Activity-dependent site-specific changes of glutamate receptor composition in vivo

Article metrics

Abstract

The subunit composition of postsynaptic non–NMDA-type glutamate receptors (GluRs) determines the function and trafficking of the receptor. Changes in GluR composition have been implicated in the homeostasis of neuronal excitability and synaptic plasticity underlying learning. Here, we imaged GluRs in vivo during the formation of new postsynaptic densities (PSDs) at Drosophila neuromuscular junctions coexpressing GluRIIA and GluRIIB subunits. GluR composition was independently regulated at directly neighboring PSDs on a submicron scale. Immature PSDs typically had large amounts of GluRIIA and small amounts of GluRIIB. During subsequent PSD maturation, however, the GluRIIA/GluRIIB composition changed and became more balanced. Reducing presynaptic glutamate release increased GluRIIA, but decreased GluRIIB incorporation. Moreover, the maturation of GluR composition correlated in a site-specific manner with the level of Bruchpilot, an active zone protein that is essential for mature glutamate release. Thus, we show that an activity-dependent, site-specific control of GluR composition can contribute to match pre- and postsynaptic assembly.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: GFP-labeled GluRIIA and GluRIIB expressed from genomic constructs are fully functional.
Figure 2: Patch-clamp recordings of NMJs expressing only GluRIIAGFP or GluRIIBGFP.
Figure 3: In vivo imaging of GluR composition during PSD maturation of developing Drosophila NMJs.
Figure 4: FRAP of GluRIIA and GluRIIB during synapse formation and maturation.
Figure 5: The BRP-dependent release component controls the maturation of GluR composition.
Figure 6: Influence of intracellular C-terminal domains on FRAP behavior in vivo.
Figure 7: Control of GluR composition and structural plasticity at the NMJ.

References

  1. 1

    McAllister, A.K. Dynamic aspects of CNS synapse formation. Annu. Rev. Neurosci. 30, 425–450 (2007).

  2. 2

    Garner, C.C., Zhai, R.G., Gundelfinger, E.D. & Ziv, N.E. Molecular mechanisms of CNS synaptogenesis. Trends Neurosci. 25, 243–251 (2002).

  3. 3

    Zhai, R.G. & Bellen, H.J. The architecture of the active zone in the presynaptic nerve terminal. Physiology (Bethesda) 19, 262–270 (2004).

  4. 4

    Witzemann, V. et al. Acetylcholine receptor epsilon–subunit deletion causes muscle weakness and atrophy in juvenile and adult mice. Proc. Natl. Acad. Sci. USA 93, 13286–13291 (1996).

  5. 5

    Takahashi, T. Postsynaptic receptor mechanisms underlying developmental speeding of synaptic transmission. Neurosci. Res. 53, 229–240 (2005).

  6. 6

    Derkach, V.A., Oh, M.C., Guire, E.S. & Soderling, T.R. Regulatory mechanisms of AMPA receptors in synaptic plasticity. Nat. Rev. Neurosci. 8, 101–113 (2007).

  7. 7

    Turrigiano, G. Homeostatic signaling: the positive side of negative feedback. Curr. Opin. Neurobiol. 17, 318–324 (2007).

  8. 8

    Schuster, C.M. et al. Molecular cloning of an invertebrate glutamate receptor subunit expressed in Drosophila muscle. Science 254, 112–114 (1991).

  9. 9

    Petersen, S.A., Fetter, R.D., Noordermeer, J.N., Goodman, C.S. & DiAntonio, A. Genetic analysis of glutamate receptors in Drosophila reveals a retrograde signal regulating presynaptic transmitter release. Neuron 19, 1237–1248 (1997).

  10. 10

    DiAntonio, A. Glutamate receptors at the Drosophila neuromuscular junction. Int. Rev. Neurobiol. 75, 165–179 (2006).

  11. 11

    Qin, G. et al. Four different subunits are essential for expressing the synaptic glutamate receptor at neuromuscular junctions of Drosophila. J. Neurosci. 25, 3209–3218 (2005).

  12. 12

    Featherstone, D.E. et al. An essential Drosophila glutamate receptor subunit that functions in both central neuropil and neuromuscular junction. J. Neurosci. 25, 3199–3208 (2005).

  13. 13

    Marrus, S.B., Portman, S.L., Allen, M.J., Moffat, K.G. & DiAntonio, A. Differential localization of glutamate receptor subunits at the Drosophila neuromuscular junction. J. Neurosci. 24, 1406–1415 (2004).

  14. 14

    DiAntonio, A., Petersen, S.A., Heckmann, M. & Goodman, C.S. Glutamate receptor expression regulates quantal size and quantal content at the Drosophila neuromuscular junction. J. Neurosci. 19, 3023–3032 (1999).

  15. 15

    Pawlu, C., DiAntonio, A. & Heckmann, M. Postfusional control of quantal current shape. Neuron 42, 607–618 (2004).

  16. 16

    Sigrist, S.J., Thiel, P.R., Reiff, D.F. & Schuster, C.M. The postsynaptic glutamate receptor subunit DGluR-IIA mediates long-term plasticity in Drosophila. J. Neurosci. 22, 7362–7372 (2002).

  17. 17

    Sigrist, S.J. et al. Postsynaptic translation affects the efficacy and morphology of neuromuscular junctions. Nature 405, 1062–1065 (2000).

  18. 18

    Rasse, T.M. et al. Glutamate receptor dynamics organizing synapse formation in vivo. Nat. Neurosci. 8, 898–905 (2005).

  19. 19

    Davis, G.W., DiAntonio, A., Petersen, S.A. & Goodman, C.S. Postsynaptic PKA controls quantal size and reveals a retrograde signal that regulates presynaptic transmitter release in Drosophila. Neuron 20, 305–315 (1998).

  20. 20

    Yoshihara, M., Adolfsen, B., Galle, K.T. & Littleton, J.T. Retrograde signaling by Syt 4 induces presynaptic release and synapse-specific growth. Science 310, 858–863 (2005).

  21. 21

    Reiff, D.F., Thiel, P.R. & Schuster, C.M. Differential regulation of active zone density during long-term strengthening of Drosophila neuromuscular junctions. J. Neurosci. 22, 9399–9409 (2002).

  22. 22

    Broadie, K.S. & Bate, M. Development of the embryonic neuromuscular synapse of Drosophila melanogaster. J. Neurosci. 13, 144–166 (1993).

  23. 23

    Nishikawa, K. & Kidokoro, Y. Junctional and extrajunctional glutamate receptor channels in Drosophila embryos and larvae. J. Neurosci. 15, 7905–7915 (1995).

  24. 24

    Campbell, R.E. et al. A monomeric red fluorescent protein. Proc. Natl. Acad. Sci. USA 99, 7877–7882 (2002).

  25. 25

    Sweeney, S.T., Broadie, K., Keane, J., Niemann, H. & O'Kane, C.J. Targeted expression of tetanus toxin light chain in Drosophila specifically eliminates synaptic transmission and causes behavioral defects. Neuron 14, 341–351 (1995).

  26. 26

    Kittel, R.J. et al. Bruchpilot promotes active zone assembly, Ca2+ channel clustering, and vesicle release. Science 312, 1051–1054 (2006).

  27. 27

    Wagh, D.A. et al. Bruchpilot, a protein with homology to ELKS/CAST, is required for structural integrity and function of synaptic active zones in Drosophila. Neuron 49, 833–844 (2006).

  28. 28

    Atwood, H.L. & Karunanithi, S. Diversification of synaptic strength: presynaptic elements. Nat. Rev. Neurosci. 3, 497–516 (2002).

  29. 29

    Barry, M.F. & Ziff, E.B. Receptor trafficking and the plasticity of excitatory synapses. Curr. Opin. Neurobiol. 12, 279–286 (2002).

  30. 30

    Malinow, R. & Malenka, R.C. AMPA receptor trafficking and synaptic plasticity. Annu. Rev. Neurosci. 25, 103–126 (2002).

  31. 31

    Bredt, D.S. & Nicoll, R.A. AMPA receptor trafficking at excitatory synapses. Neuron 40, 361–379 (2003).

  32. 32

    Zhong, Y., Budnik, V. & Wu, C.F. Synaptic plasticity in Drosophila memory and hyperexcitable mutants: role of cAMP cascade. J. Neurosci. 12, 644–651 (1992).

  33. 33

    Sigrist, S.J., Reiff, D.F., Thiel, P.R., Steinert, J.R. & Schuster, C.M. Experience-dependent strengthening of Drosophila neuromuscular junctions. J. Neurosci. 23, 6546–6556 (2003).

  34. 34

    Zhong, Y. & Wu, C.F. Neuronal activity and adenylyl cyclase in environment-dependent plasticity of axonal outgrowth in Drosophila. J. Neurosci. 24, 1439–1445 (2004).

  35. 35

    Bogdanik, L. et al. The Drosophila metabotropic glutamate receptor DmGluRA regulates activity-dependent synaptic facilitation and fine synaptic morphology. J. Neurosci. 24, 9105–9116 (2004).

  36. 36

    Featherstone, D.E. et al. Presynaptic glutamic acid decarboxylase is required for induction of the postsynaptic receptor field at a glutamatergic synapse. Neuron 27, 71–84 (2000).

  37. 37

    Ruiz-Canada, C. et al. New synaptic bouton formation is disrupted by misregulation of microtubule stability in aPKC mutants. Neuron 42, 567–580 (2004).

  38. 38

    Akaaboune, M., Grady, R.M., Turney, S., Sanes, J.R. & Lichtman, J.W. Neurotransmitter receptor dynamics studied in vivo by reversible photo-unbinding of fluorescent ligands. Neuron 34, 865–876 (2002).

  39. 39

    Schmid, A. et al. Non-NMDA-type glutamate receptors are essential for maturation, but not for initial assembly of synapses at Drosophila neuromuscular junctions. J. Neurosci. 26, 11267–11277 (2006).

  40. 40

    Chen, K., Merino, C., Sigrist, S.J. & Featherstone, D.E. The 4.1 protein coracle mediates subunit-selective anchoring of Drosophila glutamate receptors to the postsynaptic actin cytoskeleton. J. Neurosci. 25, 6667–6675 (2005).

  41. 41

    Ehlers, M.D., Heine, M., Groc, L., Lee, M.C. & Choquet, D. Diffusional trapping of GluR1 AMPA receptors by input-specific synaptic activity. Neuron 54, 447–460 (2007).

  42. 42

    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).

  43. 43

    Thiagarajan, T.C., Lindskog, M. & Tsien, R.W. Adaptation to synaptic inactivity in hippocampal neurons. Neuron 47, 725–737 (2005).

  44. 44

    Thiagarajan, T.C., Lindskog, M., Malgaroli, A. & Tsien, R.W. LTP and adaptation to inactivity: overlapping mechanisms and implications for metaplasticity. Neuropharmacology 52, 156–175 (2007).

  45. 45

    Holcman, D. & Triller, A. Modeling synaptic dynamics driven by receptor lateral diffusion. Biophys. J. 91, 2405–2415 (2006).

  46. 46

    Frank, C.A., Kennedy, M.J., Goold, C.P., Marek, K.W. & Davis, G.W. Mechanisms underlying the rapid induction and sustained expression of synaptic homeostasis. Neuron 52, 663–677 (2006).

  47. 47

    Verstreken, P. et al. Endophilin mutations block clathrin-mediated endocytosis, but not neurotransmitter release. Cell 109, 101–112 (2002).

  48. 48

    Bacci, A. et al. Chronic blockade of glutamate receptors enhances presynaptic release and downregulates the interaction between synaptophysin-synaptobrevin vesicle–associated membrane protein 2. J. Neurosci. 21, 6588–6596 (2001).

Download references

Acknowledgements

We would like to thank D.E. Featherstone for help with establishing patch-clamp recordings from Drosophila embryos and A. DiAntonio for fly stocks. This work was supported by grants from the Deutsche Forschungsgemeinschaft to S.J.S. (SI849/2-1 and 2-2, TP A16/SFB 406, TP B25/SFB581, SFB487) and to M.H. (HE 2621/4-1 and TP B22/SFB 581), and by formel.1 grants to S.H. and R.J.K. from the Medical Faculty of the University of Leipzig.

Author information

Correspondence to Stephan J Sigrist.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–3 and Methods (PDF 358 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Schmid, A., Hallermann, S., Kittel, R. et al. Activity-dependent site-specific changes of glutamate receptor composition in vivo. Nat Neurosci 11, 659–666 (2008) doi:10.1038/nn.2122

Download citation

Further reading