Rho flares repair local tight junction leaks

Rachel E Stephenson, Tomohito Higashi, Ivan S. Erofeev, Torey R. Arnold, Marcin Leda, Andrew B. Goryachev, Ann L. Miller

Research output: Contribution to journalArticlepeer-review


Tight junctions contribute to epithelial barrier function by selectively regulating the quantity and type of molecules that cross the paracellular barrier. Experimental approaches to evaluate the effectiveness of tight junctions are typically global, tissue-scale measures. Here, we introduce Zincbased Ultrasensitive Microscopic Barrier Assay (ZnUMBA), which we used in Xenopus laevis embryos to visualize short-lived, local breaches in epithelial barrier function. These breaches, or leaks, occur as cell boundaries elongate, correspond to visible breaks in the tight junction, and are followed by transient localized Rho activation, or Rho flares. We discovered that Rho flares restore barrier function by driving concentration of tight junction proteins through actin polymerization and ROCK-mediated localized contraction of the cell boundary. We conclude that Rho flares constitute a damage control mechanism that reinstates barrier function when tight junctions become locally compromised due to normallyoccurring changes in cell shape and tissue tension.
Original languageEnglish
Pages (from-to)445-459
Number of pages20
JournalDevelopmental Cell
Issue number4
Early online date14 Feb 2019
Publication statusPublished - 25 Feb 2019


  • tight junction
  • barrier assay
  • leak pathway
  • Rho GTPase
  • actin
  • myosin II
  • junction contraction
  • epithelium
  • Xenopus


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