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A critical role for PSD-95/AKAP interactions in endocytosis of synaptic AMPA receptors

Abstract

The endocytosis of AMPA receptors (AMPARs) underlies several forms of synaptic plasticity, including NMDA receptor (NMDAR)-dependent long-term depression (LTD), but the molecular mechanisms responsible for this trafficking remain unknown. We found that PSD-95, a major postsynaptic density protein, is important for NMDAR-triggered endocytosis of synaptic AMPARs in rat neuron cultures because of its binding to A kinase–anchoring protein 150 (AKAP150), a scaffold for specific protein kinases and phosphatases. Knockdown of PSD-95 with shRNA blocked NMDAR-triggered, but not constitutive or mGluR-triggered, endocytosis of AMPARs. Deletion of PSD-95's Src homology 3 and guanylate kinase–like domains, as well as a point mutation (L460P), both of which inhibit binding of PSD-95 to AKAP150, also blocked NMDAR-triggered AMPAR endocytosis. Furthermore, expression of a mutant AKAP150 that does not bind calcineurin inhibited this NMDAR-triggered trafficking event. Our results suggest that PSD-95's interaction with AKAP150 is critical for NMDAR-triggered AMPAR endocytosis and LTD, possibly because these scaffolds position calcineurin in the appropriate subsynaptic domain.

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Figure 1: NMDAR- and AMPAR-triggered endocytosis of AMPARs show distinct patterns of internalization.
Figure 2: Acute knockdown of PSD-95 decreases the surface expression and NMDAR-triggered endocytosis of AMPARs.
Figure 3: Constitutive endocytosis and mGluR-triggered endocytosis of AMPARs are not affected by knockdown of PSD-95.
Figure 4: PEST motif deletion and prenylation of PSD-95 does not affect NMDAR-triggered endocytosis of AMPARs.
Figure 5: SH3 and GK domains of PSD-95 are required for NMDAR-triggered endocytosis of AMPARs.
Figure 6: L460P mutation in PSD-95 disrupts binding to AKAP150 and blocks NMDAR-triggered endocytosis of AMPARs.
Figure 7: Binding of calcineurin (PP2B) to AKAP150 is necessary for the NMDAR-triggered endocytosis of AMPARs.
Figure 8: NMDA application causes loss of PSD-95 and AKAP150 from synapses.

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References

  1. Malenka, R.C. & Bear, M.F. LTP and LTD: an embarrassment of riches. Neuron 44, 5–21 (2004).

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  3. Collingridge, G.L., Isaac, J.T. & Wang, Y.T. Receptor trafficking and synaptic plasticity. Nat. Rev. Neurosci. 5, 952–962 (2004).

    Article  CAS  PubMed  Google Scholar 

  4. Shepherd, J.D. & Huganir, R.L. The cell biology of synaptic plasticity: AMPA receptor trafficking. Annu. Rev. Cell Dev. Biol. 23, 613–643 (2007).

    Article  CAS  PubMed  Google Scholar 

  5. Hollmann, M. & Heinemann, S. Cloned glutamate receptors. Annu. Rev. Neurosci. 17, 31–108 (1994).

    Article  CAS  PubMed  Google Scholar 

  6. Rosenmund, C., Stern-Bach, Y. & Stevens, C.F. The tetrameric structure of a glutamate receptor channel. Science 280, 1596–1599 (1998).

    Article  CAS  PubMed  Google Scholar 

  7. Kim, E. & Sheng, M. PDZ domain proteins of synapses. Nat. Rev. Neurosci. 5, 771–781 (2004).

    Article  CAS  PubMed  Google Scholar 

  8. Nicoll, R.A., Tomita, S. & Bredt, D.S. Auxiliary subunits assist AMPA-type glutamate receptors. Science 311, 1253–1256 (2006).

    Article  CAS  PubMed  Google Scholar 

  9. El-Husseini, A.E., Schnell, E., Chetkovich, D.M., Nicoll, R.A. & Bredt, D.S. PSD-95 involvement in maturation of excitatory synapses. Science 290, 1364–1368 (2000).

    CAS  PubMed  Google Scholar 

  10. Ehrlich, I. & Malinow, R. Postsynaptic density 95 controls AMPA receptor incorporation during long-term potentiation and experience-driven synaptic plasticity. J. Neurosci. 24, 916–927 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Schluter, O.M., Xu, W. & Malenka, R.C. Alternative N-terminal domains of PSD-95 and SAP97 govern activity-dependent regulation of synaptic AMPA receptor function. Neuron 51, 99–111 (2006).

    Article  CAS  PubMed  Google Scholar 

  12. Schnell, E. et al. Direct interactions between PSD-95 and stargazin control synaptic AMPA receptor number. Proc. Natl. Acad. Sci. USA 99, 13902–13907 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Nakagawa, T. et al. Quaternary structure, protein dynamics and synaptic function of SAP97 controlled by L27 domain interactions. Neuron 44, 453–467 (2004).

    Article  CAS  PubMed  Google Scholar 

  14. Ehrlich, I., Klein, M., Rumpel, S. & Malinow, R. PSD-95 is required for activity-driven synapse stabilization. Proc. Natl. Acad. Sci. USA 104, 4176–4181 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Elias, G.M. et al. Synapse-specific and developmentally regulated targeting of AMPA receptors by a family of MAGUK scaffolding proteins. Neuron 52, 307–320 (2006).

    CAS  PubMed  Google Scholar 

  16. Futai, K. et al. Retrograde modulation of presynaptic release probability through signaling mediated by PSD-95-neuroligin. Nat. Neurosci. 10, 186–195 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Stein, V., House, D.R., Bredt, D.S. & Nicoll, R.A. Postsynaptic density–95 mimics and occludes hippocampal long-term potentiation and enhances long-term depression. J. Neurosci. 23, 5503–5506 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Colledge, M. et al. Ubiquitination regulates PSD-95 degradation and AMPA receptor surface expression. Neuron 40, 595–607 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. El-Husseini, A.D. et al. Synaptic strength regulated by palmitate cycling on PSD-95. Cell 108, 849–863 (2002).

    Article  CAS  Google Scholar 

  20. Beattie, E.C. et al. Regulation of AMPA receptor endocytosis by a signaling mechanism shared with LTD. Nat. Neurosci. 3, 1291–1300 (2000).

    Article  CAS  PubMed  Google Scholar 

  21. Carroll, R.C. et al. Dynamin-dependent endocytosis of ionotropic glutamate receptors. Proc. Natl. Acad. Sci. USA 96, 14112–14117 (1999).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Lin, J.W. et al. Distinct molecular mechanisms and divergent endocytotic pathways of AMPA receptor internalization. Nat. Neurosci. 3, 1282–1290 (2000).

    Article  CAS  PubMed  Google Scholar 

  23. Xu, W. et al. Molecular dissociation of the role of PSD-95 in regulating synaptic strength and LTD. Neuron 57, 248–262 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Beene, D.L. & Scott, J.D. A-kinase anchoring proteins take shape. Curr. Opin. Cell Biol. 19, 192–198 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Dell'Acqua, M.L. et al. Regulation of neuronal PKA signaling through AKAP targeting dynamics. Eur. J. Cell Biol. 85, 627–633 (2006).

    Article  CAS  PubMed  Google Scholar 

  26. Klauck, T.M. et al. Coordination of three signaling enzymes by AKAP79, a mammalian scaffold protein. Science 271, 1589–1592 (1996).

    Article  CAS  PubMed  Google Scholar 

  27. Mulkey, R.M., Endo, S., Shenolikar, S. & Malenka, R.C. Involvement of a calcineurin/inhibitor-1 phosphatase cascade in hippocampal long-term depression. Nature 369, 486–488 (1994).

    Article  CAS  PubMed  Google Scholar 

  28. Snyder, E.M. et al. Role for A kinase-anchoring proteins (AKAPS) in glutamate receptor trafficking and long term synaptic depression. J. Biol. Chem. 280, 16962–16968 (2005).

    Article  CAS  PubMed  Google Scholar 

  29. Ehlers, M.D. Reinsertion or degradation of AMPA receptors determined by activity-dependent endocytic sorting. Neuron 28, 511–525 (2000).

    Article  CAS  PubMed  Google Scholar 

  30. Snyder, E.M. et al. Internalization of ionotropic glutamate receptors in response to mGluR activation. Nat. Neurosci. 4, 1079–1085 (2001).

    Article  CAS  PubMed  Google Scholar 

  31. Lee, S.H., Simonetta, A. & Sheng, M. Subunit rules governing the sorting of internalized AMPA receptors in hippocampal neurons. Neuron 43, 221–236 (2004).

    Article  CAS  PubMed  Google Scholar 

  32. Regalado, M.P., Terry-Lorenzo, R.T., Waites, C.L., Garner, C.C. & Malenka, R.C. Transsynaptic signaling by postsynaptic synapse-associated protein 97. J. Neurosci. 26, 2343–2357 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Colledge, M. et al. Targeting of PKA to glutamate receptors through a MAGUK-AKAP complex. Neuron 27, 107–119 (2000).

    Article  CAS  PubMed  Google Scholar 

  34. McGee, A.W. & Bredt, D.S. Identification of an intramolecular interaction between the SH3 and guanylate kinase domains of PSD-95. J. Biol. Chem. 274, 17431–17436 (1999).

    Article  CAS  PubMed  Google Scholar 

  35. Hoshi, N., Langeberg, L.K. & Scott, J.D. Distinct enzyme combinations in AKAP signaling complexes permit functional diversity. Nat. Cell Biol. 7, 1066–1073 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Dell'Acqua, M.L., Dodge, K.L., Tavalin, S.J. & Scott, J.D. Mapping the protein phosphatase 2B anchoring site on AKAP79. Binding and inhibition of phosphatase activity are mediated by residues 315–360. J. Biol. Chem. 277, 48796–48802 (2002).

    Article  CAS  PubMed  Google Scholar 

  37. Smith, K.E., Gibson, E.S. & Dell'Acqua, M.L. cAMP-dependent protein kinase postsynaptic localization regulated by NMDA receptor activation through translocation of an A-kinase anchoring protein scaffold protein. J. Neurosci. 26, 2391–2402 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Cummings, J.A., Mulkey, R.M., Nicoll, R.A. & Malenka, R.C. Ca2+ signaling requirements for long-term depression in the hippocampus. Neuron 16, 825–833 (1996).

    Article  CAS  PubMed  Google Scholar 

  39. Oliveria, S.F., Dell'Acqua, M.L. & Sather, W.A. AKAP79/150 anchoring of calcineurin controls neuronal L-type Ca2+ channel activity and nuclear signaling. Neuron 55, 261–275 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Coghlan, V.M. et al. Association of protein kinase A and protein phosphatase 2B with a common anchoring protein. Science 267, 108–111 (1995).

    Article  CAS  PubMed  Google Scholar 

  41. Kashishian, A. et al. AKAP79 inhibits calcineurin through a site distinct from the immunophilin-binding region. J. Biol. Chem. 273, 27412–27419 (1998).

    Article  CAS  PubMed  Google Scholar 

  42. Gomez, L.L., Alam, S., Smith, K.E., Horne, E. & Dell'Acqua, M.L. Regulation of A-kinase anchoring protein 79/150-cAMP–dependent protein kinase postsynaptic targeting by NMDA receptor activation of calcineurin and remodeling of dendritic actin. J. Neurosci. 22, 7027–7044 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Tavalin, S.J. et al. Regulation of GluR1 by the A-kinase anchoring protein 79 (AKAP79) signaling complex shares properties with long-term depression. J. Neurosci. 22, 3044–3051 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Tomita, S., Stein, V., Stocker, T.J., Nicoll, R.A. & Bredt, D.S. Bidirectional synaptic plasticity regulated by phosphorylatoin of stargazin-like TARPs. Neuron 45, 269–277 (2005).

    Article  CAS  PubMed  Google Scholar 

  45. Tomita, S., Fukata, M., Nicoll, R.A. & Bredt, D.S. Dynamic interaction of stargazin-like TARPs with cycling AMPA receptors at synapses. Science 303, 1508–1511 (2004).

    Article  CAS  PubMed  Google Scholar 

  46. Gallagher, S.M., Daly, C.A., Bear, M.F. & Huber, K.M. Extracellular signal–regulated protein kinase activation is required for metabotropic glutamate receptor–dependent long-term depression in hippocampal area CA1. J. Neurosci. 24, 4859–4864 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Schnabel, R., Kilpatrick, I.C. & Collingridge, G.L. Protein phosphatase inhibitors facilitate DHPG-induced LTD in the CA1 region of the hippocampus. Br. J. Pharmacol. 132, 1095–1101 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Davidkova, G. & Carroll, R.C. Characterization of the role of microtubule-associated protein 1B in metabotropic glutamate receptor–mediated endocytosis of AMPA receptors in hippocampus. J. Neurosci. 27, 13273–13278 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Ronesi, J.A. & Huber, K.M. Homer interactions are necessary for metabotropic glutamate receptor–induced long-term depression and translational activation. J. Neurosci. 28, 543–547 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank A. Ghosh and X. Cai for expert technical assistance and members of the Malenka laboratory for helpful discussions. This work was supported by US National Institutes of Health grant MH63394 (to R.C.M.) and MH080310 (to W.X.) and postdoctoral fellowship SCHL592/4-1 from the DFG (to O.S.).

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All authors contributed to the design and interpretation of the experiments and commented on the manuscript. S.B. conducted and analyzed all experiments. S.B., V.B., W.X. and O.S. prepared constructs. S.B. and R.C.M. wrote the manuscript.

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Correspondence to Robert C Malenka.

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Bhattacharyya, S., Biou, V., Xu, W. et al. A critical role for PSD-95/AKAP interactions in endocytosis of synaptic AMPA receptors. Nat Neurosci 12, 172–181 (2009). https://doi.org/10.1038/nn.2249

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