title: Stabilization of brain microvascular endothelial barrier function by shear stress involves VE-cadherin signaling leading to modulation of pTyr-occludin levels.
abstract: Blood-brain barrier (BBB) regulation involves the coordinated interaction of intercellular adherens and tight junctions in response to stimuli. One such stimulus, shear stress, has been shown to upregulate brain microvascular endothelial cell (BMvEC) barrier function, although our knowledge of the signaling mechanisms involved is limited. In this article, we examined the hypothesis that VE-cadherin can transmit shear signals to tight junction occludin with consequences for pTyr-occludin and barrier function. In initial studies, chronic shear enhanced membrane localization of ZO-1 and claudin-5, decreased pTyr-occludin (in part via a dephostatin-sensitive mechanism), and reduced BMvEC permeability, with flow reduction in pre-sheared BMvECs having converse effects. In further studies, VE-cadherin inhibition (VE-cad ΔEXD) blocked shear-induced Rac1 activation, pTyr-occludin reduction, and barrier upregulation, consistent with an upstream role for VE-cadherin in transmitting shear signals to tight junctions through Rac1. As VE-cadherin is known to mediate Rac1 activation via Tiam1 recruitment, we subsequently confirmed that Tiam1 inhibition (Tiam1-C580) could elicit effects similar to VE-cad ΔEXD. Finally, the observed attenuation of shear-induced changes in pTyr-occludin level and barrier phenotype following Rac1 inhibition (NSC23766, T17N) establishes a downstream role for Rac1 in this pathway. In summary, we describe for the first time in BMvECs a role for VE-cadherin in the transmission of physiological shear signals to tight junction occludin through engagement of Tiam1/Rac1 leading to barrier stabilization. A downstream role is also strongly indicated for a protein tyrosine phosphatase in pTyr-occludin modulation. Importantly, these findings suggest an important route of inter-junctional signaling cross-talk during BBB response to flow.
authors.affiliation: School of Biotechnology, Dublin City University, Glasnevin, Dublin 9, Ireland.
authors.forename: Tony G
authors.initials: TG
authors.lastname: Walsh
authors.forename: Ronan P
authors.initials: RP
authors.lastname: Murphy
authors.forename: Paul
authors.initials: P
authors.lastname: Fitzpatrick
authors.forename: Keith D
authors.initials: KD
authors.lastname: Rochfort
authors.forename: Anthony F
authors.initials: AF
authors.lastname: Guinan
authors.forename: Andrew
authors.initials: A
authors.lastname: Murphy
authors.forename: Philip M
authors.initials: PM
authors.lastname: Cummins
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D001812:Blood-Brain Barrier / Q000502:physiology*; D015820:Cadherins / Q000378:metabolism*; D002199:Capillary Permeability / Q000502:physiology; D002417:Cattle; D062507:Claudin-5; D004730:Endothelium, Vascular / Q000378:metabolism*; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D055806:Microvessels / Q000378:metabolism; D062793:Occludin; D010750:Phosphoproteins / Q000378:metabolism; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism; D033081:Shear Strength; D015398:Signal Transduction; D013314:Stress, Mechanical; D000075684:T-Lymphoma Invasion and Metastasis-inducing Protein 1; D019108:Tight Junctions / Q000502:physiology; D062826:Zonula Occludens-1 Protein; D020830:rac1 GTP-Binding Protein / Q000378:metabolism; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D062507:Claudin-5; D020662:Guanine Nucleotide Exchange Factors; D008565:Membrane Proteins; D062793:Occludin; D010750:Phosphoproteins; D017027:Protein Tyrosine Phosphatases; D000075684:T-Lymphoma Invasion and Metastasis-inducing Protein 1; D062826:Zonula Occludens-1 Protein; D020830:rac1 GTP-Binding Protein; D000099152:Cadherin 5; C107466:CLDN5 protein, human; C568985:OCLN protein, human; C087334:TIAM1 protein, human; C569042:TJP1 protein, human