Abstract
Regulation of calcium-permeable AMPA receptors (CP-AMPARs) is crucial in normal synaptic function and neurological disease states. Although transmembrane AMPAR regulatory proteins (TARPs) such as stargazin (γ-2) modulate the properties of calcium-impermeable AMPARs (CI-AMPARs) and promote their synaptic targeting, the TARP-specific rules governing CP-AMPAR synaptic trafficking remain unclear. We used RNA interference to manipulate AMPAR-subunit and TARP expression in γ-2–lacking stargazer cerebellar granule cells—the classic model of TARP deficiency. We found that TARP γ-7 selectively enhanced the synaptic expression of CP-AMPARs and suppressed CI-AMPARs, identifying a pivotal role of γ-7 in regulating the prevalence of CP-AMPARs. In the absence of associated TARPs, both CP-AMPARs and CI-AMPARs were able to localize to synapses and mediate transmission, although their properties were altered. Our results also establish that TARPed synaptic receptors in granule cells require both γ-2 and γ-7 and reveal an unexpected basis for the loss of AMPAR-mediated transmission in stargazer mice.
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
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Accession codes
References
Traynelis, S.F. et al. Glutamate receptor ion channels: structure, regulation, and function. Pharmacol. Rev. 62, 405–496 (2010).
Kuner, R. et al. Late-onset motoneuron disease caused by a functionally modified AMPA receptor subunit. Proc. Natl. Acad. Sci. USA 102, 5826–5831 (2005).
Liu, B. et al. Ischemic insults direct glutamate receptor subunit 2–lacking AMPA receptors to synaptic sites. J. Neurosci. 26, 5309–5319 (2006).
Park, J.S. et al. Persistent inflammation induces GluR2 internalization via NMDA receptor–triggered PKC activation in dorsal horn neurons. J. Neurosci. 29, 3206–3219 (2009).
Gangadharan, V. et al. Peripheral calcium-permeable AMPA receptors regulate chronic inflammatory pain in mice. J. Clin. Invest. 121, 1608–1623 (2011).
Sommer, B., Kohler, M., Sprengel, R. & Seeburg, P.H. RNA editing in brain controls a determinant of ion flow in glutamate-gated channels. Cell 67, 11–19 (1991).
Rosenthal, J.J. & Seeburg, P.H. A-to-I RNA editing: effects on proteins key to neural excitability. Neuron 74, 432–439 (2012).
Hollmann, M., Hartley, M. & Heinemann, S. Ca2+ permeability of KA-AMPA–gated glutamate receptor channels depends on subunit composition. Science 252, 851–853 (1991).
Burnashev, N., Monyer, H., Seeburg, P.H. & Sakmann, B. Divalent ion permeability of AMPA receptor channels is dominated by the edited form of a single subunit. Neuron 8, 189–198 (1992).
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).
Cull-Candy, S., Kelly, L. & Farrant, M. Regulation of Ca2+-permeable AMPA receptors: synaptic plasticity and beyond. Curr. Opin. Neurobiol. 16, 288–297 (2006).
Isaac, J.T., Ashby, M. & McBain, C.J. The role of the GluR2 subunit in AMPA receptor function and synaptic plasticity. Neuron 54, 859–871 (2007).
Liu, S.J. & Zukin, R.S. Ca2+-permeable AMPA receptors in synaptic plasticity and neuronal death. Trends Neurosci. 30, 126–134 (2007).
Bowie, D., Lange, G.D. & Mayer, M.L. Activity-dependent modulation of glutamate receptors by polyamines. J. Neurosci. 18, 8175–8185 (1998).
Rozov, A. & Burnashev, N. Polyamine-dependent facilitation of postsynaptic AMPA receptors counteracts paired-pulse depression. Nature 401, 594–598 (1999).
Liu, S.Q.J. & Cull-Candy, S.G. Synaptic activity at calcium-permeable AMPA receptors induces a switch in receptor subtype. Nature 405, 454–458 (2000).
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).
Man, H.Y. GluA2-lacking, calcium-permeable AMPA receptors—inducers of plasticity? Curr. Opin. Neurobiol. 21, 291–298 (2011).
Priel, A. et al. Stargazin reduces desensitization and slows deactivation of the AMPA-type glutamate receptors. J. Neurosci. 25, 2682–2686 (2005).
Tomita, S. et al. Stargazin modulates AMPA receptor gating and trafficking by distinct domains. Nature 435, 1052–1058 (2005).
Turetsky, D., Garringer, E. & Patneau, D.K. Stargazin modulates native AMPA receptor functional properties by two distinct mechanisms. J. Neurosci. 25, 7438–7448 (2005).
Bedoukian, M.A., Weeks, A.M. & Partin, K.M. Different domains of the AMPA receptor direct stargazin-mediated trafficking and stargazin-mediated modulation of kinetics. J. Biol. Chem. 281, 23908–23921 (2006).
Cho, C.H., St-Gelais, F., Zhang, W., Tomita, S. & Howe, J.R. Two families of TARP isoforms that have distinct effects on the kinetic properties of AMPA receptors and synaptic currents. Neuron 55, 890–904 (2007).
Kott, S., Sager, C., Tapken, D., Werner, M. & Hollmann, M. Comparative analysis of the pharmacology of GluR1 in complex with transmembrane AMPA receptor regulatory proteins γ 2, γ3, γ4, and γ 8. Neuroscience 158, 78–88 (2009).
Tomita, S. et al. Functional studies and distribution define a family of transmembrane AMPA receptor regulatory proteins. J. Cell Biol. 161, 805–816 (2003).
Kato, A.S. et al. New transmembrane AMPA receptor regulatory protein isoform, γ-7, differentially regulates AMPA receptors. J. Neurosci. 27, 4969–4977 (2007).
Soto, D. et al. Selective regulation of long-form calcium-permeable AMPA receptors by an atypical TARP, γ-5. Nat. Neurosci. 12, 277–285 (2009).
Fukaya, M., Yamazaki, M., Sakimura, K. & Watanabe, M. Spatial diversity in gene expression for VDCCγ subunit family in developing and adult mouse brains. Neurosci. Res. 53, 376–383 (2005).
Yamazaki, M. et al. TARPs γ-2 and γ-7 are essential for AMPA receptor expression in the cerebellum. Eur. J. Neurosci. 31, 2204–2220 (2010).
Jackson, A.C. & Nicoll, R.A. Stargazin (TARP γ-2) is required for compartment-specific AMPA receptor trafficking and synaptic plasticity in cerebellar stellate cells. J. Neurosci. 31, 3939–3952 (2011).
Bats, C., Soto, D., Studniarczyk, D., Farrant, M. & Cull-Candy, S.G. Channel properties reveal differential expression of TARPed and TARPless AMPARs in stargazer neurons. Nat. Neurosci. 15, 853–861 (2012).
Hashimoto, K. et al. Impairment of AMPA receptor function in cerebellar granule cells of ataxic mutant mouse stargazer. J. Neurosci. 19, 6027–6036 (1999).
Chen, L. et al. Stargazin regulates synaptic targeting of AMPA receptors by two distinct mechanisms. Nature 408, 936–943 (2000).
Bats, C., Groc, L. & Choquet, D. The interaction between Stargazin and PSD-95 regulates AMPA receptor surface trafficking. Neuron 53, 719–734 (2007).
Gallo, V. et al. Molecular cloning and development analysis of a new glutamate receptor subunit isoform in cerebellum. J. Neurosci. 12, 1010–1023 (1992).
Mosbacher, J. et al. A molecular determinant for submillisecond desensitization in glutamate receptors. Science 266, 1059–1062 (1994).
Cathala, L., Holderith, N.B., Nusser, Z., DiGregorio, D.A. & Cull-Candy, S.G. Changes in synaptic structure underlie the developmental speeding of AMPA receptor–mediated EPSCs. Nat. Neurosci. 8, 1310–1318 (2005).
Schwenk, J. et al. Functional proteomics identify cornichon proteins as auxiliary subunits of AMPA receptors. Science 323, 1313–1319 (2009).
Shi, Y. et al. Functional comparison of the effects of TARPs and cornichons on AMPA receptor trafficking and gating. Proc. Natl. Acad. Sci. USA 107, 16315–16319 (2010).
Soto, D., Coombs, I.D., Kelly, L., Farrant, M. & Cull-Candy, S.G. Stargazin attenuates intracellular polyamine block of calcium-permeable AMPA receptors. Nat. Neurosci. 10, 1260–1267 (2007).
Washburn, M.S. & Dingledine, R. Block of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors by polyamines and polyamine toxins. J. Pharmacol. Exp. Ther. 278, 669–678 (1996).
Koike, M., Iino, M. & Ozawa, S. Blocking effect of 1-naphthyl acetyl spermine on Ca2+-permeable AMPA receptors in cultured rat hippocampal neurons. Neurosci. Res. 29, 27–36 (1997).
Blaschke, M. et al. A single amino acid determines the subunit-specific spider toxin block of α-amino-3-hydroxy-5-methylisoxazole-4-propionate/kainate receptor channels. Proc. Natl. Acad. Sci. USA 90, 6528–6532 (1993).
Menuz, K., O'Brien, J.L., Karmizadegan, S., Bredt, D.S. & Nicoll, R.A. TARP redundancy is critical for maintaining AMPA receptor function. J. Neurosci. 28, 8740–8746 (2008).
Kato, A.S., Siuda, E.R., Nisenbaum, E.S. & Bredt, D.S. AMPA receptor subunit–specific regulation by a distinct family of type II TARPs. Neuron 59, 986–996 (2008).
Rouach, N. et al. TARP γ-8 controls hippocampal AMPA receptor number, distribution and synaptic plasticity. Nat. Neurosci. 8, 1525–1533 (2005).
Shi, Y., Lu, W., Milstein, A.D. & Nicoll, R.A. The stoichiometry of AMPA receptors and TARPs varies by neuronal cell type. Neuron 62, 633–640 (2009).
Kim, K.S., Yan, D. & Tomita, S. Assembly and stoichiometry of the AMPA receptor and transmembrane AMPA receptor regulatory protein complex. J. Neurosci. 30, 1064–1072 (2010).
Yan, D. & Tomita, S. Defined criteria for auxiliary subunits of glutamate receptors. J. Physiol. (Lond.) 590, 21–31 (2012).
Letts, V.A. et al. The mouse stargazer gene encodes a neuronal Ca2+-channel γ subunit. Nat. Genet. 19, 340–347 (1998).
Passafaro, M., Nakagawa, T., Sala, C. & Sheng, M. Induction of dendritic spines by an extracellular domain of AMPA receptor subunit GluR2. Nature 424, 677–681 (2003).
Schneider, C.A., Rasband, W.S. & Eliceiri, K.W. NIH Image to ImageJ: 25 years of image analysis. Nat. Methods 9, 671–675 (2012).
Ferron, L. et al. The stargazin-related protein γ 7 interacts with the mRNA-binding protein heterogeneous nuclear ribonucleoprotein A2 and regulates the stability of specific mRNAs, including CaV2.2. J. Neurosci. 28, 10604–10617 (2008).
Chung, C., Deak, F. & Kavalali, E.T. Molecular substrates mediating lanthanide-evoked neurotransmitter release in central synapses. J. Neurophysiol. 100, 2089–2100 (2008).
Kudoh, S.N. & Taguchi, T. A simple exploratory algorithm for the accurate and fast detection of spontaneous synaptic events. Biosens. Bioelectron. 17, 773–782 (2002).
Traynelis, S.F., Silver, R.A. & Cull-Candy, S.G. Estimated conductance of glutamate receptor channels activated during EPSCs at the cerebellar mossy fiber-granule cell synapse. Neuron 11, 279–289 (1993).
Wilcox, R.R. Introduction to Robust Estimation and Hypothesis Testing (Academic Press, Amsterdam, Boston, 2012).
Maechler, M., Rousseeuw, P., Struyf, A., Hubert, M. & Hornik, K. Cluster analysis basics and extensions. R package version 1.14.3. (2012).
Acknowledgements
This work was supported by Programme Grants from the Wellcome Trust and the Medical Research Council (S.G.C.-C. and M.F.). We thank M. Watanabe for antibodies against γ-2 and γ-7, R. Nicoll for TARP γ-2 cDNA, C. Bats for invaluable discussions and M. Zonouzi and M. Watanabe for advice on coimmunoprecipitation.
Author information
Authors and Affiliations
Contributions
S.G.C.-C. and M.F. conceived and supervised the project. D.S. performed electrophysiological and biochemical experiments. M.F. and D.S. analyzed the data. I.C. generated reagents for RNA interference and performed biochemical experiments. D.S., S.G.C.-C. and M.F. wrote the manuscript.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Figures and Text
Supplementary Figures 1–3 and Supplementary Tables 1–4 (PDF 1043 kb)
Rights and permissions
About this article
Cite this article
Studniarczyk, D., Coombs, I., Cull-Candy, S. et al. TARP γ-7 selectively enhances synaptic expression of calcium-permeable AMPARs. Nat Neurosci 16, 1266–1274 (2013). https://doi.org/10.1038/nn.3473
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nn.3473
This article is cited by
-
Exploring the molecular basis of neuronal excitability in a vocal learner
BMC Genomics (2019)
-
AMPA receptors and their minions: auxiliary proteins in AMPA receptor trafficking
Cellular and Molecular Life Sciences (2019)
-
Retinal progenitor cells release extracellular vesicles containing developmental transcription factors, microRNA and membrane proteins
Scientific Reports (2018)
-
Synaptic AMPA receptor composition in development, plasticity and disease
Nature Reviews Neuroscience (2016)
-
Superactivation of AMPA receptors by auxiliary proteins
Nature Communications (2016)