Abstract
Neural circuit assembly requires precise dendrite and axon targeting. We identified an evolutionarily conserved endoplasmic reticulum (ER) protein, Meigo, from a mosaic genetic screen in Drosophila melanogaster. Meigo was cell-autonomously required in olfactory receptor neurons and projection neurons to target their axons and dendrites to the lateral antennal lobe and to refine projection neuron dendrites into individual glomeruli. Loss of Meigo induced an unfolded protein response and reduced the amount of neuronal cell surface proteins, including Ephrin. Ephrin overexpression specifically suppressed the projection neuron dendrite refinement defect present in meigo mutant flies, and ephrin knockdown caused a similar projection neuron dendrite refinement defect. Meigo positively regulated the level of Ephrin N-glycosylation, which was required for its optimal function in vivo. Thus, Meigo, an ER-resident protein, governs neuronal targeting specificity by regulating ER folding capacity and protein N-glycosylation. Furthermore, Ephrin appears to be an important substrate that mediates Meigo's function in refinement of glomerular targeting.
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Acknowledgements
We thank J.B. Thomas, Bloomington and the Kyoto Drosophila Stock Center for fly stocks, A.H. Brand (University of Cambridge) for the Ephrin antibody and fly stocks, S. Goto (Rikkyo University) for the dGLG1 (120 kDa) antibody, C. Field (Harvard University) for the Lava Lamp antibody, T. Uemura (Kyoto University) for the UAS-DNcadherin plasmid, H.D. Ryoo (New York University) and P.M. Domingos (Instituto de Tecnologia Química e Biológica) for xbp1:EGFP-related reagents and the Hsc3 antibody, C.-H. Chen for advice on constructing shRNAs, G. Thomas and M. Khanna for advice on performing OptiPrep density gradient centrifugation, and all of the members of the Miura and Luo laboratories for comments on this study. We especially thank T. Mosca for improving the manuscript and M. Okumura and T.C. for the blind test. L.L. is funded by the Howard Hughes Medical Institute. This work was supported by grants from the Ministry of Education, Culture, Sports, Science and Technology in Japan to M.M. and T.C., the Japan Society for the Promotion of Science to S.U.S., M.M. and T.C., the Japan Science and Technology Agency to M.M. and T.C., and the US National Institutes of Health (R01 DC005982) to L.L.
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S.U.S. performed most of the experiments and analyzed the data. S.H., K.C. and T.K. assisted in some experiments. T.C. supervised the project. S.U.S. and T.C. wrote the paper with feedback from L.L. and M.M.
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Sekine, S., Haraguchi, S., Chao, K. et al. Meigo governs dendrite targeting specificity by modulating Ephrin level and N-glycosylation. Nat Neurosci 16, 683–691 (2013). https://doi.org/10.1038/nn.3389
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DOI: https://doi.org/10.1038/nn.3389