Abstract
The ability of nerve cells to receive up to several thousands of synaptic inputs from other neurons provides the anatomical basis for information processing in the vertebrate brain. The formation of functional synapses involves selective clustering of neurotransmitter receptors at presumptive postsynaptic regions of the neuronal plasma membrane1,2,3,4. Receptor-associated proteins are believed to be crucial for this process. In spinal neurons, synaptic targeting of the inhibitory glycine receptor (GlyR)5,6 depends on the expression of the anchoring protein gephyrin7,8,9. Here we show that the competitive GlyR antagonist strychnine and L-type Ca2+-channel blockers inhibit the accumulation of GlyR and gephyrin at postsynaptic membrane areas in cultured rat spinal neurons. Our data are consistent with a model in which GlyR activation that results in Ca2+ influx is required for the clustering of gephyrin and GlyR at developing postsynaptic sites. Similar activity-driven mechanisms may be of general importance in synaptogenesis.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Froehner, S. C. Regulation of ion channel distribution at synapses. Annu. Rev. Neurosci. 16, 347–368 (1993).
Ehlers, M. D., Mammen, A. L., Lau, L.-F. & Huganir, R. L. Synaptic targeting of glutamate receptors. Curr. Opin. Cell Biol. 8, 484–489 (1996).
Kirsch, J., Meyer, G. & Betz, H. Synaptic targeting of ionotropic neurotransmitter receptors. Mol. Cell. Neurosci. 8, 93–98 (1996).
Sheng, M. Glutamate receptors put in their place. Nature 386, 221–223 (1997).
Triller, A., Cluzeaud, F., Pfeiffer, F., Betz, H. & Korn, H. Distribution of glycine receptors at central synapses: an immunoelectron microscopy study. J. Cell Biol. 101, 683–688 (1985).
Altschuler, R. A., Betz, H., Parakkal, M., Reeks, K. & Wenthold, R. Identification of glycinergic synapses in the cochlear nucleus through immunocytochemical localization of the postsynaptic receptor. Brain Res. 369, 316–320 (1986).
Kirsch, J. et al. The 93-kDa glycine receptor-associated protein binds to tubulin. J. Biol. Chem. 266, 22242–22245 (1991).
Prior, P. et al. Primary structure and alternative splice variants of gephyrin, a putative glycine receptor-tubulin linker protein. Neuron 8, 1161–1170 (1992).
Kirsch, J., Wolters, I., Triller, A. & Betz, H. Gephyrin antisense oligonucleotides prevent glycine receptor clustering in spinal neurons. Nature 366, 745–748 (1993).
Kuhse, J., Betz, H. & Kirsch, J. The inhibitory glycine receptor: architecture, synaptic localization and molecular pathology of a postsynaptic ion channel complex. Curr. Opin. Neurobiol. 5, 318–323 (1995).
Hoch, W., Betz, H. & Becker, C.-M. Primary cultures of mouse spinal cord express the neonatal isoform of the inhibitory glycine receptor. Neuron 3, 339–348 (1990).
Béchade, C., Colin, I., Kirsch, J., Betz, H. & Triller, A. Glycine receptor α subunit and gephyrin expression in cultured spinal neurons: a quantitative analysis. Eur. J. Neurosci. 8, 429–435 (1996).
Bormann, J., Hamill, O. P. & Sakmann, B. Mechanism of anion permeation through channels gated by glycine and γ-aminobutyric acid in mouse cultured spinal neurones. J. Physiol. (London) 385, 243–286 (1987).
Kirsch, J. & Betz, H. The postsynaptic localization of the glycine receptor-associated protein gephyrin is regulated by the cytoskeleton. J. Neurosci. 15, 4148–4156 (1995).
Pfeiffer, F., Simler, R., Grenningloh, G. & Betz, H. Monoclonal antibodies and peptide mapping reveal structural similarities between the subunits of the glycine receptor of rat spinal cord. Proc. Natl Acad. Sci. USA 81, 7224–7227 (1984).
Knaus, P., Betz, H. & Rehm, H. Expression of synaptophysin during postnatal development of the mouse brain. J. Neurochem. 47, 1302–1304 (1986).
Virginio, C. & Cherubini, E. Glycine-activated whole cell and single channel currents in rat cerebellar granule cells in culture. Brain Res. 98, 30–40 (1997).
Fritschy, J. M. et al. Five subtypes of type A γ-aminobutyric acid receptors identified in neurons by double and triple immunofluorescence staining with subunit specific antibodies. Proc. Natl Acad. Sci. USA 89, 6726–6730 (1992).
Betz, H. The glycine receptor of rat spinal cord: exploring the site of action of the plant alkaloid strychnine. Angew. Chem. Int. Ed. 24, 365–370 (1985).
Becker, C.-M. in Handbook of Experimental Pharmacology, Vol. 102, Selective Neurotoxicity (eds Herken, H. & Hucho, F.) Vol. 1, 540–575 (Springer, Heidelberg, 1992).
Reichling, D. B., Kyrozis, A., Wang, J. & MacDermott, A. B. Mechanisms of GABA and glycine depolarization-induced calcium transients in rat dorsal horn neurons. J. Physiol. (London) 476, 411–421 (1994).
Wang, J., Reichling, D. B., Kyrozis, A. & MacDermott, A. B. Developmental loss of GABA- and glycine-induced depolarization and Ca2+ transients in embryonic rat dorsal horn neurons in culture. Eur. J.Neurosci. 6, 1275–1280 (1994).
Boehm, S., Harvey, R. J., v.Holst, A., Rohrer, H. & Betz, H. Glycine receptors in cultured chick sympathetic neurons are excitatory and trigger neurotransmitter release. J. Physiol. (London) 504, 683–694 (1997).
Craig, A. M., Banker, G., Chang, W., McGrath, M. E. & Serpinskaya, A. S. Clustering of gephyrin at GABAergic but not glutamatergic synapses in cultured rat hippocampal neurons. J. Neurosci. 16, 3166–3177 (1996).
Kirsch, J., Kuhse, J. & Betz, H. Targeting of glycine receptor subunits to gephyrin-rich domains in transfected human embryonic kidney cells. Mol. Cell. Neurosci. 6, 450–461 (1995).
Meyer, G., Kirsch, J., Betz, H. & Langosch, D. Identification of a gephyrin binding motif on the glycine receptor β subunit. Neuron 15, 563–572 (1995).
Hall, Z. W. & Sanes, J. R. Synaptic structure and development: the neuromuscular junction. Cell 72 (suppl.), 99–121 (1993).
Changeux, J. P. & Danchin, A. Selective stabilization of developing synapses as a mechanism for the specification of neuronal networks. Nature 264, 705–712 (1976).
Goslin, K. & Banker, G. in Culturing Nerve Cells (eds Banker, G. & Goslin, K.) 251–282 (MIT Press, Cambridge, MA, 1991).
Acknowledgements
We thank I. Bartnik for technical assistance and V. O'Connor, R. Harvey and A.Püschel for critically reading the manuscript. This work was supported by grants from the Deutsche Forschungsgemeinschaft and Fonds der Chemischen Industrie. J.K. holds an endowed professorship from the Stifterverband für die Deutsche Wissenschaft.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kirsch, J., Betz, H. Glycine-receptor activation is required for receptor clustering in spinal neurons. Nature 392, 717–720 (1998). https://doi.org/10.1038/33694
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/33694
This article is cited by
-
Induction of synapse formation by de novo neurotransmitter synthesis
Nature Communications (2022)
-
Enhancing neuronal chloride extrusion rescues α2/α3 GABAA-mediated analgesia in neuropathic pain
Nature Communications (2020)
-
Metabolomics methods to analyze full spectrum of amino acids in different domains of bovine colostrum and mature milk
European Food Research and Technology (2020)
-
Assembly and maintenance of GABAergic and Glycinergic circuits in the mammalian nervous system
Neural Development (2018)
-
Extrasynaptic homomeric glycine receptors in neurons of the rat trigeminal mesencephalic nucleus
Brain Structure and Function (2018)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.