The Cdc42 and Rac1 GTPases are required for capillary lumen formation in three-dimensional extracellular matrices.
The Cdc42 and Rac1 GTPases are required for capillary lumen formation in three-dimensional extracellular matrices.
复制标题
DOI：
--
复制
发表时间：
2002-03
影响因子：
4
通讯作者：
K. Bayless;G. Davis
中科院分区：
文献类型：
--
作者：
K. Bayless;G. Davis
在这里，我们显示了在三维细胞外基质的内皮细胞（EC）形态发生的Cdc 42和Rac 1 GTP酶的要求。Cdc 42和Rac 1特异性调节EC细胞内空泡和管腔形成的胶原蛋白和纤维蛋白基质。艰难梭菌毒素B（阻断所有三种Rho GTP酶）完全抑制EC形成空泡和管腔的能力，而Rho的选择性抑制剂C3转移酶则没有。无论是显性阴性（N17）或组成型活性（V12）Cdc 42使用重组腺病毒的表达显着抑制EC空泡和管腔形成的胶原蛋白和纤维蛋白基质。空泡和管腔的形成开始于表达显性阴性（N17）Rac 1的EC，但后来塌陷，表明Rac 1在血管发育后期的作用。使用共聚焦显微镜分析的文化揭示了绿色荧光蛋白-V12 Rac 1，-Rac 1野生型和-Cdc 42野生型嵌合蛋白质的细胞内空泡膜在管腔形成过程中的目标。此外，表达的verprolin-cofilin-酸性结构域的N-WASP，下游Cdc 42效应，在EC中完全干扰空泡和管腔的形成。这些结果共同揭示了Cdc 42和Rac 1在三维细胞外基质中EC空泡和管腔形成过程中的新作用。
Here we show a requirement for the Cdc42 and Rac1 GTPases in endothelial cell (EC) morphogenesis in three-dimensional extracellular matrices. Cdc42 and Rac1 specifically regulate EC intracellular vacuole and lumen formation in both collagen and fibrin matrices. Clostridium difficile toxin B (which blocks all three Rho GTPases) completely inhibited the ability of ECs to form both vacuoles and lumens, whereas C3 transferase, a selective inhibitor of Rho, did not. Expression of either dominant-negative (N17) or constitutively active (V12) Cdc42 using recombinant adenoviruses dramatically inhibited EC vacuole and lumen formation in both collagen and fibrin matrices. Both vacuole and lumen formation initiated in ECs expressing dominant-negative (N17) Rac1 but later collapsed, indicating a role for Rac1 during later stages of vessel development. Analysis of cultures using confocal microscopy revealed green fluorescent protein-V12Rac1, -Rac1 wild-type and -Cdc42 wild-type chimeric proteins targeted to intracellular vacuole membranes during the lumen formation process. Also, expression of the verprolin-cofilin-acidic domain of N-WASP, a downstream Cdc42 effector, in ECs completely interfered with vacuole and lumen formation. These results collectively reveal a novel role for Cdc42 and Rac1 in the process of EC vacuole and lumen formation in three-dimensional extracellular matrices.