In vivo dissection of Rhoa function in vascular development using zebrafish

Laura M. Pillay, Joseph J. Yano, Andrew E. Davis, Matthew G. Butler, Megan O. Ezeude, Jong S. Park, Keith A. Barnes, Vanessa L. Reyes, Daniel Castranova, Aniket V. Gore, Matthew R. Swift, James R. Iben, Madeleine I. Kenton, Amber N. Stratman, Brant M. Weinstein

Research output: Contribution to journalArticlepeer-review

6 Scopus citations


The small monomeric GTPase RHOA acts as a master regulator of signal transduction cascades by activating effectors of cellular signaling, including the Rho-associated protein kinases ROCK1/2. Previous in vitro cell culture studies suggest that RHOA can regulate many critical aspects of vascular endothelial cell (EC) biology, including focal adhesion, stress fiber formation, and angiogenesis. However, the specific in vivo roles of RHOA during vascular development and homeostasis are still not well understood. In this study, we examine the in vivo functions of RHOA in regulating vascular development and integrity in zebrafish. We use zebrafish RHOA-ortholog (rhoaa) mutants, transgenic embryos expressing wild type, dominant negative, or constitutively active forms of rhoaa in ECs, pharmacological inhibitors of RHOA and ROCK1/2, and Rock1 and Rock2a/b dgRNP-injected zebrafish embryos to study the in vivo consequences of RHOA gain- and loss-of-function in the vascular endothelium. Our findings document roles for RHOA in vascular integrity, developmental angiogenesis, and vascular morphogenesis in vivo, showing that either too much or too little RHOA activity leads to vascular dysfunction.

Original languageEnglish
Pages (from-to)411-434
Number of pages24
Issue number3
StatePublished - Aug 2022


  • Angiogenesis
  • Endothelial cell
  • Hemorrhage
  • Rhoa
  • Rock
  • Zebrafish


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