Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Neuronal synapse interaction reconstituted between live cells and supported lipid bilayers

Nature Chemical Biology volume 1, pages 283–289 (2005)Cite this article

Abstract

In the nervous system, homophilic and heterophilic adhesion molecules participate in the induction and differentiation of presynaptic transmitter release sites. We focus on the heterophilic interaction between postsynaptic neuroligin-1 (Nlg) and presynaptic β-neurexin (Nrx). Nlg has previously been shown to trigger presynaptic differentiation in a Nrx-expressing axon even when presented on a non-neuronal cell or on beads coated with lipid bilayers. We have now developed a new method to measure single molecule and ensemble distribution of Nrx and Nlg at the contact site between a non-neuronal Nrx-expressing cell and a flat supported glycosylphosphoinositol–neuroligin-1 (GPI-Nlg) lipid bilayer and relate them to adhesion as measured by cell migration and gravity dissociation. We find that within minutes after cell-bilayer contact, Nrx accumulates at the contact site and the contact area is expanded. The strength of cell-bilayer adhesion depends on the morphology of Nrx accumulation, with the focal concentration strengthening adhesion. The results suggest that Nlg-Nrx interaction rapidly establishes a weak, but specific, adhesion between dynamic pre- and postsynaptic processes, which may ultimately require additional molecules for synapse stabilization.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Reconstitution of GPI-Nlg in a fluid-supported lipid bilayer.
Figure 2: Distribution of GFP-Nrx in a HEK293 cell in contact with GPI-Nlg–supported lipid bilayer.
Figure 3: Cell immobilization associated with formation of Nrx-Nlg puncta.
Figure 4: Correlation between distribution of GFP-Nrx in cells and cell immobilization.
Figure 5: Inversion assay: relation between Nrx-Nlg distribution and adhesion strength.

Similar content being viewed by others

References

  1. Yamagata, M., Sanes, J.R. & Weiner, J.A. Synaptic adhesion molecules. Curr. Opin. Cell Biol. 15, 621–632 (2003).

    Article  CAS  Google Scholar 

  2. Scheiffele, P. Cell-cell signaling during synapse formation in the CNS. Annu. Rev. Neurosci. 26, 485–508 (2003).

    Article  CAS  Google Scholar 

  3. Garner, C.C., Nash, J. & Huganir, R.L. PDZ domains in synapse assembly and signalling. Trends Cell Biol. 10, 274–280 (2000).

    Article  CAS  Google Scholar 

  4. Dityatev, A. et al. Polysialylated neural cell adhesion molecule promotes remodeling and formation of hippocampal synapses. J. Neurosci. 24, 9372–9382 (2004).

    Article  CAS  Google Scholar 

  5. Sytnyk, V., Leshchyns'ka, I., Dityatev, A. & Schachner, M. Trans-Golgi network delivery of synaptic proteins in synaptogenesis. J. Cell Sci. 117, 381–388 (2004).

    Article  CAS  Google Scholar 

  6. Shapiro, L. & Colman, D.R. Structural biology of cadherins in the nervous system. Curr. Opin. Neurobiol. 8, 593–599 (1998).

    Article  CAS  Google Scholar 

  7. Benson, D.L. & Tanaka, H. N-cadherin redistribution during synaptogenesis in hippocampal neurons. J. Neurosci. 18, 6892–6904 (1998).

    Article  CAS  Google Scholar 

  8. Biederer, T. et al. SynCAM, a synaptic adhesion molecule that drives synapse assembly. Science 297, 1525–1531 (2002).

    Article  CAS  Google Scholar 

  9. Ichtchenko, K. et al. Neuroligin-1—a splice site–specific ligand for β-neurexins. Cell 81, 435–443 (1995).

    Article  CAS  Google Scholar 

  10. Nguyen, T. & Sudhof, T.C. Binding properties of neuroligin 1 and neurexin 1β reveal function as heterophilic cell adhesion molecules. J. Biol. Chem. 272, 26032–26039 (1997).

    Article  CAS  Google Scholar 

  11. Song, J.Y., Ichtchenko, K., Sudhof, T.C. & Brose, N. Neuroligin 1 is a postsynaptic cell-adhesion molecule of excitatory synapses. Proc. Natl. Acad. Sci. USA 96, 1100–1105 (1999).

    Article  CAS  Google Scholar 

  12. Scheiffele, P., Fan, J.H., Choih, J., Fetter, R. & Serafini, T. Neuroligin expressed in nonneuronal cells triggers presynaptic development in contacting axons. Cell 101, 657–669 (2000).

    Article  CAS  Google Scholar 

  13. Dean, C. et al. Neurexin mediates the assembly of presynaptic terminals. Nat. Neurosci. 6, 708–716 (2003).

    Article  CAS  Google Scholar 

  14. Graf, E.R., Zhang, X., Jin, S.X., Linhoff, M.W. & Craig, A.M. Neurexins induce differentiation of GABA and glutamate postsynaptic specializations via neuroligins. Cell 119, 1013–1026 (2004).

    Article  CAS  Google Scholar 

  15. Nam, C.I. & Chen, L. Postsynaptic assembly induced by neurexin-neuroligin interaction and neurotransmitter. Proc. Natl. Acad. Sci. USA 102, 6137–6142 (2005).

    Article  CAS  Google Scholar 

  16. Axelrod, D. Total internal reflection fluorescence microscopy in cell biology. Traffic 2, 764–774 (2001).

    Article  CAS  Google Scholar 

  17. Sonnleitner, A., Mannuzzu, L.M., Terakawa, S. & Isacoff, E.Y. Structural rearrangements in single ion channels detected optically in living cells. Proc. Natl. Acad. Sci. USA 99, 12759–12764 (2002).

    Article  CAS  Google Scholar 

  18. Groves, J.T. & Dustin, M.L. Supported planar bilayers in studies on immune cell adhesion and communication. J. Immunol. Methods 278, 19–32 (2003).

    Article  CAS  Google Scholar 

  19. Sapuri, A.R., Baksh, M.M. & Groves, J.T. Electrostatically targeted intermembrane lipid exchange with micropatterned supported membranes. Langmuir 19, 1606–1610 (2003).

    Article  CAS  Google Scholar 

  20. Lee, G.M., Ishihara, A. & Jacobson, K.A. Direct observation of brownian motion of lipids in a membrane. Proc. Natl. Acad. Sci. USA 88, 6274–6278 (1991).

    Article  CAS  Google Scholar 

  21. Fein, M. et al. Lateral mobility of lipid analogues and GPI-anchored proteins in supported bilayers determined by fluorescent bead tracking. J. Membr. Biol. 135, 83–92 (1993).

    Article  CAS  Google Scholar 

  22. Hovis, J.S. & Boxer, S.G. Patterning and composition arrays of supported lipid bilayers by microcontact printing. Langmuir 17, 3400–3405 (2001).

    Article  CAS  Google Scholar 

  23. Dustin, M.L. Role of adhesion molecules in activation signaling in T lymphocytes. J. Clin. Immunol. 21, 258–263 (2001).

    Article  CAS  Google Scholar 

  24. Pierres, A., Benoliel, A.M. & Bongrand, P. Use of a laminar flow chamber to study the rate of bond formation and dissociation between surface-bound adhesion molecules: effect of applied force and distance between surfaces. Faraday Discuss. 111, 321–330 (1998).

    Article  CAS  Google Scholar 

  25. McClay, D.R., Wessel, G.M. & Marchase, R.B. Inter-cellular recognition—quantitation of initial binding events. Proc. Natl. Acad. Sci. USA 78, 4975–4979 (1981).

    Article  CAS  Google Scholar 

  26. Westcott, K.R. & Hill, R.L. Reconstitution of a porcine submaxillary gland β-D-galactoside α2—3 sialyltransferase into liposomes. J. Biol. Chem. 260, 13116–13121 (1985).

    CAS  PubMed  Google Scholar 

  27. Hinterdorfer, P., Baber, G. & Tamm, L.K. Reconstitution of membrane-fusion sites—a total-internal-reflection fluorescence microscopy study of influenza hemagglutinin-nediated membrane-fusion. J. Biol. Chem. 269, 20360–20368 (1994).

    CAS  PubMed  Google Scholar 

  28. Angrand, M., Briolay, A., Ronzon, F. & Roux, B. Detergent-mediated reconstitution of a glycosyl-phosphatidylinositol-protein into liposomes. Eur. J. Biochem. 250, 168–176 (1997).

    Article  CAS  Google Scholar 

  29. Sackmann, E. Supported membranes: scientific and practical applications. Science 271, 43–48 (1996).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank C. Dean and P. Scheiffele for the GPI-Nlg construct, C. Dean for help with initial protein preparation, A. Pralle for help with the TIRF microscopy, H. Aaron for help with the confocal microscopy, and C. Dean, P. Scheiffele, E. Douglas, M. Forstner and S. Rozovsky for fruitful discussions. This work was supported by the Chemical Sciences, Geosciences and Biosciences Division, Office of Basic Energy Sciences, US Department of Energy (contract No. DE-AC03-76SF00098).

Author information

Authors and Affiliations

  1. Materials Sciences and Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, 94720, California, USA

    Sophie Pautot, Ehud Y Isacoff & Jay T Groves

  2. Department of Molecular & Cell Biology, University of California Berkeley, Berkeley, 94720, California, USA

    Sophie Pautot, Hanson Lee & Ehud Y Isacoff

  3. Department of Chemistry, University of California Berkeley, Berkeley, 94720, California, USA

    Jay T Groves

Authors
  1. Sophie Pautot
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hanson Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ehud Y Isacoff
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jay T Groves
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Ehud Y Isacoff or Jay T Groves.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pautot, S., Lee, H., Isacoff, E. et al. Neuronal synapse interaction reconstituted between live cells and supported lipid bilayers. Nat Chem Biol 1, 283–289 (2005). https://doi.org/10.1038/nchembio737

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nchembio737

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing