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.
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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).
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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 (2020). https://doi.org/10.1038/s41418-020-00632-8