Abstract
Planar cell polarity (PCP) pathway of Wnt signaling plays a crucial role to establish the polarization of cells during tissue development. Our recent findings using in vitro analyses have revealed that Ror2, a member of the Ror-family receptor tyrosine kinases, acts as a receptor or co-receptor for Wnt5a and plays a crucial role for Wnt5a-induced polarized cell migration through activating PCP pathway. Indeed, analyses of both Wnt5a and Ror2 mutant mice have shown that Wnt5a-Ror2 signaling is involved in establishing the PCP in epithelial tissues in vivo, indicating that in vitro analyses of polarized cell migration and PCP signaling induced by Wnt5a can be useful tools to explore putative regulators involved in Wnt/PCP pathway. Here, we introduce in vitro methods using cultured cells to monitor polarized cell migration and PCP signaling induced by Wnt5a.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Gubb, D., and Garcia-Bellido, A. (1982) A genetic analysis of the determination of cuticular polarity during development in Drosophila melanogaster. J Embryol Exp Morphol 68, 37–57.
Adler, P. N. (2002) Planar signaling and morphogenesis in Drosophila. Dev Cell 2, 525–535.
Klein, T. J., and Mlodzik, M. (2005) Planar cell polarization: an emerging model points in the right direction. Annu Rev Cell Dev Biol 21, 155–176.
Mlodzik, M. (2002) Planar cell polarization: do the same mechanisms regulate Drosophila tissue polarity and vertebrate gastrulation? Trends Genet 18, 564–571.
Fanto, M., and McNeill, H. (2004) Planar polarity from flies to vertebrates. J Cell Sci 117, 527–533.
Cadigan, K. M., and Nusse, R. (1997) Wnt signaling: a common theme in animal development. Genes Dev 11, 3286–3305.
Sokol, S. Y. (1999) Wnt signaling and dorso-ventral axis specification in vertebrates. Curr Opin Genet Dev 9, 405–410.
Moon, R. T., Campbell, R. M., Christian, J. L., McGrew, L. L., Shih, J., and Fraser, S. (1993) Xwnt-5A: a maternal Wnt that affects morphogenetic movements after overexpression in embryos of Xenopus laevis. Development 119, 97–111.
Heisenberg, C. P., Tada, M., Rauch, G. J., Saude, L., Concha, M. L., Geisler, R., Stemple, D. L., Smith, J. C., and Wilson, S. W. (2000) Silberblick/Wnt11 mediates convergent extension movements during zebrafish gastrulation. Nature 405, 76–81.
Sokol, S. (2000) A role for Wnts in morpho-genesis and tissue polarity. Nat Cell Biol 2, E124–125.
Tada, M., and Smith, J. C. (2000) Xwnt11 is a target of Xenopus Brachyury: regulation of gastrulation movements via Dishevelled, but not through the canonical Wnt pathway. Development 127, 2227–2238.
Oishi, I., Suzuki, H., Onishi, N., Takada, R., Kani, S., Ohkawara, B., Koshida, I., Suzuki, K., Yamada, G., Schwabe, G. C., Mundlos, S., Shibuya, H., Takada, S., and Minami, Y. (2003) The receptor tyrosine kinase Ror2 is involved in non-canonical Wnt5a/JNK signalling pathway. Genes Cells 8, 645–654.
Qian, D., Jones, C., Rzadzinska, A., Mark, S., Zhang, X., Steel, K. P., Dai, X., and Chen, P. (2007) Wnt5a functions in planar cell polarity regulation in mice. Dev Biol 306, 121–133.
Nomachi, A., Nishita, M., Inaba, D., Enomoto, M., Hamasaki, M., and Minami, Y. (2008) Receptor tyrosine kinase Ror2 mediates Wnt5a-induced polarized cell migration by activating c-Jun N-terminal kinase via actin-binding protein filamin A. J Biol Chem 283, 27973–27981.
Boutros, M., Paricio, N., Strutt, D. I., and Mlodzik, M. (1998) Dishevelled activates JNK and discriminates between JNK pathways in planar polarity and wingless signaling. Cell 94, 109–118.
Yamanaka, H., Moriguchi, T., Masuyama, N., Kusakabe, M., Hanafusa, H., Takada, R., Takada, S., and Nishida, E. (2002) JNK functions in the non-canonical Wnt pathway to regulate convergent extension movements in vertebrates. EMBO Rep 3, 69–75.
Derijard, B., Hibi, M., Wu, I. H., Barrett, T., Su, B., Deng, T., Karin, M., and Davis, R. J. (1994) JNK1: a protein kinase stimulated by UV light and Ha-Ras that binds and phosphorylates the c-Jun activation domain. Cell 76, 1025–1037.
Cheyette, B. N., Waxman, J. S., Miller, J. R., Takemaru, K., Sheldahl, L. C., Khlebtsova, N., Fox, E. P., Earnest, T., and Moon, R. T. (2002) Dapper, a Dishevelled-associated antagonist of beta-catenin and JNK signaling, is required for notochord formation. Dev Cell 2, 449–461.
Pukrop, T., Klemm, F., Hagemann, T., Gradl, D., Schulz, M., Siemes, S., Trumper, L., and Binder, C. (2006) Wnt 5a signaling is critical for macrophage-induced invasion of breast cancer cell lines. Proc Natl Acad Sci USA 103, 5454–5459.
Nishita, M., Itsukushima, S., Nomachi, A., Endo, M., Wang, Z., Inaba, D., Qiao, S., Takada, S., Kikuchi, A., and Minami, Y. (2010) Ror2/Frizzled complex mediates Wnt5a-induced AP-1 activation by regulating Dishevelled polymerization. Mol Cell Biol. 30, 3610–3619.
Yamamoto, S., Nishimura, O., Misaki, K., Nishita, M., Minami, Y., Yonemura, S., Tarui, H., and Sasaki, H. (2008) Cthrc1 selectively activates the planar cell polarity pathway of Wnt signaling by stabilizing the Wnt-receptor complex. Dev Cell 15, 23–36.
Kani, S., Oishi, I., Yamamoto, H., Yoda, A., Suzuki, H., Nomachi, A., Iozumi, K., Nishita, M., Kikuchi, A., Takumi, T., and Minami, Y. (2004) The receptor tyrosine kinase Ror2 associates with and is activated by casein kinase Iepsilon. J Biol Chem 279, 50102–50109.
Scliwa, M., and Honer, B. (1993) Microtubules, centrosomes and intermediate filaments in directed cell movement. Trends Cell Biol 3, 377–380.
Nobes, C. D., and Hall, A. (1999) Rho GTPases control polarity, protrusion, and adhesion during cell movement. J Cell Biol 144, 1235–1244.
Lallemand, D., Ham, J., Garbay, S., Bakiri, L., Traincard, F., Jeannequin, O., Pfarr, C. M., and Yaniv, M. (1998) Stress-activated protein kinases are negatively regulated by cell density. Embo J 17, 5615–5626.
Takada, R., Hijikata, H., Kondoh, H., and Takada, S. (2005) Analysis of combinatorial effects of Wnts and Frizzleds on beta-catenin/armadillo stabilization and Dishevelled phosphorylation. Genes Cells 10, 919–928.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Endo, M., Nishita, M., Minami, Y. (2012). Analysis of Wnt/Planar Cell Polarity Pathway in Cultured Cells. In: Turksen, K. (eds) Planar Cell Polarity. Methods in Molecular Biology, vol 839. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-510-7_16
Download citation
DOI: https://doi.org/10.1007/978-1-61779-510-7_16
Published:
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-61779-509-1
Online ISBN: 978-1-61779-510-7
eBook Packages: Springer Protocols