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:

Glutamate receptors regulate actin-based plasticity in dendritic spines

Nature Neuroscience volume 3, pages 887–894 (2000)Cite this article

Abstract

Dendritic spines at excitatory synapses undergo rapid, actin-dependent shape changes which may contribute to plasticity in brain circuits. Here we show that actin dynamics in spines are potently inhibited by activation of either AMPA or NMDA subtype glutamate receptors. Activation of either receptor type inhibited actin-based protrusive activity from the spine head. This blockade of motility caused spines to round up so that spine morphology became both more stable and more regular. Inhibition of spine motility by AMPA receptors was dependent on postsynaptic membrane depolarization and influx of Ca2+ through voltage-activated channels. In combination with previous studies, our results suggest a two-step process in which spines initially formed in response to NMDA receptor activation are subsequently stabilized by AMPA receptors.

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: Glutamate inhibits actin dynamics in dendritic spines.
Figure 2: AMPA receptor stimulation inhibits actin-based dendritic spine motility.
Figure 3: AMPA inhibits spine motility in organotypic slice cultures from hippocampus.
Figure 4: Inhibition of spine motility by NMDA requires lowering of the external Mg2+ concentration.
Figure 5: AMPA receptor activation leads to spine rounding.
Figure 6: Inhibition of spine motility by AMPA depends on extracellular Na+.
Figure 7: Membrane depolarization inhibits spine motility.

Similar content being viewed by others

References

  1. Feldman, D. E. & Knudsen, E. I. Experience-dependent plasticity and the maturation of glutamatergic synapses. Neuron 20, 1067–1071 (1998).

    CAS  PubMed  Google Scholar 

  2. Gray, E. G. Electron microscopy of synaptic contacts on dendritic spines of the cerebral cortex. Nature 183, 1592– 1593 (1959).

    CAS  PubMed  Google Scholar 

  3. Carlin, R. K. & Siekevitz, P. Plasticity in the central nervous system: do synapses divide? Proc. Natl. Acad. Sci. USA 80, 3517–3521 (1983).

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Harris, K. M. & Kater, S. B. Dendritic spines: cellular specializations imparting both stability and flexibility to synaptic function. Annu. Rev. Neurosci. 17, 341–371 (1994).

    CAS  PubMed  Google Scholar 

  5. Frotscher, M., Mannsfeld, B. & Wenzel, J. Environmentally affected differentiation of dendrite spines of pyramidal neurons in rat hippocampus (CA1). J. Hirnforsch 16, 443–450 (1975).

    CAS  PubMed  Google Scholar 

  6. Moser, M. B., Trommald, M. & Andersen, P. An increase in dendritic spine density on hippocampal CA1 pyramidal cells following spatial learning in adult rats suggests the formation of new synapses. Proc. Natl. Acad. Sci. USA 91, 12673–12675 (1994).

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Muller, D. Ultrastructural plasticity of excitatory synapses. Rev. Neurosci. 8, 77–93 (1997 ).

    CAS  PubMed  Google Scholar 

  8. Dailey, M. E. & Smith, S. J. The dynamics of dendritic structure in developing hippocampal slices. J. Neurosci. 16, 2983–2994 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Ziv, N. E. & Smith, S. J. Evidence for a role of dendritic filopodia in synaptogenesis and spine formation. Neuron 17, 91–102 (1996).

    CAS  PubMed  Google Scholar 

  10. Fischer, M., Kaech, S., Knutti, D. & Matus, A. Rapid actin-based plasticity in dendritic spines. Neuron 20, 847–854 (1998).

    CAS  PubMed  Google Scholar 

  11. Maletic-Savatic, M., Malinow, R. & Svoboda, K. Rapid dendritic morphogenesis in CA1 hippocampal dendrites induced by synaptic activity. Science 283, 1923–1927 (1999).

    CAS  PubMed  Google Scholar 

  12. Engert, F. & Bonhoeffer, T. Dendritic spine changes associated with hippocampal long-term synaptic plasticity. Nature 399, 66–70 (1999).

    CAS  PubMed  Google Scholar 

  13. Dunaevsky, A., Tashiro, A., Majewska, A., Mason, C. & Yuste, R. Developmental regulation of spine motility in the mammalian central nervous system. Proc. Natl. Acad. Sci. USA 96, 13438–13443 (1999).

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Lendvai, B., Stern, E. A., Chen, B. & Svoboda, K. Experience-dependent plasticity of dendritic spines in the developing rat barrel cortex in vivo. Nature 404, 876–881 (2000).

    CAS  PubMed  Google Scholar 

  15. McKinney, R. A., Capogna, M., Durr, R., Gahwiler, B. H. & Thompson, S. M. Miniature synaptic events maintain dendritic spines via AMPA receptor activation. Nat. Neurosci. 2, 44–49 (1999).

    CAS  PubMed  Google Scholar 

  16. Toni, N., Buchs, P. A., Nikonenko, I., Bron, C. R. & Muller, D. LTP promotes formation of multiple spine synapses between a single axon terminal and a dendrite. Nature 402, 421–425 (1999).

    CAS  PubMed  Google Scholar 

  17. Kaech, S., Fisher, M., Doll, T. & Matus, A. Isoform specificity in the relationship of actin to dendritic spines. J. Neurosci. 17, 9565–9572 (1997).

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Marin-Padilla, M. Number and distribution of apical dendritic spines of the layer V pyramidal cells in man. J. Comp. Neurol. 131, 475– 490 (1967).

    CAS  PubMed  Google Scholar 

  19. Spacek, J. & Hartmann, M. Three-dimensional analysis of dendritic spines. I. Quantitative observations related to dendritic spine and synaptic morphology in cerebral and cerebellar cortices. Anat. Embryol. (Berl.) 167, 289–310 (1983).

    CAS  Google Scholar 

  20. Choi, D. W. Glutamate receptors and the induction of excitotoxic neuronal death. Prog. Brain. Res. 100, 47–51 (1994).

    CAS  PubMed  Google Scholar 

  21. Halpain, S., Hipolito, A. & Saffer, L. Regulation of F-actin stability in dendritic spines by glutamate receptors and calcineurin. J. Neurosci. 18, 9835–9844 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  22. 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).

    CAS  PubMed  Google Scholar 

  23. Nowak, L., Bregestovski, P., Ascher, P., Herbet, A. & Prochiantz, A. Magnesium gates glutamate-activated channels in mouse central neurones. Nature 307, 462–465 (1984).

    CAS  PubMed  Google Scholar 

  24. Allison, D. W., Gelfand, V. I., Spector, I. & Craig, A. M. Role of actin in anchoring postsynaptic receptors in cultured hippocampal neurons: differential attachment of NMDA versus AMPA receptors. J. Neurosci. 18, 2423–2436 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Guthrie, P. B., Segal, M. & Kater, S. B. Independent regulation of calcium revealed by imaging dendritic spines. Nature 354, 76– 80 (1991).

    CAS  PubMed  Google Scholar 

  26. Denk, W., Sugimori, M. & Llinas, R. Two types of calcium response limited to single spines in cerebellar Purkinje cells. Proc. Natl. Acad. Sci. USA 92, 8279–8282 (1995).

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Koester, H. J. & Sakmann, B. Calcium dynamics in single spines during coincident pre- and postsynaptic activity depend on relative timing of back-propagating action potentials and subthreshold excitatory postsynaptic potentials. Proc. Natl. Acad. Sci. USA 95, 9596–9601 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Furukawa, K. et al. The actin-severing protein gelsolin modulates calcium channel and NMDA receptor activities and vulnerability to excitotoxicity in hippocampal neurons. J. Neurosci. 17, 8178– 8186 (1997).

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Matus, A. Postsynaptic actin and neuronal plasticity. Curr. Opin. Neurobiol. 9, 561–565 (1999).

    CAS  PubMed  Google Scholar 

  30. Hollmann, M., Hartley, M. & Heinemann, S. Ca2+ permeability of KA-AMPA-gated glutamate receptor channels depends on subunit composition. Science 252, 851–853 (1991).

    CAS  PubMed  Google Scholar 

  31. Geiger, J. R. et al. Relative abundance of subunit mRNAs determines gating and Ca2+ permeability of AMPA receptors in principal neurons and interneurons in rat CNS. Neuron 15, 193– 204 (1995).

    CAS  PubMed  Google Scholar 

  32. Markram, H. & Sakmann, B. Calcium transients in dendrites of neocortical neurons evoked by single subthreshold excitatory postsynaptic potentials via low-voltage-activated calcium channels. Proc. Natl. Acad. Sci. USA 91, 5207–5211 (1994).

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Avery, R. B. & Johnston, D. Multiple channel types contribute to the low-voltage-activated calcium current in hippocampal CA3 pyramidal neurons. J. Neurosci. 16, 5567– 5582 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  34. McDonough, S. I. & Bean, B. P. Mibefradil inhibition of T-type calcium channels in cerebellar purkinje neurons. Mol. Pharmacol. 54, 1080–1087 (1998).

    CAS  PubMed  Google Scholar 

  35. Bartlett, W. P. & Banker, G. A. An electron microscopic study of the development of axons and dendrites by hippocampal neurons in culture. II. Synaptic relationships. J. Neurosci. 4, 1954–1965 (1984).

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Boyer, C., Schikorski, T. & Stevens, C. F. Comparison of hippocampal dendritic spines in culture and in brain. J. Neurosci. 18, 5294– 5300 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Gahwiler, B. H. Development of the hippocampus in vitro: cell types, synapses and receptors. Neuroscience 11, 751– 760 (1984).

    CAS  PubMed  Google Scholar 

  38. Magee, J., Hoffman, D., Colbert, C. & Johnston, D. Electrical and calcium signaling in dendrites of hippocampal pyramidal neurons. Annu. Rev. Physiol. 60, 327–346 (1998).

    CAS  PubMed  Google Scholar 

  39. Svoboda, K. & Mainen, Z. F. Synaptic [Ca2+]: intracellular stores spill their guts. Neuron 22, 427–430 (1999).

    CAS  PubMed  Google Scholar 

  40. Liao, D., Zhang, X., O'Brien, R., Ehlers, M. D. & Huganir, R. L. Regulation of morphological postsynaptic silent synapses in developing hippocampal neurons. Nat. Neurosci. 2, 37–43 (1999 ).

    CAS  PubMed  Google Scholar 

  41. Petralia, R. S. et al. Selective acquisition of AMPA receptors over postnatal development suggests a molecular basis for silent synapses. Nat. Neurosci. 2, 31–36 (1999 ).

    CAS  PubMed  Google Scholar 

  42. Shi, S. H. et al. Rapid spine delivery and redistribution of AMPA receptors after synaptic NMDA receptor activation. Science 284 , 1811–1816 (1999).

    CAS  PubMed  Google Scholar 

  43. Kater, S. B. & Mills, L. R. Regulation of growth cone behavior by calcium. J. Neurosci. 11, 891– 899 (1991).

    CAS  PubMed  PubMed Central  Google Scholar 

  44. Hong, K., Nishiyama, M., Henley, J., Tessier-Lavigne, M. & Poo, M. Calcium signalling in the guidance of nerve growth by netrin-1. Nature 403, 93 –98 (2000).

    CAS  PubMed  Google Scholar 

  45. Zheng, J. Q. Turning of nerve growth cones induced by localized increases in intracellular calcium ions. Nature 403, 89– 93 (2000).

    CAS  PubMed  Google Scholar 

  46. Banker, G. & Goslin, K. in Culturing Nerve Cells (eds. Banker, G. & Goslin, K.) 251–281 (Bradford Books, MIT Press, Cambridge, 1991).

    Google Scholar 

  47. Gahwiler, B. H., Thompson, S. M., Audinat, E. & Robertson, R. T. in Culturing Nerve Cells (eds. Banker, G. & Goslin, K.) 379–411 (Bradford Books, MIT Press, Cambridge, Massachusetts, 1991).

    Google Scholar 

Download references

Acknowledgements

We thank Beat Ludin for assistance with image analysis and John Kemp for supplying mibefradil.

Author information

Author notes

  1. Maria Fischer and Stefanie Kaech: The first two authors contributed equally to this work

Authors and Affiliations

  1. Friedrich Miescher Institute, P.O. Box 2543, Basel, 4002, Switzerland

    Maria Fischer, Stefanie Kaech, Uta Wagner, Heike Brinkhaus & Andrew Matus

Authors
  1. Maria Fischer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Stefanie Kaech
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Uta Wagner
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Heike Brinkhaus
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Andrew Matus
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Andrew Matus.

Supplementary information

Figure 1

Glutamate blocks actin dynamics in dendritic spines. This Time Movie-lapse sequence shows GFP-actin dynamics in part of the dendrite shown in Fig. 1. The first section was recorded under control conditions followed by a sequence recorded after 100 µM glutamate had been added to the medium together with 100 µM APV. The high level of actin dynamics seen in these spines under control conditions is typical of all cells maintained in low density culture.

QuickTime Movie Figure 1 (MOV 2.4 MB)

Figure 2

AMPA blocks spine motility in the presence of NMDA receptor antagonists. Effect of AMPA in the presence of MK801. There are two versions of this sequence, Fig2a_1.mov showing the original low magnification recording of a long stretch of dendrite and Fig2a_2.mov showing a small extract of the same sequence at higher magnification. The effects of AMPA both alone and in the presence of the NMDA receptor channel blocker MK801 (10 µM) are documented in this single recording. The first half of this sequence, showing the reversible effect of AMPA on a long stretch of dendrite, was used to prepare the difference images shown in Fig. 5. The sequence shows: 1. (no label) the control condition, when spines are motile; 2. (labeled "+AMPA") a brief period in which the medium was switched to one containing 2 µM AMPA, when spine motility was blocked. Note the rounding up of the spine heads; 3. (no label) an intermediate period when the AMPA was washed out to demonstrate that spine motility recovers; 4. (labeled "+MK801") after the AMPA receptor antagonist MK801 was added to the medium, when spine motility continued unaffected; 5. (labeled +AMPA + MK801) after AMPA was added to the medium in addition to the MK801. During this long final period spine motility was inhibited, showing that it is produced by AMPA receptor stimulation independently of the blocked NMDA receptors.

Figure 2b: Effect of AMPA in the presence of APV. This sequence focuses on part of a dendrite bearing spines with a variety of characteristic morphologies including long necked, mushroom shaped and stubby. The two extracts are from a single long experiment, the first recorded under control conditions, the second in the presence of 1 µM AMPA. 100 µM APV was present throughout the recording to block NMDA receptors. Spines of all categories show actin-based motility under control conditions which is blocked by AMPA. Note that, as in the example shown in Fig2a_2.mov, the effect of AMPA receptor activation is to inhibit the formation of actin-rich protrusions from the spine head.

QuickTime Movie Figure 2a_1 (MOV 3.2 MB)

QuickTime Movie Figure 2a_2 (MOV 4.3 MB)

QuickTime Movie Figure 2b (MOV 2.4 MB)

Figure 3

AMPA blocks spine actin dynamics in organotypic slice cultures. Three video sequences, taken from a time-lapse recording of the 4 week-old slice culture shown in the still frames in the top row of Fig. 3. The sequences are labeled as follows: Fig3_1.mov, spine dynamics in the control condition; Fig3_2.mov recorded in the presence of 1µM AMPA; Fig3_3.mov after washout of the AMPA. Each sequence represents 15 min of recording comprising 60 frames taken at 15 sec intervals. Note the lack of spine dynamics during the sequence shown in Fig3_2, indicating that spine movement seen under control conditions is due to actin dynamics and not focus shift or other artifacts of the recording technique.

QuickTime Movie Figure 3_1 (MOV 2.4 MB)

QuickTime Movie Figure 3_2 (MOV 2.6 MB)

QuickTime Movie Figure 3_3 (MOV 2.5 MB)

Figure 4

Blockade of spine motility by NMDA requires lowering Mg2+ in the medium. This recording begins with a period of control recording followed by a sequence in which 1 µM NMDA was added in regular medium containing 0.5 mM Mg2+ (labeled "+NMDA") which did not inhibit spine motility. The medium was then switched to one containing NMDA and 0.1 mM Mg2+ (labeled +NMDA + low Mg2+) when spine motility was inhibited.

QuickTime Movie Figure 4 (MOV 4.1 MB)

Figure 6

AMPA receptor blockade of spine motility is Na+ dependent. A single time-lapse recording during which the following conditions were applied: 1. (labeled "Control") in control medium the spines on this segment of dendrite were motile; 2. (labeled "-Na") when NaCl was replaced by choline chloride spine motility continued undiminished; 3. (labeled "-Na, +AMPA") subsequently 200nM AMPA was added to the medium but still in the absence of sodium. Despite the presence of AMPA, motility was not noticeably inhibited when Na+ was absent; 4. (labeled "+Na+AMPA") When NaCl was added back to the medium AMPA now blocked spine motility.

Additional data. Blockade of voltage dependent Na+ channels does not affect AMPA-induced inhibition of spine motility. The first part of this recording shows that spine motility remains intact in a cell that had been exposed to 1µM TTX for 2 hours. The second part of the recording shows that adding AMPA (2 µM) still inhibited spine motility even though TTX was present. This particular cell had a well-developed set of recurrent axons which are visible among its dendrites as thinner processes containing faint patches of actin. These are seen to travel along the axons during the recording and this transport is not blocked by AMPA treatment.

QuickTime Movie Figure 6 (MOV 2.4 MB)

QuickTime Movie Figure 6 Additional Data (MOV 26.1 MB)

Figure 7

Raising [K+]o induces transitory blockade of spine motility. At the time indicated by the appearance of the label "KCl" the external K+ concentration was raised to 8 mM. This produced a transitory inhibition of spine motility and rounding up of spine heads, consistent with the spontaneous recovery of the membrane potential from the temporary depolarization induced by raising the external K+ concentration.

Supplementary data_cd. Voltage-activated calcium channel (VAC) antagonists and AMPA-induced inhibition of spine motility. Fig7_Ni: These 4 excerpts are taken from a single continuous time-lapse recording under the following conditions: 1) a period of control recording in regular Tyrode's solution to establish the base level of motility. 2) After the addition of 100 µM Ni2+, which does not influence spine motility. 3) With the addition of 2 µM AMPA in the continued presence of 10 µM Ni2+. The usual inhibition of motility by AMPA is suppressed by the Ni2+. 4) After changing the medium to one still containing AMPA but now lacking Ni2+. Spine motility is now shows AMPA inhibition. APV (100 µM) to block NMDA receptors was present in all solutions.

Supplementary data_ni. The equivalent experiment to the one described above showing that AMPA can inhibit spine motility in the presence of 500 µM Cd2+.

Supplementary data_nif. Showing that the HVAC antagonist nifedipine (20 µM) does not suppress AMPA inhibition of spine motility.

QuickTime Movie Figure 7 (MOV 3.7 MB)

QuickTime Movie Figure 7 Supplementary Data cd (MOV 2.4 MB)

QuickTime Movie Figure 7 Supplementary Data ni (MOV 224 KB)

QuickTime Movie Figure 7 Supplementary Data nif (MOV 2.4 MB)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fischer, M., Kaech, S., Wagner, U. et al. Glutamate receptors regulate actin-based plasticity in dendritic spines. Nat Neurosci 3, 887–894 (2000). https://doi.org/10.1038/78791

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/78791

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