Abstract
Activation of the canonical Wnt pathway leads to the transcriptional activation of a particular subset of downstream Wnt target genes. To track this localized cellular output in a living organism, reporter constructs can be designed containing multimerized consensus lymphoid enhancer binding factor (LEF)-1/T cell factor (TCF) transcription factor binding sites, generally referred to as TCF optimal promoter (TOP) sites. In Xenopus, several Wnt-responsive reporter systems have been designed containing a number of these TOP sites that, in combination with a minimal promoter, drive the expression of a reporter gene. Following transgenic integration in Xenopus embryos, a Wnt reporter tool reveals the spatiotemporal delineation of endogenous Wnt pathway activities throughout development. Assumed to be a general readout of the Wnt pathway, such reporters can assist in elucidating unknown functional implications in developing Xenopus embryos.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Logan, C. Y., Nusse, R. (2004) The Wnt signaling pathway in development and disease. Annu Rev Cell Dev Biol 20, 781–810.
Hoppler, S., Kavanagh, C. L. (2007) Wnt signaling: variety at the core. J Cell Sci 120, 385–393.
Xu, Q., Wang, Y., Dabdoub, A., et al. (2004) Vascular development in the retina and inner ear: control by Norrin and Friz-zled-4, a high-affinity ligand-receptor pair. Cell 116, 883–895.
Kazanskaya, O., Glinka, A., del BarcoBar-rantes, I., et al. (2004) R-Spondin2 is a secreted activator of Wnt/beta-catenin signaling and is required for Xenopus myogen-esis. Dev Cell 7, 525–534.
Kioussi, C., Briata, P., Baek, S. H., et al. (2002) Identification of a Wnt/Dvl/beta-Catenin → Pitx2 pathway mediating cell-type-specific proliferation during development. Cell 111, 673–685.
Easwaran, V., Pishvaian, M., Salimuddin, et al. (1999) Cross-regulation of beta-cat-enin-LEF/TCF and retinoid signaling pathways. Curr Biol 9, 1415–1418.
Zorn, A. M., Barish, G. D., Williams, B. O., et al. (1999) Regulation of Wnt signaling by Sox proteins: XSox17 alpha/beta and XSox3 physically interact with beta-catenin. Mol Cell 4, 487–498.
Barolo, S. (2006) Transgenic Wnt/TCF pathway reporters: all you need is Lef? Onco-gene 25, 7505–7511.
Korinek, V., Barker, N., Morin, P. J., et al. (1997) Constitutive transcriptional activation by a beta-catenin-Tcf complex in APC-/- colon carcinoma. Science 275, 1784–1787.
Dorsky, R. I., Sheldahl, L. C., Moon, R. T. (2002) A transgenic Lef1/beta-catenin-dependent reporter is expressed in spatially restricted domains throughout zebrafish development. Dev Biol 241, 229–237.
DasGupta, R., Fuchs, E. (1999) Multiple roles for activated LEF/TCF transcription complexes during hair follicle development and differentiation. Development 126, 4557–4568.
Staal, F.J., Meeldijk, J., Moerer, P., et al. (2001) Wnt signaling is required for thy-mocyte development and activates Tcf-1 mediated transcription. Eur J Immunol 31, 285–293.
Maretto, S., Cordenonsi, M., Dupont, S., et al. (2003) Mapping Wnt/beta-catenin signaling during mouse development and in colorectal tumors. Proc Natl Acad Sci USA 100, 3299–3304.
Mohamed, O. A., Clarke, H. J., Dufort, D. (2004) Beta-catenin signaling marks the prospective site of primitive streak formation in the mouse embryo. Dev Dyn 231, 416–424.
Nakaya, M. A., Biris, K., Tsukiyama, T., et al. (2005) Wnt3a links left-right determination with segmentation and antero-posterior axis elongation. Development 132, 5425–5436.
Geng, X., Xiao, L., Lin, G. F., et al. (2003) Lef/ Tcf-dependent Wnt/beta-catenin signaling during Xenopus axis specification. FEBS Lett 547, 1–6.
Denayer, T., Van Roy, F., Vleminckx, K. (2006) In vivo tracing of canonical Wnt signaling in Xenopus tadpoles by means of an inducible transgenic reporter tool. FEBS Lett 580, 393–398.
Sive, H. L., Grainger, R. M., Harland, R. M. (eds.) (2000) Early Development of Xenopus laevis, A Laboratory Manual. Cold Spring Harbor Laboratory Press, NY.
Staal, F. J., Burgering, B. M., van de Weter-ing, M., et al. (1999) Tcf-1-mediated transcription in T lymphocytes: differential role for glycogen synthase kinase-3 in fibroblasts and T cells. Int Immunol 11, 317–323.
Chalfie, M., Tu, Y., Euskirchen, G., et al. (1994) Green fluorescent protein as a marker for gene expression. Science 263, 802–805.
Corish, P., Tyler-Smith, C. (1999) Attenuation of green fluorescent protein half-life in mammalian cells. Protein Eng 12, 1035–1040.
Huang, W. Y., Aramburu, J., Douglas, P. S., et al. (2000) Transgenic expression of green fluorescence protein can cause dilated car-diomyopathy. Nat Med 6, 482–483.
Li, X., Zhao, X., Fang, Y., et al. (1998) Generation of destabilized green fluorescent protein as a transcription reporter. J Biol Chem 273, 34970–34975.
Bevis, B. J., Glick, B. S. (2002) Rapidly maturing variants of the Discosoma red fluorescent protein (DsRed) Nat Biotechnol 20, 83–87.
Chae, J., Zimmerman, L. B., Grainger, R. M. (2002) Inducible control of tissue-specific transgene expression in Xenopus tropicalis transgenic lines. Mech Dev 117, 235–241.
Das, B., Brown, D. D. (2004) Controlling transgene expression to study Xenopus lae-vis metamorphosis. Proc Natl Acad Sci USA 101, 4839–4842.
Recillas-Targa, F., Pikaart, M. J., Burgess-Beusse, B., et al. (2002) Position-effect protection and enhancer blocking by the chicken beta-globin insulator are separable activities. Proc Natl Acad Sci USA 99, 6883–6888.
Sekkali, B., Tran, H. T., Crabbe, E., et al. (2008) Chicken beta-globin insulator overcomes variegation of transgenes in Xenopus embryos. Faseb J 22, 2534–2540.
De Robertis, E. M., Larrain, J., Oelge-schlager, M., et al. (2000) The establishment of Spemann's organizer and patterning of the vertebrate embryo. Nat Rev Genet 1, 171–181.
Amaya, E., Kroll, K. L. (1999) A method for generating transgenic frog embryos in (Sharpe, P., Mason, I., eds.) Molecular Embryology: Methods and Protocols, Humana, Totowa, NJ, pp. 393–414.
Hirsch, N., Zimmerman, L. B., Gray, J., et al. (2002) Xenopus tropicalis transgenic lines and their use in the study of embryonic induction. Dev Dyn 225, 522–535.
Jansen, E. J., Holling, T. M., van Herp, F., et al. (2002) Transgene-driven protein expression specific to the intermediate pituitary melanotrope cells of Xenopus laevis. FEBS Lett 516, 201–207.
Nieuwkoop, P. D., Faber, J. (eds.) (1994) Normal table of Xenopus laevis (Daudin): a systematical and chronological survey of development from the fertilized egg till the end of metamorphosis. Garland Science, NY.
Broadbent, J., Read, E. M. (1999) Wholemount in situ hybridization of Xenopus and zebrafish embryos in (Guille, M., ed.) Molecular Methods in Developmental Biology, Xenopus and Zebrafish, Humana, Totowa, NJ, pp. 57–67.
Acknowledgments
TD is a postdoctoral fellow of the Research Foundation—Flanders (FWO). Research is supported by the Belgian Foundation against Cancer and the Research Foundation—Flanders (FWO).
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Denayer, T., Tran, H.T., Vleminckx, K. (2008). Transgenic Reporter Tools Tracing Endogenous Canonical Wnt Signaling in Xenopus . In: Vincan, E. (eds) Wnt Signaling. Methods in Molecular Biology, vol 469. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-469-2_24
Download citation
DOI: https://doi.org/10.1007/978-1-60327-469-2_24
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60327-468-5
Online ISBN: 978-1-60327-469-2
eBook Packages: Springer Protocols