Abstract
Trafficking of AMPA receptors (AMPARs) is important for many forms of synaptic plasticity. However, the link between activity and resulting synaptic alterations is not fully understood. We identified a direct interaction between N-ethylmaleimide–sensitive fusion protein (NSF), an ATPase involved in membrane fusion events and stabilization of surface AMPARs, and Polo-like kinase- 2 (Plk2), an activity-inducible kinase that homeostatically decreases excitatory synapse number and strength. Plk2 disrupted the interaction of NSF with the GluA2 subunit of AMPARs, promoting extensive loss of surface GluA2 in rat hippocampal neurons, greater association of GluA2 with adaptor proteins PICK1 and GRIP1, and decreased synaptic AMPAR current. Plk2 engagement of NSF, but not Plk2 kinase activity, was required for this mechanism and occurred through a motif in the Plk2 protein that was independent of the canonical polo box interaction sites. These data reveal that heightened synaptic activity, acting through Plk2, leads to homeostatic decreases in surface AMPAR expression via the direct dissociation of NSF from GluA2.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Sheng, M. & Lee, S.H. AMPA receptor trafficking and the control of synaptic transmission. Cell 105, 825–828 (2001).
Zhao, C., Slevin, J.T. & Whiteheart, S.W. Cellular functions of NSF: Not just SNAPs and SNAREs. FEBS Lett. 581, 2140–2149 (2007).
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).
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).
Lüscher, C. et al. Role of AMPA receptor cycling in synaptic transmission and plasticity. Neuron 24, 649–658 (1999).
Braithwaite, S.P., Xia, H. & Malenka, R.C. Differential roles for NSF and GRIP/ABP in AMPA receptor cycling. Proc. Natl. Acad. Sci. USA 99, 7096–7101 (2002).
Shi, S., Hayashi, Y., Esteban, J.A. & Malinow, R. Subunit-specific rules governing AMPA receptor trafficking to synapses in hippocampal pyramidal neurons. Cell 105, 331–343 (2001).
Lüthi, A. et al. Hippocampal LTD expression involves a pool of AMPARs regulated by the NSF-GluR2 interaction. Neuron 24, 389–399 (1999).
Kim, C.H., Chung, H.J., Lee, H.K. & Huganir, R.L. Interaction of the AMPA receptor subunit GluR2/3 with PDZ domains regulates hippocampal long-term depression. Proc. Natl. Acad. Sci. USA 98, 11725–11730 (2001).
Noel, J. et al. Surface expression of AMPA receptors in hippocampal neurons is regulated by an NSF-dependent mechanism. Neuron 23, 365–376 (1999).
Simmons, D.L., Neel, B.G., Stevens, R., Evett, G. & Erikson, R.L. Identification of an early-growth-response gene encoding a novel putative protein kinase. Mol. Cell. Biol. 12, 4164–4169 (1992).
Kauselmann, G. et al. The polo-like protein kinases Fnk and Snk associate with a Ca2+- and integrin-binding protein and are regulated dynamically with synaptic plasticity. EMBO J. 18, 5528–5539 (1999).
Pak, D.T., Yang, S., Rudolph-Correia, S., Kim, E. & Sheng, M. Regulation of dendritic spine morphology by SPAR, a PSD-95–associated RapGAP. Neuron 31, 289–303 (2001).
Pak, D.T. & Sheng, M. Targeted protein degradation and synapse remodeling by an inducible protein kinase. Science 302, 1368–1373 (2003).
Ang, X.L., Seeburg, D.P., Sheng, M. & Harper, J.W. Regulation of postsynaptic RapGAP SPAR by Polo-like kinase 2 and the SCFbeta-TRCP ubiquitin ligase in hippocampal neurons. J. Biol. Chem. 283, 29424–29432 (2008).
Seeburg, D.P., Feliu-Mojer, M., Gaiottino, J., Pak, D.T. & Sheng, M. Critical role of CDK5 and Polo-like kinase 2 in homeostatic synaptic plasticity during elevated activity. Neuron 58, 571–583 (2008).
Seeburg, D.P. & Sheng, M. Activity-induced Polo-like kinase 2 is required for homeostatic plasticity of hippocampal neurons during epileptiform activity. J. Neurosci. 28, 6583–6591 (2008).
Elia, A.E. et al. The molecular basis for phosphodependent substrate targeting and regulation of Plks by the Polo-box domain. Cell 115, 83–95 (2003).
Feng, Y. et al. Association of polo-like kinase with alpha-, beta- and gamma-tubulins in a stable complex. Biochem. J. 339, 435–442 (1999).
Daw, M.I. et al. PDZ proteins interacting with C-terminal GluR2/3 are involved in a PKC-dependent regulation of AMPA receptors at hippocampal synapses. Neuron 28, 873–886 (2000).
Dong, H., Zhang, P., Liao, D. & Huganir, R.L. Characterization, expression, and distribution of GRIP protein. Ann. NY Acad. Sci. 868, 535–540 (1999).
Perez, J.L. et al. PICK1 targets activated protein kinase Cα to AMPA receptor clusters in spines of hippocampal neurons and reduces surface levels of the AMPA-type glutamate receptor subunit 2. J. Neurosci. 21, 5417–5428 (2001).
States, B.A., Khatri, L. & Ziff, E.B. Stable synaptic retention of serine-880–phosphorylated GluR2 in hippocampal neurons. Mol. Cell. Neurosci. 38, 189–202 (2008).
Lee, S.H., Liu, L., Wang, Y.T. & Sheng, M. Clathrin adaptor AP2 and NSF interact with overlapping sites of GluR2 and play distinct roles in AMPA receptor trafficking and hippocampal LTD. Neuron 36, 661–674 (2002).
Lee, K.S., Grenfell, T.Z., Yarm, F.R. & Erikson, R.L. Mutation of the polo-box disrupts localization and mitotic functions of the mammalian polo kinase Plk. Proc. Natl. Acad. Sci. USA 95, 9301–9306 (1998).
Turrigiano, G. Homeostatic signaling: the positive side of negative feedback. Curr. Opin. Neurobiol. 17, 318–324 (2007).
Gainey, M.A., Hurvitz-Wolff, J.R., Lambo, M.E. & Turrigiano, G.G. Synaptic scaling requires the GluR2 subunit of the AMPA receptor. J. Neurosci. 29, 6479–6489 (2009).
Lu, W. et al. Subunit composition of synaptic AMPA receptors revealed by a single-cell genetic approach. Neuron 62, 254–268 (2009).
Wenthold, R.J., Petralia, R.S., Blahos, J., II & Niedzielski, A.S. Evidence for multiple AMPA receptor complexes in hippocampal CA1/CA2 neurons. J. Neurosci. 16, 1982–1989 (1996).
Shepherd, J.D. et al. Arc/Arg3.1 mediates homeostatic synaptic scaling of AMPA receptors. Neuron 52, 475–484 (2006).
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).
Plant, K. et al. Transient incorporation of native GluR2-lacking AMPA receptors during hippocampal long-term potentiation. Nat. Neurosci. 9, 602–604 (2006).
Liu, B. et al. Ischemic insults direct glutamate receptor subunit 2-lacking AMPA receptors to synaptic sites. J. Neurosci. 26, 5309–5319 (2006).
Jia, Z. et al. Enhanced LTP in mice deficient in the AMPA receptor GluR2. Neuron 17, 945–956 (1996).
Meng, Y., Zhang, Y. & Jia, Z. Synaptic transmission and plasticity in the absence of AMPA glutamate receptor GluR2 and GluR3. Neuron 39, 163–176 (2003).
Biou, V., Bhattacharyya, S. & Malenka, R.C. Endocytosis and recycling of AMPA receptors lacking GluR2/3. Proc. Natl. Acad. Sci. USA 105, 1038–1043 (2008).
Gardner, S.M. et al. Calcium-permeable AMPA receptor plasticity is mediated by subunit-specific interactions with PICK1 and NSF. Neuron 45, 903–915 (2005).
Beretta, F. et al. NSF interaction is important for direct insertion of GluR2 at synaptic sites. Mol. Cell. Neurosci. 28, 650–660 (2005).
Narisawa-Saito, M. et al. Brain-derived neurotrophic factor regulates surface expression of α-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid receptors by enhancing the N-ethylmaleimide–sensitive factor/GluR2 interaction in developing neocortical neurons. J. Biol. Chem. 277, 40901–40910 (2002).
Lin, D.T. & Huganir, R.L. PICK1 and phosphorylation of the glutamate receptor 2 (GluR2) AMPA receptor subunit regulates GluR2 recycling after NMDA receptor–induced internalization. J. Neurosci. 27, 13903–13908 (2007).
Yao, Y. et al. PKM zeta maintains late long-term potentiation by N-ethylmaleimide–sensitive factor/GluR2-dependent trafficking of postsynaptic AMPA receptors. J. Neurosci. 28, 7820–7827 (2008).
Huang, Y. et al. S-nitrosylation of N-ethylmaleimide–sensitive factor mediates surface expression of AMPA receptors. Neuron 46, 533–540 (2005).
Hanley, J.G., Khatri, L., Hanson, P.I. & Ziff, E.B. NSF ATPase and alpha-/beta-SNAPs disassemble the AMPA receptor–PICK1 complex. Neuron 34, 53–67 (2002).
Xia, J., Chung, H.J., Wihler, C., Huganir, R.L. & Linden, D.J. Cerebellar long-term depression requires PKC-regulated interactions between GluR2/3 and PDZ domain–containing proteins. Neuron 28, 499–510 (2000).
Terashima, A. et al. Regulation of synaptic strength and AMPA receptor subunit composition by PICK1. J. Neurosci. 24, 5381–5390 (2004).
Osten, P. et al. Mutagenesis reveals a role for ABP/GRIP binding to GluR2 in synaptic surface accumulation of the AMPA receptor. Neuron 27, 313–325 (2000).
DeSouza, S., Fu, J., States, B.A. & Ziff, E.B. Differential palmitoylation directs the AMPA receptor–binding protein ABP to spines or to intracellular clusters. J. Neurosci. 22, 3493–3503 (2002).
Chung, H.J., Xia, J., Scannevin, R.H., Zhang, X. & Huganir, R.L. Phosphorylation of the AMPA receptor subunit GluR2 differentially regulates its interaction with PDZ domain–containing proteins. J. Neurosci. 20, 7258–7267 (2000).
Fukata, Y. et al. Molecular constituents of neuronal AMPA receptors. J. Cell Biol. 169, 399–404 (2005).
Acknowledgements
We thank L. Bilello for technical assistance. This work was supported by US National Institutes of Health and National Institute of Neurological Disorders and Stroke grants NS048085 (D.T.S.P.), F31NS061467 (D.M.E.) and P30CA051008 (Biacore Molecular Interactions Shared Resource of Lombardi Comprehensive Cancer Center).
Author information
Authors and Affiliations
Contributions
D.M.E., H.-S.H., J.A.M., D.Z. and D.T.S.P. performed experiments. D.M.E., S.H.L., J.T.I. and D.T.S.P. designed the experiments. D.M.E. and D.T.S.P. wrote the manuscript. D.T.S.P. supervised the project.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–9 and SupplementaryTables 1 and 2 (PDF 1159 kb)
Rights and permissions
About this article
Cite this article
Evers, D., Matta, J., Hoe, HS. et al. Plk2 attachment to NSF induces homeostatic removal of GluA2 during chronic overexcitation. Nat Neurosci 13, 1199–1207 (2010). https://doi.org/10.1038/nn.2624
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nn.2624
This article is cited by
-
A Highly Polymorphic Copy Number Variant in the NSF Gene is Associated with Cocaine Dependence
Scientific Reports (2016)
-
N-ethylmaleimide-sensitive factor interacts with the serotonin transporter and modulates its trafficking: implications for pathophysiology in autism
Molecular Autism (2014)
-
AMPAR trafficking in synapse maturation and plasticity
Cellular and Molecular Life Sciences (2013)
-
GKAP orchestrates activity-dependent postsynaptic protein remodeling and homeostatic scaling
Nature Neuroscience (2012)