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Isolation of Primary Murine Brain Microvascular Endothelial Cells
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Published on: November 14, 2014

Bves modulates tight junction associated signaling.

Patricia K Russ1, Christopher J Pino, Christopher S Williams

  • 1Department of Biomedical Engineering, Vanderbilt University, Nashville, Tennessee, United States of America. patricia.k.russ@vanderbilt.edu

Plos One
|February 2, 2011
PubMed
Summary
This summary is machine-generated.

Blood vessel epicardial substance (Bves) regulates epithelial tight junctions (TJs). Truncated Bves disrupts TJs, increasing RhoA activity and ZONAB/DbpA transcription, while full-length Bves enhances TJ function.

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Area of Science:

  • Cell Biology
  • Epithelial Biology
  • Molecular Biology

Background:

  • Blood vessel epicardial substance (Bves) is a transmembrane adhesion protein crucial for epithelial tight junction (TJ) formation.
  • TJs are vital for epithelial barrier function and cellular signaling, influencing pathways like RhoA and ZONAB/DbpA.
  • The precise role of Bves in modulating these signaling pathways via TJ regulation remains to be fully elucidated.

Purpose of the Study:

  • To investigate the hypothesis that Bves modulates RhoA activation and ZONAB/DbpA activity through its regulation of TJ formation.
  • To examine the effects of full-length and truncated Bves on TJ integrity and associated signaling pathways in human corneal epithelial (HCE) cells.

Main Methods:

  • Stable transfection of HCE cells with full-length (w-Bves) or C-terminus truncated (t-Bves) Flag-tagged chicken Bves.
  • Assessment of Bves interaction with endogenous Bves and ZO-1.
  • Measurement of trans-epithelial electrical resistance (TEER) to evaluate TJ function.
  • Immunolocalization of TJ proteins, ZONAB/DbpA, and GEF-H1.
  • FRET-based RhoA activity assays and luciferase reporter assays for ZONAB/DbpA transcriptional activity.

Main Results:

  • Both w-Bves and t-Bves interacted with endogenous Bves, but t-Bves disrupted membrane localization and ZO-1 interaction.
  • w-Bves cells showed increased TEER and proper TJ protein localization, while t-Bves cells exhibited decreased TEER and loss of TJ protein localization.
  • t-Bves expression led to decreased membrane localization of ZONAB/DbpA and GEF-H1.
  • t-Bves cells displayed a significant 30% increase in RhoA activity and elevated ZONAB/DbpA transcriptional activity compared to controls.

Conclusions:

  • Bves expression and proper localization are critical for maintaining epithelial TJ integrity and function.
  • Truncated Bves disrupts TJs, leading to aberrant RhoA activation and enhanced ZONAB/DbpA transcriptional activity.
  • These findings highlight Bves as a key regulator of epithelial barrier function and cellular signaling.