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NGL family PSD-95–interacting adhesion molecules regulate excitatory synapse formation

Nature Neuroscience volume 9, pages 1294–1301 (2006)Cite this article

Abstract

Synaptic cell adhesion molecules (CAMs) regulate synapse formation through their trans-synaptic and heterophilic adhesion. Here we show that postsynaptic netrin-G ligand (NGL) CAMs associate with netrin-G CAMs in an isoform-specific manner and, through their cytosolic tail, with the abundant postsynaptic scaffold postsynaptic density–95 (PSD-95). Overexpression of NGL-2 in cultured rat neurons increased the number of PSD-95–positive dendritic protrusions. NGL-2 located on heterologous cells or beads induced functional presynaptic differentiation in contacting neurites. Direct aggregation of NGL-2 on the surface membrane of dendrites induced the clustering of excitatory postsynaptic proteins. Competitive inhibition by soluble NGL-2 reduced the number of excitatory synapses. NGL-2 knockdown reduced excitatory, but not inhibitory, synapse numbers and currents. These results suggest that NGL regulates the formation of excitatory synapses.

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Figure 1: NGL interacts with PSD-95 through its C terminus and with netrin-G CAMs through its ectodomain in an isoform-specific manner.
Figure 2: NGL localizes to the postsynaptic side of excitatory synapses.
Figure 3: Overexpression of NGL-2 increases the number of PSD-95–positive dendritic protrusions through both PSD-95–dependent and PSD-95–independent mechanisms.
Figure 4: NGL-2 expression in heterologous cells induces functional presynaptic differentiation in contacting neurites.
Figure 5: Beads bearing the ectodomain of NGL-2 induce functional excitatory presynaptic differentiation in contacting neurites.
Figure 6: Direct aggregation of NGL-2 on the dendritic surface induces clustering of excitatory postsynaptic proteins.
Figure 7: Soluble NGL-2 fusion proteins reduce the number of excitatory synapses.
Figure 8: Knockdown of NGL-2 by siRNA leads to morphological and functional loss of excitatory, but not inhibitory, synapses.

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Acknowledgements

This work was supported by the National Creative Research Initiative Program of the Korean Ministry of Science and Technology (E.K.), and by grants from the Korea Science and Engineering Foundation (M10500000049-05J0000-04900 to H.K.) and the US National Institutes of Health (NS-39444 to R.J.W.).

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Authors and Affiliations

  1. National Creative Research Initiative Center for Synaptogenesis and Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Korea

    Seho Kim, Seok-Kyu Kwon, Jooyeon Woo, Hyun Woo Lee, Karam Kim & Eunjoon Kim

  2. Department of Cell and Developmental Biology, University of North Carolina, Chapel Hill, 27599, North Carolina, USA

    Alain Burette & Richard J Weinberg

  3. Department of Anatomy and Division of Brain Korea 21, Biomedical Science, College of Medicine, Korea University, Seoul, 136-705, Korea

    Hye Sun Chung & Hyun Kim

  4. Neuroscience Center, University of North Carolina, Chapel Hill, 27599, North Carolina, USA

    Richard J Weinberg

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Contributions

A.B. and R.J.W. performed the light and electron microscopic studies. H.S.C. and H.K. conducted the in situ hybridization analysis. S.-K.K. performed the in vivo coimmunoprecipitation. J.W. did the coclustering assay. H.W.L. and K.K. performed the northern blot analysis.

Note: Supplementary information is available on the Nature Neuroscience website.

Corresponding author

Correspondence to Eunjoon Kim.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Amino acid sequence alignment of NGL family proteins. (PDF 1845 kb)

Supplementary Fig. 2

Interaction between NGL and PSD-95. (PDF 727 kb)

Supplementary Fig. 3

NGL and netrin-G mediates cell adhesion in an isoform-specific manner. (PDF 773 kb)

Supplementary Fig. 4

Expression patterns of NGL mRNAs and proteins. (PDF 1651 kb)

Supplementary Fig. 5

Synaptic localization of NGL proteins in brain. (PDF 727 kb)

Supplementary Fig. 6

Polarized distribution of netrin-G2 to axons in cultured neurons. (PDF 577 kb)

Supplementary Fig. 7

Characterization of NGL-2 siRNA. (PDF 679 kb)

Supplementary Methods (PDF 60 kb)

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Kim, S., Burette, A., Chung, H. et al. NGL family PSD-95–interacting adhesion molecules regulate excitatory synapse formation. Nat Neurosci 9, 1294–1301 (2006). https://doi.org/10.1038/nn1763

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