND RESULTS: For these studies, bovine aortic endothelial cells were subjected to physiological levels of equibiaxial cyclic strain (5% strain, 60 cycles/min, 24 hours). In response to strain, both occludin and ZO-1 protein expression increased by 2.3+/-0.1-fold and 2.0+/-0.3-fold, respectively, concomitant with a strain-dependent increase in occludin (but not ZO-1) mRNA levels. These changes were accompanied by reduced occludin tyrosine phosphorylation (75.7+/-8%) and increased ZO-1 serine/threonine phosphorylation (51.7+/-9% and 82.7+/-25%, respectively), modifications that could be completely blocked with tyrosine phosphatase and protein kinase C inhibitors (dephostatin and rottlerin, respectively). In addition, there was a significant strain-dependent increase in endothelial occludin/ZO-1 association (2.0+/-0.1-fold) in parallel with increased localization of both occludin and ZO-1 to the cell-cell border. These events could be completely blocked by dephostatin and rottlerin, and they correlated with a strain-dependent reduction in transendothelial permeability to FITC-dextran.CONCLUSIONS: Overall, these findings indicate that cyclic strain modulates both the expression and phosphorylation state of occludin and ZO-1 in vascular endothelial cells, with putative consequences for endothelial tight junction assembly and barrier integrity.

AB - OBJECTIVE: The vascular endothelium constitutes a highly effective fluid/solute barrier through the regulated apposition of intercellular tight junction complexes. Because endothelium-mediated functions and pathology are driven by hemodynamic forces (cyclic strain and shear stress), we hypothesized a dynamic regulatory link between endothelial tight junction assembly/function and hemodynamic stimuli. We, therefore, examined the effects of cyclic strain on the expression, modification, and function of 2 pivotal endothelial tight junction components, occludin and ZO-1.METHODS AND RESULTS: For these studies, bovine aortic endothelial cells were subjected to physiological levels of equibiaxial cyclic strain (5% strain, 60 cycles/min, 24 hours). In response to strain, both occludin and ZO-1 protein expression increased by 2.3+/-0.1-fold and 2.0+/-0.3-fold, respectively, concomitant with a strain-dependent increase in occludin (but not ZO-1) mRNA levels. These changes were accompanied by reduced occludin tyrosine phosphorylation (75.7+/-8%) and increased ZO-1 serine/threonine phosphorylation (51.7+/-9% and 82.7+/-25%, respectively), modifications that could be completely blocked with tyrosine phosphatase and protein kinase C inhibitors (dephostatin and rottlerin, respectively). In addition, there was a significant strain-dependent increase in endothelial occludin/ZO-1 association (2.0+/-0.1-fold) in parallel with increased localization of both occludin and ZO-1 to the cell-cell border. These events could be completely blocked by dephostatin and rottlerin, and they correlated with a strain-dependent reduction in transendothelial permeability to FITC-dextran.CONCLUSIONS: Overall, these findings indicate that cyclic strain modulates both the expression and phosphorylation state of occludin and ZO-1 in vascular endothelial cells, with putative consequences for endothelial tight junction assembly and barrier integrity.

KW - Acetophenones/pharmacology

KW - Animals

KW - Aorta/cytology

KW - Benzopyrans/pharmacology

KW - Capillary Permeability/physiology

KW - Cattle

KW - Dextrans/pharmacokinetics

KW - Endothelial Cells/cytology

KW - Enzyme Inhibitors/pharmacology

KW - Fluorescein-5-isothiocyanate/analogs & derivatives

KW - Gene Expression/physiology

KW - Hydroquinones/pharmacology

KW - In Vitro Techniques

KW - Membrane Proteins/genetics

KW - Occludin

KW - Phosphoproteins/genetics

KW - Phosphorylation

KW - Protein Kinase C/antagonists & inhibitors

KW - Protein Tyrosine Phosphatases/antagonists & inhibitors

KW - RNA, Messenger/metabolism

KW - Stress, Mechanical

KW - Tight Junctions/metabolism

KW - Zonula Occludens-1 Protein

U2 - 10.1161/01.ATV.0000194097.92824.b3

DO - 10.1161/01.ATV.0000194097.92824.b3

M3 - Article

C2 - 16269664

VL - 26

SP - 62

EP - 68

JO - Arteriosclerosis, Thrombosis, and Vascular Biology

JF - Arteriosclerosis, Thrombosis, and Vascular Biology

SN - 1079-5642

IS - 1

ER -