Patterning and growth control by membrane-tethered Wingless (original) (raw)

References

  1. van Amerongen, R. & Nusse, R. Towards an integrated view of Wnt signaling in development. Development 136, 3205–3214 (2009)
    Article CAS PubMed Google Scholar
  2. Clevers, H. & Nusse, R. Wnt/β-catenin signaling and disease. Cell 149, 1192–1205 (2012)
    Article CAS PubMed Google Scholar
  3. Kiecker, C. & Niehrs, C. A morphogen gradient of Wnt/β -catenin signalling regulates anteroposterior neural patterning in Xenopus. Development 128, 4189–4201 (2001)
    CAS PubMed Google Scholar
  4. Zecca, M., Basler, K. & Struhl, G. Direct and long-range action of a wingless morphogen gradient. Cell 87, 833–844 (1996)
    Article CAS PubMed Google Scholar
  5. Neumann, C. J. & Cohen, S. M. Long-range action of Wingless organizes the dorsal-ventral axis of the Drosophila wing. Development 124, 871–880 (1997)
    CAS PubMed Google Scholar
  6. Swarup, S. & Verheyen, E. M. Wnt/Wingless signaling in Drosophila. Cold Spring Harb. Perspect. Biol. 4, (2012)
  7. Garcia-Bellido, A. & Merriam, J. R. Parameters of the wing imaginal disc development of Drosophila melanogaster. Dev. Biol. 24, 61–87 (1971)
    Article CAS PubMed Google Scholar
  8. Johnston, L. A. & Gallant, P. Control of growth and organ size in Drosophila. BioEssays 24, 54–64 (2002)
    Article CAS PubMed Google Scholar
  9. Martín, F. A., Herrera, S. C. & Morata, G. Cell competition, growth and size control in the Drosophila wing imaginal disc. Development 136, 3747–3756 (2009)
    Article PubMed Google Scholar
  10. Williams, J. A., Paddock, S. W. & Carroll, S. B. Pattern formation in a secondary field: a hierarchy of regulatory genes subdivides the developing Drosophila wing disc into discrete subregions. Development 117, 571–584 (1993)
    CAS PubMed Google Scholar
  11. Couso, J. P., Knust, E. & Martinez Arias, A. Serrate and wingless cooperate to induce vestigial gene expression and wing formation in Drosophila. Curr. Biol. 5, 1437–1448 (1995)
    Article CAS PubMed Google Scholar
  12. Ng, M., Diaz-Benjumea, F. J., Vincent, J. P., Wu, J. & Cohen, S. M. Specification of the wing by localized expression of wingless protein. Nature 381, 316–318 (1996)
    Article ADS CAS PubMed Google Scholar
  13. García-García, M. J., Ramain, P., Simpson, P. & Modolell, J. Different contributions of pannier and wingless to the patterning of the dorsal mesothorax of Drosophila. Development 126, 3523–3532 (1999)
    PubMed Google Scholar
  14. Martinez Arias, A. Wnts as morphogens? The view from the wing of Drosophila. Nature Rev. Mol. Cell Biol. 4, 321–325 (2003)
    Article CAS Google Scholar
  15. Couso, J. P., Bishop, S. A. & Martinez Arias, A. The wingless signalling pathway and the patterning of the wing margin in Drosophila. Development 120, 621–636 (1994)
    CAS PubMed Google Scholar
  16. Nolo, R., Abbott, L. A. & Bellen, H. J. Senseless, a Zn finger transcription factor, is necessary and sufficient for sensory organ development in Drosophila. Cell 102, 349–362 (2000)
    Article CAS PubMed Google Scholar
  17. Jafar-Nejad, H., Tien, A.-C., Acar, M. & Bellen, H. J. Senseless and Daughterless confer neuronal identity to epithelial cells in the Drosophila wing margin. Development 133, 1683–1692 (2006)
    Article CAS PubMed Google Scholar
  18. Sato, A., Kojima, T., Ui-Tei, K., Miyata, Y. & Saigo, K. Dfrizzled-3, a new Drosophila Wnt receptor, acting as an attenuator of Wingless signaling in wingless hypomorphic mutants. Development 126, 4421–4430 (1999)
    CAS PubMed Google Scholar
  19. Sivasankaran, R., Calleja, M., Morata, G. & Basler, K. The Wingless target gene Dfz3 encodes a new member of the Drosophila Frizzled family. Mech. Dev. 91, 427–431 (2000)
    Article CAS PubMed Google Scholar
  20. Giraldez, A. J. & Cohen, S. M. Wingless and Notch signaling provide cell survival cues and control cell proliferation during wing development. Development 130, 6533–6543 (2003)
    Article CAS PubMed Google Scholar
  21. Baena-López, L. A., Franch-Marro, X. & Vincent, J.-P. Wingless promotes proliferative growth in a gradient-independent manner. Sci. Signal. 2, ra60 (2009)
    Article PubMed PubMed Central Google Scholar
  22. Zecca, M. & Struhl, G. A feed-forward circuit linking wingless, fat-dachsous signaling, and the warts-hippo pathway to Drosophila wing growth. PLoS Biol. 8, e1000386 (2010)
    Article PubMed PubMed Central Google Scholar
  23. Herr, P. & Basler, K. Porcupine-mediated lipidation is required for Wnt recognition by Wls. Dev. Biol. 361, 392–402 (2012)
    Article CAS PubMed Google Scholar
  24. Baena-López, L. A., Alexandre, C., Mitchell, A., Pashakarnis, L. & Vincent, J.-P. Accelerated genome engineering in Drosophila without sequence constraints. Development 140, 4818–4825 (2013)
    Article PubMed PubMed Central Google Scholar
  25. Bergantiños, C., Corominas, M. & Serras, F. Cell death-induced regeneration in wing imaginal discs requires JNK signalling. Development 137, 1169–1179 (2010)
    Article PubMed Google Scholar
  26. Piddini, E. & Vincent, J.-P. Interpretation of the wingless gradient requires signaling-induced self-inhibition. Cell 136, 296–307 (2009)
    Article CAS PubMed Google Scholar
  27. Dubois, L., Lecourtois, M., Alexandre, C., Hirst, E. & Vincent, J. P. Regulated endocytic routing modulates wingless signaling in Drosophila embryos. Cell 105, 613–624 (2001)
    Article CAS PubMed Google Scholar
  28. Lecuit, T. et al. Two distinct mechanisms for long-range patterning by Decapentaplegic in the Drosophila wing. Nature 381, 387–393 (1996)
    Article ADS CAS PubMed Google Scholar
  29. Pérez, L. et al. Enhancer-PRE communication contributes to the expansion of gene expression domains in proliferating primordia. Development 138, 3125–3134 (2011)
    Article PubMed Google Scholar
  30. Halder, G. et al. The Vestigial and Scalloped proteins act together to directly regulate wing-specific gene expression in Drosophila. Genes Dev. 12, 3900–3909 (1998)
    Article CAS PubMed PubMed Central Google Scholar
  31. Smith-Bolton, R. K., Worley, M. I., Kanda, H. & Hariharan, I. K. Regenerative growth in Drosophila imaginal discs is regulated by Wingless and Myc. Dev. Cell 16, 797–809 (2009)
    Article CAS PubMed PubMed Central Google Scholar
  32. Colombani, J., Andersen, D. S. & Léopold, P. Secreted peptide Dilp8 coordinates Drosophila tissue growth with developmental timing. Science 336, 582–585 (2012)
    Article ADS CAS PubMed Google Scholar
  33. Garelli, A., Gontijo, A. M., Miguela, V., Caparros, E. & Dominguez, M. Imaginal discs secrete insulin-like peptide 8 to mediate plasticity of growth and maturation. Science 336, 579–582 (2012)
    Article ADS CAS PubMed Google Scholar
  34. Strigini, M. & Cohen, S. M. Wingless gradient formation in the Drosophila wing. Curr. Biol. 10, 293–300 (2000)
    Article CAS PubMed Google Scholar
  35. Giorgianni, M. W. & Mann, R. S. Establishment of medial fates along the proximodistal axis of the Drosophila leg through direct activation of dachshund by Distalless. Dev. Cell 20, 455–468 (2011)
    Article CAS PubMed PubMed Central Google Scholar
  36. Wilder, E. L. & Perrimon, N. Dual functions of wingless in the Drosophila leg imaginal disc. Development 121, 477–488 (1995)
    CAS PubMed Google Scholar
  37. Couso, J. P., Bate, M. & Martínez-Arias, A. A wingless-dependent polar coordinate system in Drosophila imaginal discs. Science 259, 484–489 (1993)
    Article ADS CAS PubMed Google Scholar
  38. Artero, R., Furlong, E. E., Beckett, K., Scott, M. P. & Baylies, M. Notch and Ras signaling pathway effector genes expressed in fusion competent and founder cells during Drosophila myogenesis. Development 130, 6257–6272 (2003)
    Article CAS PubMed Google Scholar
  39. Alexandre, C., Lecourtois, M. & Vincent, J. Wingless and Hedgehog pattern Drosophila denticle belts by regulating the production of short-range signals. Development 126, 5689–5698 (1999)
    CAS PubMed Google Scholar

Download references

Acknowledgements

This work was supported by the UK Medical Research Council (U117584268), an ERC grant (WNTEXPORT) from the European Union to JPV and a Sir Henry Wellcome post-doctoral fellowship to L.A.B-L. (082694/Z/07/Z). We are grateful to U.-M. Fiuza for discussion and A. Mitchell for help in generating the first wingless knockout allele. Discussions with G. Struhl have led to significant improvement of the manuscript. We thank colleagues listed in the Methods Summary, as well as the Developmental Studies Hybridoma Bank and the Bloomington Stock Center for providing antibodies and fly strains.

Author information

Author notes

  1. Cyrille Alexandre and Alberto Baena-Lopez: These authors contributed equally to this work.

Authors and Affiliations

  1. MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK,
    Cyrille Alexandre, Alberto Baena-Lopez & Jean-Paul Vincent

Authors

  1. Cyrille Alexandre
  2. Alberto Baena-Lopez
  3. Jean-Paul Vincent

Contributions

All the experiments were performed jointly by L.A.B.-L. and C.A. C.A., L.A.B.-L., and J.-P.V. contributed equally to the conception of the work, the interpretation of results, and manuscript preparation.

Corresponding author

Correspondence toJean-Paul Vincent.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Extended data figures and tables

Extended Data Figure 1 Engineering the wg locus to express membrane-tethered Wg.

a, Structure of the wingless locus before targeting, after targeting, and after Cre-mediated excision. The wg{KO} allele was used as a founder line for subsequent reintegration. b, Cherry expression in wing, leg and haltere imaginal discs of larvae carrying one copy of wg{KO; Cherry}. c, Cuticle preparation of a homozygous wg{KO}larva at low and high magnification (black arrow). The phenotype is identical to that of wg CX4 homozygous embryos39. d, Diagram showing the reintegration of a wild type wingless cDNA in the wg{KO} to generate wg{KO; Wg} (note presence of mini-white). e, Diagram showing the reintegration of the NRT–Wg cDNA in wg{KO}. This was achieved using either pax_-Cherry or mini-white as a genetic marker, as indicated. f, Wing of a wild-type fly. g, Wing of a wg{KO; Wg} homozygous fly. h, Overlay of the wings shown in Fig. 1b to illustrate the mild wing size reduction in NRT_–_Wg flies. i, Wing size of wg{KO; NRT_–_Wg} homozygous (n = 14) and control (w_g{KO; NRT_–_Wg}/GlaBC) flies (n = 16, ***P < 0.001). j–l, High-magnification view of the wing margin of wild-type, homozygous wg{KO; Wg}, and homozygous wg{KO; NRT_–_Wg}. They are barely distinguishable. m–o, Views of the dorsal thorax illustrate the normal arrangement of pattern elements such as microchaetes and macrochaetes in the genotypes indicated. Error bars represent s.d. Statistical significance was assessed using Student’s _t_-test.

Extended Data Figure 2 Senseless expression and growth in _wingless_-null patches surrounded by wild-type or Neurotactin–Wingless-expressing cells.

a, b, Expression of Senseless (red) is lost in patches of wingless mutant cells (GFP-negative; wg CX4 homozygotes) except in the cells located within one cell diameter of surrounding GFP-positive cells, which are wild-type cells (a) or homozygous wg{KO; NRT_–_Wg} (b) (white arrows). Mosaics were created by mitotic recombination in a way that generates approximately the same number progenitors for the two genotypes, as described in Methods. c, Example of mosaic imaginal discs generated as above to measure the growth of wingless mutant territory (wg CX4 homozygous; GFP-negative) relative to that of wild type (c) or wg{KO; NRT_–_Wg} homozygous (d) tissue. Wg and NRT–Wg, detected with anti-Wg, are shown in red. e, Outline of the territory where the surface areas were assessed. f, Quantification of the areas colonised by wingless mutant cells (GFP-negative) in the two genetic backgrounds. On average, the _wingless_-null territory was smaller in the wg{KO; NRT_–_Wg} homozygous background (n = 24) than in the wild type (n = 20, ***P < 0.001). Error bars represent s.d. Statistical significance was assessed using Student’s _t_-test.

Extended Data Figure 3 Activity of the wingless promoter during imaginal disc development.

a, Timing of key developmental stages at 25 °C. b, Developmental timing at 18 °C as it relates to the results illustrated in c. c, Permanent labelling of wg_-expressing cells and their descendants at different stages of development. Genotype was wg{KO; Gal4}, tubulin_-gal80 ts _/UAS_-Flp; Actin FRT stop FRT _Lac_Z so that the stop cassette was only excised in cells that express wingless at the time of shifting to 29 °C to activate Flp expression and hence excision of the stop cassette. Discs were shifted from 18 °C to 29 °C at different stages (shown in b) but they were fixed and stained at the same stage, just before puparation.

Extended Data Figure 4 Tissue-specific allele switching to determine the anatomical origin of organismal developmental delay in Neurotactin–Wingless animals.

a, Cumulative pattern of _vestigial_-gal4 activity in various organs precursors. Expression of _vestigial_-gal4 at any stage or place leads to excision of the stop cassette in Actin FRT stop FRT LacZ thus marking permanently the corresponding cells. As expected, nearly the whole wing and haltere discs were labelled at the end of larval development. In wing imaginal discs, only a few cells were β-Galactosidase-negative that did not overlap with the domain of Wg expression (anti-Wg, red). In the eye antennal disc, the patterns of Wg (white arrowhead) and β-Galactosidase expression are also non-overlapping. Therefore, in combination with _UAS_-_Flp, vestigial_-gal4 is expected to excise an FRT cassette throughout the domain of wingless expression. Examination of the brain and CNS shows that _vestigial_-gal4 is unexpectedly active in these tissues. b, In larvae of genotype vestigial_-gal4, UAS_-Flp, wg{FRT Wg FRT NRT_–_Wg}/Cyo wg, most of the _wg_-expressing cells in leg, haltere and wing imaginal discs, but not in the brain and CNS, were converted to expressing NRT-HA-Wg (anti-HA; green). c, Developmental timing in wg{KO; WT Body ; NRT_–_Wg Disc} (_vestigial_-gal4, UAS_-Flp, wg{FRT Wg FRT NRT_–_Wg}) and control (vestigial_-gal4, UAS_-Flp, wg{FRT Wg FRT NRT_–_Wg}/GlaBC) larvae (80 animals, 4 experiments). The two data sets cannot be statistically distinguished (P > 0.05). d, Adult wing size for three genotypes: wg{KO; WT Body ; NRT–Wg Discs}/GlaB_c obtained from selfed vestigial_-gal4, UAS_-Flp, wg{FRT Wg FRT NRT_–_Wg}/GlaBc (n = 16, shown in black); wg{KO; WT Body ; NRT_–_Wg Discs}, obtained from homozygous _vestigial_-_gal4, UAS_-Flp, wg{FRT Wg FRT NRT_–_Wg} (n = 15, shown in purple); and wg{KO; NRT–Wg Body ;WT Discs}, obtained from homozygous _vestigial_-_gal4, UAS_-Flp, wg{FRT NRT_–_Wg FRT Wg} (n = 13, shown in grey). e–g, Extent of Distal-less expression in wg{KO; WT Body ; NRT_–_Wg Discs} heterozygotes (over Cyo; e) and homozygotes (f). All the discs were obtained from immobile larvae at the time of anterior spiracle eversion, an event that marks the onset of pupariation. The extent of the Distal-less domain was estimated from the surface area of a polygon drawn around the zone of immunoreactivity, as shown. The results, plotted in panel g, show a mild reduction in wg{KO; WT Body ; NRT_–_Wg Discs}discs (n = 13) compared to controls (n = 20; ***P < 0.001; n.s., not significantly different). Error bars represent s.d. Statistical significance was assessed using Student’s _t_-test.

Supplementary information

PowerPoint slides

Rights and permissions

About this article

Cite this article

Alexandre, C., Baena-Lopez, A. & Vincent, JP. Patterning and growth control by membrane-tethered Wingless.Nature 505, 180–185 (2014). https://doi.org/10.1038/nature12879

Download citation