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Cooperation of Syd-1 with Neurexin synchronizes pre- with postsynaptic assembly

Nature Neuroscience volume 15, pages 1219–1226 (2012)Cite this article

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Abstract

Synapse formation and maturation requires bidirectional communication across the synaptic cleft. The trans-synaptic Neurexin-Neuroligin complex can bridge this cleft, and severe synapse assembly deficits are found in Drosophila melanogaster neuroligin (Nlg1, dnlg1) and neurexin (Nrx-1, dnrx) mutants. We show that the presynaptic active zone protein Syd-1 interacts with Nrx-1 to control synapse formation at the Drosophila neuromuscular junction. Mutants in Syd-1 (RhoGAP100F, dsyd-1), Nrx-1 and Nlg1 shared active zone cytomatrix defects, which were nonadditive. Syd-1 and Nrx-1 formed a complex in vivo, and Syd-1 was important for synaptic clustering and immobilization of Nrx-1. Consequently, postsynaptic clustering of Nlg1 was affected in Syd-1 mutants, and in vivo glutamate receptor incorporation was changed in Syd-1, Nrx-1 and Nlg1 mutants. Stabilization of nascent Syd-1–Liprin-α (DLiprin-α) clusters, important to initialize active zone formation, was Nlg1 dependent. Thus, cooperation between Syd-1 and Nrx-1–Nlg1 seems to orchestrate early assembly processes between pre- and postsynaptic membranes, promoting avidity of newly forming synaptic scaffolds.

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Figure 1: Double mutants analysis of Nrx-1, Nlg1 and Syd-1.
Figure 2: Comparison of active zone ultrastructure for Nrx-1, Nlg1 and Syd-1 mutants.
Figure 3: Reduced endogenous Nrx-1 and Nlg1 clusters at Syd-1 mutant NMJs.
Figure 4: Biochemical and cell biological analysis of Syd-1–Nrx-1 complex.
Figure 5: Syd-1 influences localization and FRAP of Nrx-1 at NMJ.
Figure 6: In vivo imaging of Liprin-α–Syd-1 cluster dynamics at Nlg1 mutant NMJs.
Figure 7: Analysis of in vivo glutamate receptor incorporation in Nlg1, Nrx-1 and Syd-1 mutants.

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Acknowledgements

We would like to thank C. Quentin and A. Stawrakakis for excellent technical assistance and R. Jahn for the use of equipment. T. Andlauer and U. Thomas critically read the manuscript. We further thank M. Bhat (University of North Carolina) and W. Xie (Southeast University, Nanjing) for generously sharing reagents. This work was supported by grants from the Deutsche Forschungsgemeinschaft to S.J.S. (Exc 257, SI849/2-1 and 2-2, TP A16/SFB 551, TP B23/SFB581, TP A3 and A6/SFB 958) and H.A. (AB 116/3-2). M.H. is funded by a European Research Council Starting Grant (Astrofunc).

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Author notes

  1. David Owald, Omid Khorramshahi and Varun K Gupta: These authors contributed equally to this work.

Authors and Affiliations

  1. Freie Universität Berlin, Institute for Biology/Genetics, Berlin, Germany

    David Owald, Omid Khorramshahi, Varun K Gupta, Harald Depner, Wernher Fouquet, Carolin Wichmann, Sara Mertel, Eric Reynolds, Matthew Holt & Stephan J Sigrist

  2. NeuroCure, Charité, Berlin, Germany

    David Owald, Omid Khorramshahi, Varun K Gupta, Harald Depner, Wernher Fouquet, Carolin Wichmann, Sara Mertel, Eric Reynolds & Stephan J Sigrist

  3. University of Münster, Institute for Neurobiology, Münster, Germany

    Daniel Banovic & Hermann Aberle

  4. Heinrich Heine University Düsseldorf, Functional Cell Morphology Lab, Düsseldorf, Germany

    Daniel Banovic & Hermann Aberle

  5. European Neuroscience Institute, Göttingen, Germany

    Stefan Eimer

  6. VIB Center for the Biology of Disease, Leuven, Belgium

    Matthew Holt

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Contributions

D.O., O.K. and S.J.S. designed research. D.O., O.K., V.K.G., D.B., H.D., W.F., C.W., S.M. and E.R. performed experiments. D.O., O.K., V.K.G., D.B., H.D., W.F., C.W., S.M., H.A. and S.J.S. analyzed data. M.H. and S.E. shared protocols, reagents and advice. All authors commented on the paper. D.O., O.K. and S.J.S. wrote the paper.

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Correspondence to Stephan J Sigrist.

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Owald, D., Khorramshahi, O., Gupta, V. et al. Cooperation of Syd-1 with Neurexin synchronizes pre- with postsynaptic assembly. Nat Neurosci 15, 1219–1226 (2012). https://doi.org/10.1038/nn.3183

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