Cdc42 Controls Vascular Network Assembly Through Protein Kinase C{iota} During Embryonic Vasculogenesis

TY - JOUR

T1 - Cdc42 Controls Vascular Network Assembly Through Protein Kinase C{iota} During Embryonic Vasculogenesis

AU - Qi, Yanmei

AU - Liu, Jie

AU - Wu, Xunwei

AU - Brakebusch, Cord

AU - Leitges, Michael

AU - Han, Yaling

AU - Corbett, Siobhan A

AU - Lowry, Stephen F

AU - Graham, Alan M

AU - Li, Shaohua

PY - 2011/8/1

Y1 - 2011/8/1

N2 - Objective-: The goal of this study was to determine the role of Cdc42 in embryonic vasculogenesis and the underlying mechanisms. Methods and results-: By 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. Conclusion-: Cdc42 controls vascular network assembly but not endothelial lineage differentiation by activating PKCι during embryonic vasculogenesis.

AB - Objective-: The goal of this study was to determine the role of Cdc42 in embryonic vasculogenesis and the underlying mechanisms. Methods and results-: By 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. Conclusion-: Cdc42 controls vascular network assembly but not endothelial lineage differentiation by activating PKCι during embryonic vasculogenesis.

U2 - 10.1161/ATVBAHA.111.230144

DO - 10.1161/ATVBAHA.111.230144

M3 - Journal article

C2 - 21659643

SN - 1079-5642

VL - 31

SP - 1861

EP - 1870

JO - Arteriosclerosis, Thrombosis, and Vascular Biology

JF - Arteriosclerosis, Thrombosis, and Vascular Biology

IS - 8

ER -