title: An Adaptor Molecule Afadin Regulates Lymphangiogenesis by Modulating RhoA Activity in the Developing Mouse Embryo.
abstract: Afadin is an intracellular binding partner of nectins, cell-cell adhesion molecules, and plays important roles in the formation of cell-cell junctions. Afadin-knockout mice show early embryonic lethality, therefore little is known about the function of afadin during organ development. In this study, we generated mice lacking afadin expression in endothelial cells, and found that the majority of these mice were embryonically lethal as a result of severe subcutaneous edema. Defects in the lymphatic vessels of the skin were observed, although the morphology in the blood vessels was almost normal. Severe disruption of VE-cadherin-mediated cell-cell junctions occurred only in lymphatic endothelial cells, but not in blood endothelial cells. Knockout of afadin did not affect the differentiation and proliferation of lymphatic endothelial cells. Using in vitro assays with blood and lymphatic microvascular endothelial cells (BMVECs and LMVECs, respectively), knockdown of afadin caused elongated cell shapes and disruption of cell-cell junctions among LMVECs, but not BMVECs. In afadin-knockdown LMVECs, enhanced F-actin bundles at the cell periphery and reduced VE-cadherin immunostaining were found, and activation of RhoA was strongly increased compared with that in afadin-knockdown BMVECs. Conversely, inhibition of RhoA activation in afadin-knockdown LMVECs restored the cell morphology. These results indicate that afadin has different effects on blood and lymphatic endothelial cells by controlling the levels of RhoA activation, which may critically regulate the lymphangiogenesis of mouse embryos.
authors.affiliation: Department of Molecular Biology, Osaka Medical Center for Cancer and Cardiovascular Disease, Osaka, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Majima
authors.forename: Keisuke
authors.initials: K
authors.lastname: Takeuchi
authors.forename: Keigo
authors.initials: K
authors.lastname: Sano
authors.forename: Masanori
authors.initials: M
authors.lastname: Hirashima
authors.forename: Dimitar P
authors.initials: DP
authors.lastname: Zankov
authors.forename: Miki
authors.initials: M
authors.lastname: Tanaka-Okamoto
authors.forename: Hiroyoshi
authors.initials: H
authors.lastname: Ishizaki
authors.forename: Jun
authors.initials: J
authors.lastname: Miyoshi
authors.forename: Hisakazu
authors.initials: H
authors.lastname: Ogita
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology* / Q000378:metabolism; D047108:Embryonic Development; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D007365:Intercellular Junctions / Q000502:physiology; D042583:Lymphangiogenesis / Q000502:physiology*; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008840:Microfilament Proteins / Q000502:physiology*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
chemical_list: D008840:Microfilament Proteins; C108835:afadin; D020742:rhoA GTP-Binding Protein