Division of Developmental Biology, National Institute of Child Health and Human Development, National Institutes of Health, 6 Center Dr. Bethesda, Bethesda, MD, 20892, USA.
Department of Cell and Molecular Biology, University of Pennsylvania, 440 Curie Blvd, Philadelphia, PA, 19104, USA.
Angiogenesis. 2022 Aug;25(3):411-434. doi: 10.1007/s10456-022-09834-9. Epub 2022 Mar 23.
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.
小分子单体 GTPase RHOA 通过激活细胞信号转导级联反应的效应物,包括 Rho 相关蛋白激酶 ROCK1/2,充当信号转导级联反应的主要调节剂。先前的体外细胞培养研究表明,RHOA 可以调节血管内皮细胞 (EC) 生物学的许多关键方面,包括焦点黏附、应激纤维形成和血管生成。然而,RHOA 在血管发育和稳态中的具体体内作用仍未得到很好的理解。在这项研究中,我们研究了 RHOA 在调节斑马鱼血管发育和完整性中的体内功能。我们使用斑马鱼 RHOA 同源物 (rhoaa) 突变体、在 EC 中表达野生型、显性负性或组成性激活形式的 rhoaa 的转基因胚胎、RHOA 和 ROCK1/2 的药理学抑制剂以及 Rock1 和 Rock2a/b dgRNP 注射的斑马鱼胚胎,研究 RHOA 在血管内皮细胞中的功能获得和功能丧失的体内后果。我们的研究结果证明了 RHOA 在血管完整性、发育性血管生成和体内血管形态发生中的作用,表明 RHOA 活性过多或过少都会导致血管功能障碍。