Research Article
No access
Published Online: 18 March 2016

Pharmacological Modulation of GluK1 and GluK2 by NETO1, NETO2, and PSD95

Publication: ASSAY and Drug Development Technologies
Volume 14, Issue Number 2

Abstract

The association between the kainate receptors (KARs) GluK1 and GluK2 and the modifying proteins neuropilin- and tolloid-like 1 (NETO1), neuropilin- and tolloid-like 2 (NETO2), and postsynaptic density protein 95 (PSD95) is likely to produce distinct GluK1 and GluK2 pharmacology in postsynaptic neurons. However, little is known about their corresponding modulatory effects on GluK1 and GluK2 activity in high-throughput assays for cell-based drug discovery. Using heterologous cells that potentially mimic the response in native cells in a fluorescence imaging plate reader (FLIPR) assay, we have investigated assays that incorporate (1) coexpression of GluK1 or GluK2 with their modulatory proteins (NETO1, NETO2, PSD95) and/or (2) enablement of assays with physiological concentration of native GluK1 and GluK2 agonist (glutamate) in the absence of an artificial potentiator (e.g., concanavalin A [Con A]). We found that in the absence of Con A, both NETO1 and NETO2 accessory proteins are able to potentiate kainate- and glutamate-evoked GluK1-mediated Ca2+ influx. We also noted the striking ability of PSD95 to enhance glutamate-stimulated potentiation effects of NETO2 on GluK1 without the need for Con A and with a robust signal that could be utilized for high-throughput FLIPR assays. These experiments demonstrate the utility of heterologous cells coexpressing PSD95/NETO2 with GluK1 or GluK2 in native cell-mimicking heterologous cell systems for high-throughput assays and represent new avenues into the discovery of KAR modulating therapies.

Get full access to this article

View all available purchase options and get full access to this article.

References

1.
Dingledine R, Borges K, Bowie D, Traynelis SF: The glutamate receptor ion channels. Pharmacol Rev 1999;51:7–61.
2.
Kato AS, Gill MB, Ho MT, et al.: Hippocampal AMPA receptor gating controlled by both TARP and cornichon proteins. Neuron 2010;68:1082–1096.
3.
Schwenk J, Harmel N, Zolles G, et al.: Functional proteomics identify cornichon proteins as auxiliary subunits of AMPA receptors. Science 2009;323:1313–1319.
4.
Tomita S, Chen L, Kawasaki Y, et al.: Functional studies and distribution define a family of transmembrane AMPA receptor regulatory proteins. J Cell Biol 2003;161:805–816.
5.
Tomita S, Castillo PE: Neto1 and Neto2: auxiliary subunits that determine key properties of native kainate receptors. J Physiol 2012;590:2217–2223.
6.
Garcia EP, Mehta S, Blair LA, et al.: SAP90 binds and clusters kainate receptors causing incomplete desensitization. Neuron 1998;21:727–739.
7.
Bowie D, Garcia EP, Marshall J, Traynelis SF, Lange GD: Allosteric regulation and spatial distribution of kainate receptors bound to ancillary proteins. J Physiol 2003;547:373–385.
8.
Copits BA, Swanson GT: Dancing partners at the synapse: auxiliary subunits that shape kainate receptor function. Nat Rev Neurosci 2012;13:675–686.
9.
Laezza F, Wilding TJ, Sequeira S, Craig AM, Huettner JE: The BTB/kelch protein, KRIP6, modulates the interaction of PICK1 with GluR6 kainate receptors. Neuropharmacology 2008;55:1131–1139.
10.
Hirbec H, Francis JC, Lauri SE, et al.: Rapid and differential regulation of AMPA and kainate receptors at hippocampal mossy fibre synapses by PICK1 and GRIP. Neuron 2003;37:625–638.
11.
Piserchio A, Pellegrini M, Mehta S, et al.: The PDZ1 domain of SAP90. Characterization of structure and binding. J Biol Chem 2002;277:6967–6973.
12.
Meyer G, Varoqueaux F, Neeb A, Oschlies M, Brose N: The complexity of PDZ domain-mediated interactions at glutamatergic synapses: a case study on neuroligin. Neuropharmacology 2004;47:724–733.
13.
Beique JC, Lin DT, Kang MG, Aizawa H, Takamiya K, Huganir RL: Synapse-specific regulation of AMPA receptor function by PSD-95. Proc Natl Acad Sci U S A 2006;103:19535–19540.
14.
Ehrlich I, Klein M, Rumpel S, Malinow R: PSD-95 is required for activity-driven synapse stabilization. Proc Natl Acad Sci U S A 2007;104:4176–4181.
15.
El-Husseini AE, Schnell E, Chetkovich DM, Nicoll RA, Bredt DS: PSD-95 involvement in maturation of excitatory synapses. Science 2000;290:1364–1368.
16.
Elias GM, Funke L, Stein V, Grant SG, Bredt DS, Nicoll RA: Synapse-specific and developmentally regulated targeting of AMPA receptors by a family of MAGUK scaffolding proteins. Neuron 2006;52:307–320.
17.
de Bartolomeis A, Tomasetti C: Calcium-dependent networks in dopamine-glutamate interaction: the role of postsynaptic scaffolding proteins. Mol Neurobiol 2012;46:275–296.
18.
Gomperts SN: Clustering membrane proteins: It's all coming together with the PSD-95/SAP90 protein family. Cell 1996;84:659–662.
19.
Bats C, Groc L, Choquet D: The interaction between Stargazin and PSD-95 regulates AMPA receptor surface trafficking. Neuron 2007;53:719–734.
20.
Schnell E, Sizemore M, Karimzadegan S, Chen L, Bredt DS, Nicoll RA: Direct interactions between PSD-95 and stargazin control synaptic AMPA receptor number. Proc Natl Acad Sci U S A 2002;99:13902–13907.
21.
Migaud M, Charlesworth P, Dempster M, et al.: Enhanced long-term potentiation and impaired learning in mice with mutant postsynaptic density-95 protein. Nature 1998;396:433–439.
22.
Li B, Otsu Y, Murphy TH, Raymond LA: Developmental decrease in NMDA receptor desensitization associated with shift to synapse and interaction with postsynaptic density-95. J Neurosci 2003;23:11244–11254.
23.
Lin Y, Skeberdis VA, Francesconi A, Bennett MV, Zukin RS: Postsynaptic density protein-95 regulates NMDA channel gating and surface expression. J Neurosci 2004;24:10138–10148.
24.
Roche KW, Standley S, McCallum J, Dune Ly C, Ehlers MD, Wenthold RJ: Molecular determinants of NMDA receptor internalization. Nat Neurosci 2001;4:794–802.
25.
Wyeth MS, Pelkey KA, Petralia RS, Salter MW, McInnes RR, McBain CJ: Neto Auxiliary Protein Interactions Regulate Kainate and NMDA Receptor Subunit Localization at Mossy Fiber-CA3 Pyramidal Cell Synapses. J Neurosci 2014;34:622–628.
26.
Cousins SL, Innocent N, Stephenson FA: Neto1 associates with the NMDA receptor/amyloid precursor protein complex. J Neurochem 2013;126:554–564.
27.
Zhang W, St-Gelais F, Grabner CP, et al.: A transmembrane accessory subunit that modulates kainate-type glutamate receptors. Neuron 2009;61:385–396.
28.
Tang M, Pelkey KA, Ng D, et al.: Neto1 Is an Auxiliary Subunit of Native Synaptic Kainate Receptors. J Neurosci 2011;31:10009–10018.
29.
Fisher JL, Mott DD: Modulation of homomeric and heteromeric kainate receptors by the auxiliary subunit Neto1. J Physiol 2013;591:4711–4724.
30.
Copits BA, Robbins JS, Frausto S, Swanson GT: Synaptic targeting and functional modulation of GluK1 kainate receptors by the auxiliary neuropilin and tolloid-like (NETO) proteins. J Neurosci 2011;31:7334–7340.
31.
Straub C, Zhang W, Howe JR: Neto2 modulation of kainate receptors with different subunit compositions. J Neurosci 2011;31:8078–8082.
32.
Straub C, Hunt DL, Yamasaki M, et al.: Distinct functions of kainate receptors in the brain are determined by the auxiliary subunit Neto1. Nat Neurosci 2011;14:866–873.
33.
Griffith TN, Swanson GT: Identification of critical functional determinants of kainate receptor modulation by auxiliary protein Neto2. J Physiol 2015;593:4815–4833.
34.
Fisher JL, Mott DD: The auxiliary subunits Neto1 and Neto2 reduce voltage-dependent inhibition of recombinant kainate receptors. J Neurosci 2012;32:12928–12933.
35.
Lerma J, Paternain AV, Rodriguez-Moreno A, Lopez-Garcia JC: Molecular physiology of kainate receptors. Physiol Rev 2001;81:971–998.
36.
Lerma J, Marques JM: Kainate receptors in health and disease. Neuron 2013;80:292–311.
37.
Matute C: Therapeutic potential of kainate receptors. CNS Neurosci Ther 2011;17:661–669.
38.
Bhangoo SK, Swanson GT: Kainate receptor signaling in pain pathways. Mol Pharmacol 2013;83:307–315.
39.
Petersen KL, Iyengar S, Chappell AS, et al.: Safety, tolerability, pharmacokinetics, and effects on human experimental pain of the selective ionotropic glutamate receptor 5 (iGluR5) antagonist LY545694 in healthy volunteers. Pain 2014;155:929–936.
40.
Fisher JL: The auxiliary subunits Neto1 and Neto2 have distinct, subunit-dependent effects at recombinant GluK1- and GluK2-containing kainate receptors. Neuropharmacology 2015;99:471–480.
41.
Strutz-Seebohm N, Korniychuk G, Schwarz R, et al.: Functional significance of the kainate receptor GluR6(M836I) mutation that is linked to autism. Cell Physiol Biochem 2006;18:287–294.
42.
Mehta S, Wu H, Garner CC, Marshall J: Molecular mechanisms regulating the differential association of kainate receptor subunits with SAP90/PSD-95 and SAP97. J Biol Chem 2001;276:16092–16099.
43.
Mayer ML, Vyklicky L Jr.: Concanavalin A selectively reduces desensitization of mammalian neuronal quisqualate receptors. Proc Natl Acad Sci U S A 1989;86:1411–1415.
44.
Sommer B, Kohler M, Sprengel R, Seeburg PH: RNA editing in brain controls a determinant of ion flow in glutamate-gated channels. Cell 1991;67:11–19.
45.
Bernard A, Khrestchatisky M: Assessing the extent of RNA editing in the TMII regions of GluR5 and GluR6 kainate receptors during rat brain development. J Neurochem 1994;62:2057–2060.
46.
Barbon A, Barlati S: Glutamate receptor RNA editing in health and disease. Biochemistry (Mosc) 2011;76:882–889.
47.
Kidd FL, Isaac JT: Developmental and activity-dependent regulation of kainate receptors at thalamocortical synapses. Nature 1999;400:569–573.
48.
Vignes M, Collingridge GL: The synaptic activation of kainate receptors. Nature 1997;388:179–182.
49.
Castillo PE, Malenka RC, Nicoll RA: Kainate receptors mediate a slow postsynaptic current in hippocampal CA3 neurons. Nature 1997;388:182–186.
50.
Erreger K, Chen PE, Wyllie DJ, Traynelis SF: Glutamate receptor gating. Crit Rev Neurobiol 2004;16:187–224.
51.
Delint-Ramirez I, Fernandez E, Bayes A, Kicsi E, Komiyama NH, Grant SG: In vivo composition of NMDA receptor signaling complexes differs between membrane subdomains and is modulated by PSD-95 and PSD-93. J Neurosci 2010;30:8162–8170.
52.
Simmons RM, Li DL, Hoo KH, Deverill M, Ornstein PL, Iyengar S: Kainate GluR5 receptor subtype mediates the nociceptive response to formalin in the rat. Neuropharmacology 1998;37:25–36.
53.
Zhang WG, Zhang LC, Peng ZD, Zeng YM: Intrathecal injection of GluR6 antisense oligodeoxynucleotides alleviates acute inflammatory pain of rectum in rats. Neurosci Bull 2009;25:319–323.
54.
Grigorenko EV, Bell WL, Glazier S, Pons T, Deadwyler S: Editing status at the Q/R site of the GluR2 and GluR6 glutamate receptor subunits in the surgically excised hippocampus of patients with refractory epilepsy. Neuroreport 1998;9:2219–2224.
55.
Motazacker MM, Rost BR, Hucho T, et al.: A defect in the ionotropic glutamate receptor 6 gene (GRIK2) is associated with autosomal recessive mental retardation. Am J Hum Genet 2007;81:792–798.
56.
Alt A, Weiss B, Ogden AM, et al.: Pharmacological characterization of glutamatergic agonists and antagonists at recombinant human homomeric and heteromeric kainate receptors in vitro. Neuropharmacology 2004;46:793–806.
57.
Jackson AC, Nicoll RA: Stargazing from a new vantage—TARP modulation of AMPA receptor pharmacology. J Physiol 2011;589:5909–5910.
58.
Sheng N, Shi YS, Lomash RM, Roche KW, Nicoll RA: Neto auxiliary proteins control both the trafficking and biophysical properties of the kainate receptor GluK1. Elife 2015;4. e11682:1–19.
59.
Morimoto-Tomita M, Zhang W, Straub C, et al.: Autoinactivation of neuronal AMPA receptors via glutamate-regulated TARP interaction. Neuron 2009;61:101–112.
60.
Paternain AV, Rodriguez-Moreno A, Villarroel A, Lerma J: Activation and desensitization properties of native and recombinant kainate receptors. Neuropharmacology 1998;37:1249–1259.

Information & Authors

Information

Published In

cover image ASSAY and Drug Development Technologies
ASSAY and Drug Development Technologies
Volume 14Issue Number 2March 2016
Pages: 131 - 143
PubMed: 26991362

History

Published online: 18 March 2016
Published in print: March 2016

Permissions

Request permissions for this article.

Topics

Authors

Affiliations

Baolin Li
Neuroscience Discovery, Lilly Corporate Center, Eli Lilly and Company, Indianapolis, Indiana.
Elizabeth Rex
Neuroscience Discovery, Lilly Corporate Center, Eli Lilly and Company, Indianapolis, Indiana.
He Wang
TTx-Reagents-Proteins, Lilly Corporate Center, Eli Lilly and Company, Indianapolis, Indiana.
Yuewei Qian
TTx-Reagents-Proteins, Lilly Corporate Center, Eli Lilly and Company, Indianapolis, Indiana.
Ann Marie Ogden
Neuroscience Discovery, Lilly Corporate Center, Eli Lilly and Company, Indianapolis, Indiana.
David Bleakman
Neuroscience Discovery, Lilly Corporate Center, Eli Lilly and Company, Indianapolis, Indiana.
Kirk W. Johnson
Neuroscience Discovery, Lilly Corporate Center, Eli Lilly and Company, Indianapolis, Indiana.

Notes

Address correspondence to:Baolin Li, PhDNeuroscience DiscoveryLilly Corporate CenterEli Lilly and CompanyIndianapolis, IN 46285E-mail: [email protected]
Kirk W. Johnson, PhDNeuroscience DiscoveryLilly Corporate CenterEli Lilly and CompanyIndianapolis, IN 46285E-mail: [email protected]

Disclosure Statement

All authors are employees of Eli Lilly and Company.

Metrics & Citations

Metrics

Citations

Export citation

Select the format you want to export the citations of this publication.

View Options

Get Access

Access content

To read the fulltext, please use one of the options below to sign in or purchase access.

Society Access

If you are a member of a society that has access to this content please log in via your society website and then return to this publication.

Restore your content access

Enter your email address to restore your content access:

Note: This functionality works only for purchases done as a guest. If you already have an account, log in to access the content to which you are entitled.

View options

PDF/EPUB

View PDF/ePub

Full Text

View Full Text

Media

Figures

Other

Tables

Share

Share

Copy the content Link

Share on social media

Back to Top