TY - JOUR
T1 - Dynamics of planar cell polarity protein Vangl2 in the mouse oviduct epithelium
AU - Shi, Dongbo
AU - Usami, Fumiko
AU - Komatsu, Kouji
AU - Oka, Sanae
AU - Abe, Takaya
AU - Uemura, Tadashi
AU - Fujimori, Toshihiko
N1 - Funding Information:
We thank I. Nakagawa for DNA construction; T. Sato for providing glutathione S-transferase expression vector; E. Fuchs for the plasmid of Celsr1-EGFP and of the plasmid of Celsr1 antigen; M. Takeichi for ECCD-2 hybridoma; S. Aizawa for critical comments on the generation of transgenic mice; T. F. Day for critical readings of our manuscript. We thank Spectrography and Bioimaging Facility, Functional Genomics Facility, NIBB Core Research Facilities for technical support. This work was supported by a CREST grant to T.U. and T.F, and by a grant of The Mitsubishi Foundation to T.U. and T.F. D.S. was a Research Fellow of the Japan Society for the Promotion of Science.
Publisher Copyright:
© 2016 Elsevier B.V.
PY - 2016/8/1
Y1 - 2016/8/1
N2 - 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.
AB - 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.
KW - Microtubule
KW - Oviduct
KW - Planar cell polarity
KW - Vangl2
UR - http://www.scopus.com/inward/record.url?scp=84975488373&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84975488373&partnerID=8YFLogxK
U2 - 10.1016/j.mod.2016.05.002
DO - 10.1016/j.mod.2016.05.002
M3 - Article
C2 - 27155041
AN - SCOPUS:84975488373
SN - 0925-4773
VL - 141
SP - 78
EP - 89
JO - Cell Differentiation and Development
JF - Cell Differentiation and Development
ER -