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GDNF and GFRα1 promote formation of neuronal synapses by ligand-induced cell adhesion

Nature Neuroscience volume 10, pages 293–300 (2007)Cite this article

Abstract

The establishment of synaptic connections requires precise alignment of pre- and postsynaptic terminals. The glial cell line–derived neurotrophic factor (GDNF) receptor GFRα1 is enriched at pre- and postsynaptic compartments in hippocampal neurons, suggesting that it has a function in synapse formation. GDNF triggered trans-homophilic binding between GFRα1 molecules and cell adhesion between GFRα1-expressing cells. This represents the first example of a cell-cell interaction being mediated by a ligand-induced cell adhesion molecule (LICAM). In the presence of GDNF, ectopic GFRα1 induced localized presynaptic differentiation in hippocampal neurons, as visualized by clustering of vesicular proteins and neurotransmitter transporters, and by activity-dependent vesicle recycling. Presynaptic differentiation induced by GDNF was markedly reduced in neurons lacking GFRα1. Gdnf mutant mice showed reduced synaptic localization of presynaptic proteins and a marked decrease in the density of presynaptic puncta, indicating a role for GDNF signaling in hippocampal synaptogenesis in vivo. We propose that GFRα1 functions as a LICAM to establish precise synaptic contacts and induce presynaptic differentiation.

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Figure 1: Developmental expression and synaptic localization of GDNF receptors in hippocampal neurons.
Figure 2: GDNF-dependent trans-homophilic interactions between GFRα1 molecules promote cell adhesion.
Figure 3: GDNF increases the colocalization of pre- and postsynaptic markers, but not the expression of presynaptic proteins, in hippocampal neurons.
Figure 4: Localized induction of presynaptic assembly by immobilized GFRα1 in the presence of GDNF.
Figure 5: Excitatory and inhibitory presynaptic differentiation, activity-dependent vesicle recycling, and requirement of GFRα1 and NCAM but not RET.
Figure 6: Requirement of GDNF signaling for hippocampal presynaptic assembly and maturation in vivo.

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Acknowledgements

We thank D. Sjöstrand for western blot of NCAM and GFRα1 in COS cells, A. Moliner for help with Jurkat cell cultures, R. Scott for deletion constructs of GFRα1, T. Harkany, E. Restrepo, B. Stevens and B. Barres for experimental advice and W. Friedman, M. Fainzilber, A. Canty and L. Reichardt for comments and suggestions. This work was funded by grants from the Swedish Foundation for Strategic Research, the Swedish Research Council (33X-10908-10A) and the European Commission (QLG3-CT-2002-01000). F.L. was partially supported by the David och Astrid Hageléns Foundation, the Swedish Research Council (33P-15416-01A) and the Foundation for Geriatric Disease and Karolinska Institute, and G.P. by the Swedish Research Council (33PS-14809-01A). T.S.-G. was partially supported by the European Union NeuroNE network (LSHM-CT-2004-512039).

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Authors and Affiliations

  1. Division of Molecular Neurobiology, Department of Neuroscience, Karolinska Institute, 17177, Stockholm, Sweden

    Fernanda Ledda, Gustavo Paratcha, Tatiana Sandoval-Guzmán & Carlos F Ibáñez

  2. Department of Neuroscience, Laboratory of Molecular and Cellular Neuroscience, Karolinska Institute, 17177, Stockholm, Sweden

    Fernanda Ledda & Gustavo Paratcha

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Supplementary information

Supplementary Fig. 1

Expression of GDNF receptors in hippocampus. (PDF 114 kb)

Supplementary Fig. 2

Comparison between the adhesive activities of GFRα1 and NCAM. (PDF 611 kb)

Supplementary Fig. 3

Lack of heterophilic interactions in trans between GFRα1 and NCAM. (PDF 685 kb)

Supplementary Fig. 4

Trans-homophilic interactions between GFRα1 molecules require an intact GDNF binding domain. (PDF 615 kb)

Supplementary Fig. 5

GDNF regulates the aggregation, but not the expression, of vesicle-associated pre-synaptic proteins in hippocampal neurons. (PDF 701 kb)

Supplementary Fig. 6

Localized induction of presynaptic differentiation by immobilized GFRα proteins in the presence of GDNF ligands. (PDF 697 kb)

Supplementary Fig. 7

GDNF binding to NCAM in hippocampal synaptosomes and Gdnf mRNA expression in P15 Gdnf+/− hippocampus. (PDF 84 kb)

Supplementary Fig. 8

Synaptophysin content in synapses of P15 and 6-week-old Gdnf+/− mice. (PDF 583 kb)

Supplementary Fig. 9

Schematic of synapse formation by ligand-induced cell adhesion. (PDF 524 kb)

Supplementary Fig. 10

Controls for Jurkat cell adhesion experiments. (PDF 610 kb)

Supplementary Methods (PDF 127 kb)

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Ledda, F., Paratcha, G., Sandoval-Guzmán, T. et al. GDNF and GFRα1 promote formation of neuronal synapses by ligand-induced cell adhesion. Nat Neurosci 10, 293–300 (2007). https://doi.org/10.1038/nn1855

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