Abstract
Many excitatory synapses are thought to be postsynaptically 'silent', possessing functional NMDA but lacking functional AMPA glutamate receptors. The acquisition of AMPA receptors at silent synapses may be important in synaptic plasticity and neuronal development. Here we characterize a possible morphological correlate of silent synapses in cultured hippocampal neurons. Initially, most excitatory synapses contained NMDA receptors, but only a few contained detectable AMPA receptors. Synapses progressively acquired AMPA receptors as the cultures matured. AMPA receptor blockade increased the number, size and fluorescent intensity of AMPA receptor clusters and rapidly induced the appearance of AMPA receptors at 'silent' synapses. In contrast, NMDA receptor blockade increased the size, intensity and number of NMDA receptor clusters and decreased the number of AMPA receptor clusters, resulting in an increase in the proportion of 'silent' synapses. These results suggest that the number of silent synapses is regulated during development and by changes in synaptic activity.
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References
Feldman, D. E. & Knudsen, E. I. Experience–dependent plasticity and the maturation of glutamatergic synapses. Neuron 20, 1067–1071 (1998).
Malenka, R. C. & Nicoll, R. A. Silent synapses speak up. Neuron 19, 473–476 ( 1997).
Hollmann, M. & Heinemann, S. Cloned glutamate receptors. Annu. Rev. Neurosci. 17, 31–108 (1994).
Liao, D., Hessler, N. A. & Malinow, R. Activation of postsynaptically silent synapses during pairing–induced LTP in CA1 region of hippocampal slice. Nature 375, 400–404 ( 1995).
Isaac, J. T., Nicoll, R. A. & Malenka, R. C. Evidence for silent synapses: implications for the expression of LTP. Neuron 15, 427– 434 (1995).
Durand, G., Kovalchuk, Y. & Konnerth, A. Long–term potentiation and functional synapse induction in developing hippocampus. Nature 381, 71–75 (1996).
Wu, G., Malinow, R. & Cline, H. T. Maturation of a central glutamatergic synapse. Science 274, 972–976 ( 1996).
Isaac, J. T., Crair, M. C., Nicoll, R. A. & Malenka, R. C. Silent synapses during development of thalamocortical inputs. Neuron 18, 269–280 ( 1997).
Nowak, L., Bregestovski, P., Ascher, P., Herbet, A. & Prochiantz, A. Magnesium gates glutamate–activated channels in mouse central neurones. Nature 307, 462–465 (1984).
Mayer, M. L., Westbrook, G. L. & Guthrie, P. B. Voltage–dependent block by Mg2+ of NMDA responses in spinal cord neurones. Nature 309 , 261–263 (1984).
Kullmann, D. M. & Asztely, F. Extrasynaptic glutamate spillover in the hippocampus: evidence and implications. Trends Neurosci. 21, 8–14 (1998).
Asztely, F., Erdemli, G. & Kullmann, D. M. Extrasynaptic glutamate spillover in the hippocampus: dependence on temperature and the role of active glutamate uptake. Neuron 18, 281–293 ( 1997).
O'Brien, R. J. et al. The development of excitatory synapses in cultured spinal neurons. J. Neurosci. 17, 7339– 7350 (1997).
Mammen, A. L., Huganir, R. L. & O'Brien, R. J. Redistribution and stabilization of cell surface glutamate receptors during synapse formation. J. Neurosci. 17, 7351–7358 (1997).
Kim, J. H., Liao, D., Lau, L. F. & Huganir, R. L. SynGAP: a synaptic RasGAP that associates with the PSD–95/SAP90 protein family. Neuron 20, 683–691 ( 1998).
Kirsch, J. & Betz, H. Glycine–receptor activation is required for receptor clustering in spinal neurons. Nature 392, 717–720 (1998).
Kirsch, J., Wolters, I., Triller, A. & Betz, H. Gephyrin antisense oligonucleotides prevent glycine receptor clustering in spinal neurons. Nature 366, 745–748 ( 1993).
Rao, A., Kim, E., Sheng, M. & Craig, A. M. Heterogeneity in the molecular composition of excitatory postsynaptic sites during development of hippocampal neurons in culture. J. Neurosci. 18, 1217–1229 (1998).
Rao, A. & Craig, M. Activity regulates the synaptic localization of the NMDA receptor in hippocampal neurons. Neuron 19, 801–812 (1997).
Turrigiano, G. G., Leslie, K. R., Desai, N. S., Rutherford, L. C. & Nelson, S. B. Activity–dependent scaling of quantal amplitude in neocortical neurons. Nature 391, 892–896 (1998).
Baranes, D., Lopez–Garcia, J. C., Chen, M., Bailey, C. H. & Kandel, E. R. Reconstitution of the hippocampal mossy fiber and associational–commissural pathways in a novel dissociated cell culture system. Proc. Natl. Acad. Sci. USA 93, 4706–4711 (1996).
Amaral, D. G. & Dent, J. A. Development of the mossy fibers of the dentate gyrus: I. A light and electron microscopic study of the mossy fibers and their expansions. J. Comp. Neurol. 195, 51–86 (1981).
Claiborne, B. J., Amaral, D. G. & Cowan, W. M. A light and electron microscopic analysis of the mossy fibers of the rat dentate gyrus. J. Comp. Neurol. 246, 435–458 (1986).
Chicurel, M. E. & Harris, K. M. Three–dimensional analysis of the structure and composition of CA3 branched dendritic spines and their synaptic relationships with mossy fiber boutons in the rat hippocampus. J. Comp. Neurol. 325, 169– 182 (1992).
Harris, E. W. & Cotman, C. W. Long–term potentiation of guinea pig mossy fiber responses is not blocked by N–methyl–D–aspartate antagonists. Neurosci. Lett. 70, 132– 137 (1986).
Zalutsky, R. A. & Nicoll, R. A. Comparison of two forms of long–term potentiation in single hippocampal neurons [published erratum appears in Science 251, 856, 1991]. Science 248, 1619–1624 ( 1990).
Ben–Ari, Y., Khazipov, R., Leinekugel, X., Caillard, O. & Gaiarsa, J. L. GABAA, NMDA and AMPA receptors: a developmentally regulated 'menage a trois'. Trends Neurosci. 20, 523–529 ( 1997).
Nusser, Z. et al. Cell type and pathway dependence of synaptic AMPA receptor number and variability in the hippocampus. Neuron 21 , 545–559 (1998).
Lissin, D. V. et al. Activity differentially regulates the surface expression of synaptic AMPA and NMDA glutamate receptors. Proc. Natl. Acad. Sci. USA 95, 7097–7102 ( 1998).
O'Brien, R. J., Lau, L. F. & Huganir, R. L. Molecular mechanisms of glutamate receptor clustering at excitatory synapses. Curr. Opin. Neurobiol. 8, 364–369 (1998).
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).
Dong, H. et al. GRIP: a synaptic PDZ domain–containing protein that interacts with AMPA receptors. Nature 386, 279– 284 (1997).
Osten, P. et al. The AMPA receptor GluR2 C terminus can mediate a reversible, ATP–dependent interaction with NSF and alpha– and beta–SNAPs. Neuron 21, 99–110 (1998).
Nishimune, A. et al. NSF binding to GluR2 regulates synaptic transmission. Neuron 21, 87–97 ( 1998).
Song, I. et al. Interaction of the N–ethylmaleimide sensitive factor with AMPA receptors. Neuron 21, 393– 400 (1998).
Banker, G. A. & Cowan, W. M. Rat hippocampal neurons in dispersed cell culture. Brain Res. 126, 397– 425 (1977).
Goslin, K. & Banker, G. in Culturing Nerve Cells (eds Banker, G. & Goslin, K.) 251–283 (MIT Press, London, 1991).
Malgaroli, A. & Tsien, R. W. Glutamate–induced long–term potentiation of the frequency of miniature synaptic currents in cultured hippocampal neurons. Nature 357, 134– 139 (1992).
Acknowledgements
We thank C. Doherty and J. Bernhardt for technical support and D. Bury and J. Kim for assistance in preparing the manuscript. This work was supported by the Howard Hughes Medical Institute and the National Institutes of Health (R.L.H.).
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Liao, D., Zhang, X., O'Brien, R. et al. Regulation of morphological postsynaptic silent synapses in developing hippocampal neurons. Nat Neurosci 2, 37–43 (1999). https://doi.org/10.1038/4540
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DOI: https://doi.org/10.1038/4540
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