title: Dynamics of planar cell polarity protein Vangl2 in the mouse oviduct epithelium.
abstract: The planar cell polarity (PCP) pathway regulates morphogenesis in various organs. The polarized localization is a key feature of core PCP factors for orchestrating cell polarity in an epithelial sheet. Several studies using Drosophila melanogaster have investigated the mechanism of the polarized localization. However, to what extent these mechanisms are conserved and how the polarization of core PCP factors is maintained in mature vertebrates are still open questions. Here, we addressed these questions by analyzing the dynamics of Vangl2, a member of core PCP factors, in the mouse oviduct epithelium. Multiple core PCP factors including Vangl2 were expressed in the mouse oviduct in postnatal stages. Vangl1, Vangl2 and Frizzled6 had polarized localization in the oviduct epithelium. Exogenously introduced expression of green fluorescent protein (GFP)-tagged core PCP factors by electroporation revealed that Vangl1, Vangl2 and Prickle2 are localized on the ovarian side of the cell periphery in the oviduct. To visualize the Vangl2 dynamics, we generated the R26-Vangl2-EGFP transgenic mice. In these mice, Vangl2-EGFP was ubiquitously expressed and showed polarized localization in multiple organs including the oviduct, the trachea, the lateral ventricle and the uterus. Fluorescence recovery after photobleaching (FRAP) analysis in the mature oviduct revealed that Vangl2 in the enriched subdomain of cell periphery (cellular edge) was more stable than Vangl2 in the less-enriched cellular edge. Furthermore, when a subregion of a Vangl2-enriched cellular edge was bleached, the Vangl2-enriched subregion neighboring the bleached region in the same cellular edge tended to decrease more intensities than the neighboring sub-region in the next Vangl2-enriched cellular edge. Finally, the polarization of Vangl2 was observed in nocodazole treated mouse viduct, suggesting the maintenance of Vangl2 asymmetry is independent of microtubule formation. Taken together, we revealed the characteristics of Vangl2 dynamics in the oviduct epithelium, and found that Vangl2 forms stable complex at the enriched cellular edge and forms compartments. Our data collectively suggest that the mechanism for maintenance of Vangl2 asymmetry in mature mouse oviduct is different from the microtubule dependent polarized transport model, which has been proposed for the reinforcement of the asymmetry of two core PCP proteins, Flamingo and Dishevelled, in the developing fly.
authors.affiliation: Division of Embryology, National Institute for Basic Biology, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, Japan; Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan. Electronic address: thseki@gmail.com.
authors.forename: Dongbo
authors.initials: D
authors.lastname: Shi
authors.affiliation: Division of Embryology, National Institute for Basic Biology, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, Japan; Department of Basic Biology, SOKENDAI (The Graduate University for Advanced Studies), Japan.
authors.forename: Fumiko
authors.initials: F
authors.lastname: Usami
authors.affiliation: Division of Embryology, National Institute for Basic Biology, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, Japan.
authors.forename: Kouji
authors.initials: K
authors.lastname: Komatsu
authors.affiliation: Division of Embryology, National Institute for Basic Biology, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, Japan.
authors.forename: Sanae
authors.initials: S
authors.lastname: Oka
authors.affiliation: Genetic Engineering Team, RIKEN Center for Life Science Technologies, 2-2-3 Minatojima Minami-machi,Chuou-ku, Kobe 650-0047, Japan.
authors.forename: Takaya
authors.initials: T
authors.lastname: Abe
authors.affiliation: Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Uemura
authors.affiliation: Division of Embryology, National Institute for Basic Biology, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, Japan; Department of Basic Biology, SOKENDAI (The Graduate University for Advanced Studies), Japan; Genetic Engineering Team, RIKEN Center for Life Science Technologies, 2-2-3 Minatojima Minami-machi,Chuou-ku, Kobe 650-0047, Japan. Electronic address: fujimori@nibb.ac.jp.
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Fujimori
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000235:genetics*; D016764:Cell Polarity / Q000235:genetics; D004848:Epithelium / Q000254:growth & development / Q000378:metabolism; D005260:Female; D036681:Fluorescence Recovery After Photobleaching; D049452:Green Fluorescent Proteins / Q000235:genetics; D060588:LIM Domain Proteins / Q000235:genetics*; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D009024:Morphogenesis / Q000235:genetics*; D009419:Nerve Tissue Proteins / Q000235:genetics*; D010057:Oviducts / Q000254:growth & development / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D060588:LIM Domain Proteins; C431577:Ltap protein, mouse; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; C518732:Prickle2 protein, mouse; C528120:Vangl1 protein, mouse; D049452:Green Fluorescent Proteins
keywords: Microtubule; Oviduct; Planar cell polarity; Vangl2