Regulation of bovine brain microvascular endothelial tight junction assembly and barrier function by laminar shear stress

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

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

Blood-brain barrier (BBB) controls paracellular solute diffusion into the brain microenvironment and is maintained primarily by tight junctions between adjacent microvascular endothelial cells. Studies implicate blood flow-associated shear stress as a pathophysiological mediator of BBB function, although detailed biochemical data are scarce. We hypothesize that shear stress upregulates BBB function via direct modulation of expression and properties of pivotal tight-junction proteins occludin and zonula occludens-1 (ZO-1). Bovine brain microvascular endothelial cells (BBMvECs) were exposed to either steady or pulsatile shear stress (10 and 14 dyn/cm(2), respectively) for 24 h. Sheared BBMvECs were monitored for occludin-ZO-1 expression, association, and subcellular localization, and transendothelial permeability of BBMvECs to FITC-dextran and (14)[C]sucrose was assessed. Actin reorganization and BBMvEC realignment were observed following steady shear stress for 24 h. Substantial increases in occludin mRNA and protein expression (2.73 +/- 0.26- and 1.83 +/- 0.03-fold) and in occludin-ZO-1 association (2.12 +/- 0.15-fold) were also observed. Steady shear stress also induced clear relocalization of both proteins to the cell-cell border in parallel with reduced transendothelial permeability to FITC-dextran (but not sucrose). Following pulsatile shear stress, increased protein levels for both occludin and ZO-1 (2.15 +/- 0.02- and 1.67 +/- 0.21-fold) and increased occludin-ZO-1 association (2.91 +/- 0.14-fold) were observed in parallel with a reduction in transendothelial permeability to (14)[C]sucrose. Shear stress upregulates BBMvEC barrier function at the molecular level via modulation of expression, association, and localization of occludin and ZO-1. The pulsatile shear model appeared to give the most profound biochemical responses.

Original languageEnglish
Pages (from-to)H3190-7
JournalAmerican Journal of Physiology - Heart and Circulatory Physiology
Volume292
Issue number6
DOIs
Publication statusPublished - Jun 2007

Fingerprint

Occludin
Tight Junctions
Endothelial Cells
Brain
Blood-Brain Barrier
Sucrose
Permeability
Up-Regulation
Tight Junction Proteins
Proteins
Actins
Messenger RNA

Keywords

  • Actins/metabolism
  • Animals
  • Blood-Brain Barrier/cytology
  • Brain/blood supply
  • Capillary Permeability
  • Cattle
  • Cell Shape
  • Cells, Cultured
  • Dextrans/metabolism
  • Endothelial Cells/metabolism
  • Fluorescein-5-isothiocyanate/analogs & derivatives
  • Membrane Proteins/genetics
  • Microcirculation/cytology
  • Occludin
  • Phosphoproteins/genetics
  • Pulsatile Flow
  • RNA, Messenger/metabolism
  • Stress, Mechanical
  • Sucrose/metabolism
  • Tight Junctions/metabolism
  • Zonula Occludens-1 Protein

Cite this

Colgan, Olga C ; Ferguson, Gail ; Collins, Nora T ; Murphy, Ronan P ; Meade, Gerardeane ; Cahill, Paul A ; Cummins, Philip M. / Regulation of bovine brain microvascular endothelial tight junction assembly and barrier function by laminar shear stress. In: American Journal of Physiology - Heart and Circulatory Physiology. 2007 ; Vol. 292, No. 6. pp. H3190-7.
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abstract = "Blood-brain barrier (BBB) controls paracellular solute diffusion into the brain microenvironment and is maintained primarily by tight junctions between adjacent microvascular endothelial cells. Studies implicate blood flow-associated shear stress as a pathophysiological mediator of BBB function, although detailed biochemical data are scarce. We hypothesize that shear stress upregulates BBB function via direct modulation of expression and properties of pivotal tight-junction proteins occludin and zonula occludens-1 (ZO-1). Bovine brain microvascular endothelial cells (BBMvECs) were exposed to either steady or pulsatile shear stress (10 and 14 dyn/cm(2), respectively) for 24 h. Sheared BBMvECs were monitored for occludin-ZO-1 expression, association, and subcellular localization, and transendothelial permeability of BBMvECs to FITC-dextran and (14)[C]sucrose was assessed. Actin reorganization and BBMvEC realignment were observed following steady shear stress for 24 h. Substantial increases in occludin mRNA and protein expression (2.73 +/- 0.26- and 1.83 +/- 0.03-fold) and in occludin-ZO-1 association (2.12 +/- 0.15-fold) were also observed. Steady shear stress also induced clear relocalization of both proteins to the cell-cell border in parallel with reduced transendothelial permeability to FITC-dextran (but not sucrose). Following pulsatile shear stress, increased protein levels for both occludin and ZO-1 (2.15 +/- 0.02- and 1.67 +/- 0.21-fold) and increased occludin-ZO-1 association (2.91 +/- 0.14-fold) were observed in parallel with a reduction in transendothelial permeability to (14)[C]sucrose. Shear stress upregulates BBMvEC barrier function at the molecular level via modulation of expression, association, and localization of occludin and ZO-1. The pulsatile shear model appeared to give the most profound biochemical responses.",
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Regulation of bovine brain microvascular endothelial tight junction assembly and barrier function by laminar shear stress. / Colgan, Olga C; Ferguson, Gail; Collins, Nora T; Murphy, Ronan P; Meade, Gerardeane; Cahill, Paul A; Cummins, Philip M.

In: American Journal of Physiology - Heart and Circulatory Physiology, Vol. 292, No. 6, 06.2007, p. H3190-7.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Regulation of bovine brain microvascular endothelial tight junction assembly and barrier function by laminar shear stress

AU - Colgan, Olga C

AU - Ferguson, Gail

AU - Collins, Nora T

AU - Murphy, Ronan P

AU - Meade, Gerardeane

AU - Cahill, Paul A

AU - Cummins, Philip M

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N2 - Blood-brain barrier (BBB) controls paracellular solute diffusion into the brain microenvironment and is maintained primarily by tight junctions between adjacent microvascular endothelial cells. Studies implicate blood flow-associated shear stress as a pathophysiological mediator of BBB function, although detailed biochemical data are scarce. We hypothesize that shear stress upregulates BBB function via direct modulation of expression and properties of pivotal tight-junction proteins occludin and zonula occludens-1 (ZO-1). Bovine brain microvascular endothelial cells (BBMvECs) were exposed to either steady or pulsatile shear stress (10 and 14 dyn/cm(2), respectively) for 24 h. Sheared BBMvECs were monitored for occludin-ZO-1 expression, association, and subcellular localization, and transendothelial permeability of BBMvECs to FITC-dextran and (14)[C]sucrose was assessed. Actin reorganization and BBMvEC realignment were observed following steady shear stress for 24 h. Substantial increases in occludin mRNA and protein expression (2.73 +/- 0.26- and 1.83 +/- 0.03-fold) and in occludin-ZO-1 association (2.12 +/- 0.15-fold) were also observed. Steady shear stress also induced clear relocalization of both proteins to the cell-cell border in parallel with reduced transendothelial permeability to FITC-dextran (but not sucrose). Following pulsatile shear stress, increased protein levels for both occludin and ZO-1 (2.15 +/- 0.02- and 1.67 +/- 0.21-fold) and increased occludin-ZO-1 association (2.91 +/- 0.14-fold) were observed in parallel with a reduction in transendothelial permeability to (14)[C]sucrose. Shear stress upregulates BBMvEC barrier function at the molecular level via modulation of expression, association, and localization of occludin and ZO-1. The pulsatile shear model appeared to give the most profound biochemical responses.

AB - Blood-brain barrier (BBB) controls paracellular solute diffusion into the brain microenvironment and is maintained primarily by tight junctions between adjacent microvascular endothelial cells. Studies implicate blood flow-associated shear stress as a pathophysiological mediator of BBB function, although detailed biochemical data are scarce. We hypothesize that shear stress upregulates BBB function via direct modulation of expression and properties of pivotal tight-junction proteins occludin and zonula occludens-1 (ZO-1). Bovine brain microvascular endothelial cells (BBMvECs) were exposed to either steady or pulsatile shear stress (10 and 14 dyn/cm(2), respectively) for 24 h. Sheared BBMvECs were monitored for occludin-ZO-1 expression, association, and subcellular localization, and transendothelial permeability of BBMvECs to FITC-dextran and (14)[C]sucrose was assessed. Actin reorganization and BBMvEC realignment were observed following steady shear stress for 24 h. Substantial increases in occludin mRNA and protein expression (2.73 +/- 0.26- and 1.83 +/- 0.03-fold) and in occludin-ZO-1 association (2.12 +/- 0.15-fold) were also observed. Steady shear stress also induced clear relocalization of both proteins to the cell-cell border in parallel with reduced transendothelial permeability to FITC-dextran (but not sucrose). Following pulsatile shear stress, increased protein levels for both occludin and ZO-1 (2.15 +/- 0.02- and 1.67 +/- 0.21-fold) and increased occludin-ZO-1 association (2.91 +/- 0.14-fold) were observed in parallel with a reduction in transendothelial permeability to (14)[C]sucrose. Shear stress upregulates BBMvEC barrier function at the molecular level via modulation of expression, association, and localization of occludin and ZO-1. The pulsatile shear model appeared to give the most profound biochemical responses.

KW - Actins/metabolism

KW - Animals

KW - Blood-Brain Barrier/cytology

KW - Brain/blood supply

KW - Capillary Permeability

KW - Cattle

KW - Cell Shape

KW - Cells, Cultured

KW - Dextrans/metabolism

KW - Endothelial Cells/metabolism

KW - Fluorescein-5-isothiocyanate/analogs & derivatives

KW - Membrane Proteins/genetics

KW - Microcirculation/cytology

KW - Occludin

KW - Phosphoproteins/genetics

KW - Pulsatile Flow

KW - RNA, Messenger/metabolism

KW - Stress, Mechanical

KW - Sucrose/metabolism

KW - Tight Junctions/metabolism

KW - Zonula Occludens-1 Protein

U2 - 10.1152/ajpheart.01177.2006

DO - 10.1152/ajpheart.01177.2006

M3 - Article

C2 - 17308001

VL - 292

SP - H3190-7

JO - American Journal of Physiology - Heart and Circulatory Physiology

JF - American Journal of Physiology - Heart and Circulatory Physiology

SN - 0363-6135

IS - 6

ER -