Cdc42 controls vascular network assembly through protein kinase Cι during embryonic vasculogenesis.
Cdc42 controls vascular network assembly through protein kinase Cι during embryonic vasculogenesis.
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Cdc42 在胚胎血管发生过程中通过蛋白激酶 Cδ 控制血管网络组装。
DOI：
10.1161/atvbaha.111.230144
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发表时间：
2011
期刊：
影响因子：
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通讯作者：
Li,Shaohua
中科院分区：
文献类型：
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作者：
Qi,Yanmei;Liu,Jie;Wu,Xunwei;Brakebusch,Cord;Leitges,Michael;Han,Yaling;Corbett,SiobhanA;Lowry,StephenF;Graham,AlanM;Li,Shaohua
目的研究Cdc42在胚胎血管发生中的作用及其机制。方法和结果通过使用转基因小鼠胚胎干（ES）细胞，我们证明了Rho GTPase Cdc42的去除可以阻断胚状体（EB）血管形成过程中的血管网络组装，而不影响内皮细胞谱系的分化。突变体EBs中Cdc42的重表达挽救了突变体的表型，确立了Cdc42在血管发生中的重要作用。嵌合分析显示，血管表型是由内皮细胞而不是周围细胞的Cdc42失活引起的。从cdc42缺失的EBs中分离的内皮细胞在定向迁移和网络组装方面存在缺陷。此外，在cdc42缺失的内皮细胞中，非典型蛋白激酶Cι (PKCι)的激活被取消，PKCι消融表现了cdc42缺失的EBs的血管异常。此外，糖原合成酶激酶-3β (GSK-3β) Ser9位点的抑制性磷酸化依赖于Cdc42和PKCι，在Cdc42缺失的EBs中，激酶死亡的GSK-3β的表达促进了无分支的线性内皮节段的形成。这些结果表明PKCι和GSK-3β是血管形态发生过程中Cdc42的下游效应物。结论在胚胎血管发生过程中，cdc42通过激活pkc1来控制血管网络的组装，而不是内皮谱系的分化。
ObjectiveThe goal of this study was to determine the role of Cdc42 in embryonic vasculogenesis and the underlying mechanisms.Methods and ResultsBy using genetically modified mouse embryonic stem (ES) cells, we demonstrate that ablation of the Rho GTPase Cdc42 blocks vascular network assembly during embryoid body (EB) vasculogenesis without affecting endothelial lineage differentiation. Reexpression of Cdc42 in mutant EBs rescues the mutant phenotype, establishing an essential role for Cdc42 in vasculogenesis. Chimeric analysis revealed that the vascular phenotype is caused by inactivation of Cdc42 in endothelial cells rather than surrounding cells. Endothelial cells isolated from Cdc42-null EBs are defective in directional migration and network assembly. In addition, activation of atypical protein kinase Cι (PKCι) is abolished in Cdc42-null endothelial cells, and PKCι ablation phenocopies the vascular abnormalities of the Cdc42-null EBs. Moreover, the inhibitory phosphorylation of glycogen synthase kinase-3β (GSK-3β) at Ser9 depends on Cdc42 and PKCι, and expression of kinase-dead GSK-3β in Cdc42-null EBs promotes the formation of linear endothelial segments without branches. These results suggest that PKCι and GSK-3β are downstream effectors of Cdc42 during vascular morphogenesis.ConclusionCdc42 controls vascular network assembly but not endothelial lineage differentiation by activating PKCι during embryonic vasculogenesis.