Abstracts
Wingless (Wg)/Wnt family proteins are essential for animal development and adult homeostasis. Drosophila Wg secreted from the dorsal-ventral (DV) midline in wing discs forms a concentration gradient that is shaped by diffusion rate and stability of Wg. To understand how the gradient of extracellular Wg is generated, we compared the secretion route of NRT-Wg, an artificial membrane-tethered form of Wg that is supposedly not secreted but still supports fly development, to that of wild-type Wg. We found that wild-type Wg is secreted by both conventional Golgi transport and via extracellular vesicles (EVs), and NRT-Wg can be also secreted via EVs. Furthermore, wild-type Wg secreted by Golgi transport diffused and formed Wg gradient but Wg-containing EVs did not diffuse at all. In case of Wg stability, Sol narae (Sona), a metalloprotease that cleaves Wg, contributes to generate a steep Wg gradient. Interestingly, Wg was also produced in the presumptive wing blade region, which indicates that NRT-Wg on EVs expressed in the blade allows the blade cells to proliferate and differentiate without Wg diffused from the DV midline. We propose that EV-associated Wg induces Wg signaling in autocrine and juxtaposed manners whereas Wg secreted by Golgi transport forms gradient and acts in the long-range signaling, and different organs differentially utilize these two types of Wg signaling for their own development.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Teo JL, Kahn M. The Wnt signaling pathway in cellular proliferation and differentiation: a tale of two coactivators. Adv Drug Deliv Rev. 2010;62:1149–55.
Steinhart Z, Angers S. Wnt signaling in development and tissue homeostasis. Development. 2018;145:dev146589.
Masckauchan TN, Shawber CJ, Funahashi Y, Li CM, Kitajewski J. Wnt/beta-catenin signaling induces proliferation, survival and interleukin-8 in human endothelial cells. Angiogenesis. 2005;8:43–51.
Humphries AC, Mlodzik M. From instruction to output: Wnt/PCP signaling in development and cancer. Curr Opin Cell Biol. 2018;51:110–6.
Nusse R, Fuerer C, Ching W, Harnish K, Logan C, Zeng A, et al. Wnt signaling and stem cell control. Cold Spring Harb Symp Quant Biol. 2008;73:59–66.
Logan CY, Nusse R. The Wnt signaling pathway in development and disease. Annu Rev Cell Dev Biol. 2004;20:781–810.
Kiecker C, Niehrs C. A morphogen gradient of Wnt/beta-catenin signalling regulates anteroposterior neural patterning in Xenopus. Development. 2001;128:4189–201.
Yamaguchi TP. Heads or tails: Wnts and anterior-posterior patterning. Curr Biol. 2001;11:R713–724.
Petersen CP, Reddien PW. Wnt signaling and the polarity of the primary body axis. Cell. 2009;139:1056–68.
Simons M, Mlodzik M. Planar cell polarity signaling: from fly development to human disease. Annu Rev Genet. 2008;42:517–40.
Sharma RP. Wingless - a new mutant in D. melanogaster. Drosoph Inf Serv. 1973;50:134.
Nusse R, Varmus HE. Many tumors induced by the mouse mammary tumor virus contain a provirus integrated in the same region of the host genome. Cell. 1982;31:99–109.
Clevers H. Wnt/beta-catenin signaling in development and disease. Cell. 2006;127:469–80.
Mann MC. The occurrence and hereditary behavior of two new dominant mutations in an inbred strain of Drosophila melanogaster. Genetics. 1923;8:27–36.
Strigini M, Cohen SM. Wingless gradient formation in the Drosophila wing. Curr Biol. 2000;10:293–300.
Delanoue R, Legent K, Godefroy N, Flagiello D, Dutriaux A, Vaudin P, et al. The Drosophila wing differentiation factor Vestigial-Scalloped is required for cell proliferation and cell survival at the dorso-ventral boundary of the wing imaginal disc. Cell Death Differ. 2004;11:110–22.
Neumann CJ, Cohen SM. A hierarchy of cross-regulation involving Notch, wingless, vestigial and cut organizes the dorsal/ventral axis of the Drosophila wing. Development. 1996;122:3477–85.
Zecca M, Basler K, Struhl G. Direct and long-range action of a wingless morphogen gradient. Cell. 1996;87:833–44.
Alexandre C, Baena-Lopez A, Vincent JP. Patterning and growth control by membrane-tethered Wingless. Nature. 2014;505:180–5.
Tian A, Duwadi D, Benchabane H, Ahmed Y. Essential long-range action of Wingless/Wnt in adult intestinal compartmentalization. PLoS Genet. 2019;15:e1008111.
Wartlick O, Kicheva A, Gonzalez-Gaitan M. Morphogen gradient formation. Cold Spring Harb Perspect Biol. 2009;1:a001255.
Won JH, Kim GW, Kim JY, Cho DG, Kwon B, Bae YK, et al. ADAMTS Sol narae cleaves extracellular Wingless to generate a novel active form that regulates cell proliferation in Drosophila. Cell Death Dis. 2019;10:564.
Janda CY, Waghray D, Levin AM, Thomas C, Garcia KC. Structural basis of Wnt recognition by Frizzled. Science. 2012;337:59–64.
Bazan JF, Janda CY, Garcia KC. Structural architecture and functional evolution of Wnts. Dev Cell. 2012;23:227–32.
von Maltzahn J, Zinoviev R, Chang NC, Bentzinger CF, Rudnicki MA. A truncated Wnt7a retains full biological activity in skeletal muscle. Nat Commun. 2013;4:2869.
Port F, Basler K. Wnt Trafficking: new Insights into Wnt Maturation, Secretion and Spreading. Traffic. 2010;11:1265–71.
Gross JC, Chaudhary V, Bartscherer K, Boutros M. Active Wnt proteins are secreted on exosomes. Nat Cell Biol. 2012;14:1036–45.
Gross JC, Boutros M. Secretion and extracellular space travel of Wnt proteins. Curr Opin Genet Dev. 2013;23:385–90.
Thery C, Zitvogel L, Amigorena S. Exosomes: composition, biogenesis and function. Nat Rev Immunol. 2002;2:569–79.
Thery C, Ostrowski M, Segura E. Membrane vesicles as conveyors of immune responses. Nat Rev Immunol. 2009;9:581–93.
Lee SS, Won JH, Lim GJ, Han J, Lee JY, Cho KO, et al. A novel population of extracellular vesicles smaller than exosomes promotes cell proliferation. Cell Commun Signal. 2019;17:95.
Beckett K, Monier S, Palmer L, Alexandre C, Green H, Bonneil E, et al. Drosophila S2 cells secrete wingless on exosome-like vesicles but the wingless gradient forms independently of exosomes. Traffic. 2013;14:82–96.
Panakova D, Sprong H, Marois E, Thiele C, Eaton S. Lipoprotein particles are required for Hedgehog and Wingless signalling. Nature. 2005;435:58–65.
Barthalay Y, Hipeau-Jacquotte R, de la Escalera S, Jimenez F, Piovant M. Drosophila neurotactin mediates heterophilic cell adhesion. EMBO J. 1990;9:3603–9.
de la Escalera S, Bockamp EO, Moya F, Piovant M, Jimenez F. Characterization and gene cloning of neurotactin, a Drosophila transmembrane protein related to cholinesterases. EMBO J. 1990;9:3593–601.
Chaudhary V, Hingole S, Frei J, Port F, Strutt D, Boutros M. Robust Wnt signaling is maintained by a Wg protein gradient and Fz2 receptor activity in the developing Drosophila wing. Development. 2019;146:dev174789.
Katanaev VL, Solis GP, Hausmann G, Buestorf S, Katanayeva N, Schrock Y, et al. Reggie-1/flotillin-2 promotes secretion of the long-range signalling forms of Wingless and Hedgehog in Drosophila. EMBO J. 2008;27:509–21.
Neumann CJ, Cohen SM. Long-range action of Wingless organizes the dorsal-ventral axis of the Drosophila wing. Development. 1997;124:871–80.
Lecuit T, Cohen SM. Proximal-distal axis formation in the Drosophila leg. Nature. 1997;388:139–45.
Kim GW, Won JH, Lee OK, Lee SS, Han JH, Tsogtbaatar O, et al. Sol narae (Sona) is a Drosophila ADAMTS involved in Wg signaling. Sci Rep. 2016;6:31863.
Couso JP, Bate M, Martinez-Arias A. A wingless-dependent polar coordinate system in Drosophila imaginal discs. Science. 1993;259:484–9.
Ng M, Diaz-Benjumea FJ, Vincent JP, Wu J, Cohen SM. Specification of the wing by localized expression of wingless protein. Nature. 1996;381:316–8.
Blair SS. A role for the segment polarity gene shaggy-zeste white 3 in the specification of regional identity in the developing wing of Drosophila. Dev Biol. 1994;162:229–44.
Phillips RG, Whittle JR. Wingless expression mediates determination of peripheral nervous system elements in late stages of Drosophila wing disc development. Development. 1993;118:427–38.
Giraldez AJ, Copley RR, Cohen SM. HSPG modification by the secreted enzyme Notum shapes the Wingless morphogen gradient. Dev Cell. 2002;2:667–76.
Evans CJ, Olson JM, Ngo KT, Kim E, Lee NE, Kuoy E, et al. G-TRACE: rapid Gal4-based cell lineage analysis in Drosophila. Nat Methods. 2009;6:603–5.
Maves L, Schubiger G. A molecular basis for transdetermination in Drosophila imaginal discs: interactions between wingless and decapentaplegic signaling. Development. 1998;125:115–24.
Klein T, Arias AM. Different spatial and temporal interactions between Notch, wingless, and vestigial specify proximal and distal pattern elements of the wing in Drosophila. Dev Biol. 1998;194:196–212.
Kooh PJ, Fehon RG, Muskavitch MA. Implications of dynamic patterns of Delta and Notch expression for cellular interactions during Drosophila development. Development. 1993;117:493–507.
Zhang Y, Liu Y, Liu H, Tang WH. Exosomes: biogenesis, biologic function and clinical potential. Cell Biosci. 2019;9:19.
Zhang ZG, Buller B, Chopp M. Exosomes - beyond stem cells for restorative therapy in stroke and neurological injury. Nat Rev Neurol. 2019;15:193–203.
Baena-Lopez LA, Franch-Marro X, Vincent JP. Wingless promotes proliferative growth in a gradient-independent manner. Sci Signal. 2009;2:ra60.
Ekstrom EJ, Bergenfelz C, von Bulow V, Serifler F, Carlemalm E, Jonsson G, et al. WNT5A induces release of exosomes containing pro-angiogenic and immunosuppressive factors from malignant melanoma cells. Mol Cancer. 2014;13:88.
Hooper C, Sainz-Fuertes R, Lynham S, Hye A, Killick R, Warley A, et al. Wnt3a induces exosome secretion from primary cultured rat microglia. BMC Neurosci. 2012;13:144.
Farin HF, Jordens I, Mosa MH, Basak O, Korving J, Tauriello DV, et al. Visualization of a short-range Wnt gradient in the intestinal stem-cell niche. Nature. 2016;530:340–3.
Yamamoto H, Awada C, Hanaki H, Sakane H, Tsujimoto I, Takahashi Y, et al. The apical and basolateral secretion of Wnt11 and Wnt3a in polarized epithelial cells is regulated by different mechanisms. J Cell Sci. 2013;126:2931–43.
Beaven R, Denholm B. Release and spread of Wingless is required to pattern the proximo-distal axis of Drosophila renal tubules. Elife. 2018;7:e35373.
Apitz H, Salecker I. Spatio-temporal relays control layer identity of direction-selective neuron subtypes in Drosophila. Nat Commun. 2018;9:2295.
Vincent JP, Magee T. Tiki casts a spell on Wnt. Cell. 2012;149:1426–7.
Sanchez-Pulido L, Ponting CP. Tiki, at the head of a new superfamily of enzymes. Bioinformatics. 2013;29:2371–4.
Zhang X, Abreu JG, Yokota C, MacDonald BT, Singh S, Coburn KL, et al. Tiki1 is required for head formation via Wnt cleavage-oxidation and inactivation. Cell. 2012;149:1565–77.
Zhang X, MacDonald BT, Gao H, Shamashkin M, Coyle AJ, Martinez RV, et al. Characterization of Tiki, a New Family of Wnt-specific Metalloproteases. J Biol Chem. 2016;291:2435–43.
Klein T. Wing disc development in the fly: the early stages. Curr Opin Genet Dev. 2001;11:470–5.
Pfeiffer S, Ricardo S, Manneville JB, Alexandre C, Vincent JP. Producing cells retain and recycle Wingless in Drosophila embryos. Curr Biol. 2002;12:957–62.
Baena-Lopez LA, Alexandre C, Mitchell A, Pasakarnis L, Vincent JP. Accelerated homologous recombination and subsequent genome modification in Drosophila. Development. 2013;140:4818–25.
Prasad M, Bajpai R, Shashidhara LS. Regulation of Wingless and Vestigial expression in wing and haltere discs of Drosophila. Development. 2003;130:1537–47.
Pignoni F, Zipursky SL. Induction of Drosophila eye development by decapentaplegic. Development. 1997;124:271–8.
Acknowledgements
We thank J. Jung and J.Y. Kim for their critical reading of this paper. We are in debt to J. P. Vincent, K. Basler, S. Eaton, S. M. Cohen, L. S. Shashidhara, R. Holmgren for fly lines, Gary Struhl, J.W. Kim, K. Basler, and J. P. Vincent for DNA constructs, and two reviewers who helped to improve this report. We thank Bloomington Stock Center, Drosophila Genetic Resource Center, Korea Drosophila Resource Center, and Developmental Studies Hybridoma Bank for fly strains and antibodies. This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2017R1A2B4009254, 2019R1H1A2039726, and 2019R1A6A1A10073887).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Edited by E. Baehrecke
Rights and permissions
About this article
Cite this article
Won, JH., Cho, KO. Wg secreted by conventional Golgi transport diffuses and forms Wg gradient whereas Wg tethered to extracellular vesicles do not diffuse. Cell Death Differ 28, 1013–1025 (2021). https://doi.org/10.1038/s41418-020-00632-8
Received:
Revised:
Accepted:
Published:
Version of record:
Issue date:
DOI: https://doi.org/10.1038/s41418-020-00632-8
