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Selective Cell-Surface Expression of Triheteromeric NMDA Receptors

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NMDA Receptors

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2799))

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Abstract

NMDA-type ionotropic glutamate receptors are critically involved in many brain functions and are implicated in a variety of brain disorders. Seven NMDA receptor subunits exist (GluN1, GluN2A-D, and GluN3A-B) that assemble into tetrameric receptor subtypes with distinct functional properties and physiological roles. The majority NMDA receptors are composed of two GluN1 and two GluN2 subunits, which can assemble into four diheteromeric receptors subtypes composed of GluN1 and one type of GluN2 subunit (e.g., GluN1/2A), and presumably also six triheteromeric receptor subtypes composed of GluN1 and two different GluN2 subunits (e.g., GluN1/2A/2B). Furthermore, the GluN1 subunit exists as eight splice variants (e.g., GluN1-1a and GluN1-1b isoforms), and two different GluN1 isoforms can co-assemble to also form triheteromeric NMDA receptors (e.g., GluN1-1a/1b/2A). Here, we describe a method to faithfully express triheteromeric NMDA receptors in heterologous expression systems by controlling the identity of two of the four subunits. This method overcomes the problem that co-expression of three different NMDA receptor subunits generates two distinct diheteromeric receptor subtypes as well as one triheteromeric receptor subtype, thereby confounding studies that require a homogenous population of triheteromeric NMDA receptors. The method has been applied to selectively express recombinant triheteromeric GluN1/2A/2B, GluN1/2A/2C, GluN1/2B/2D, GluN1-1a/GluN1-1b/2A, GluN1-1a/GluN1-1b/2B receptors with negligible co-expression of the respective diheteromeric receptor subtypes. This method therefore enables quantitative evaluation of functional and pharmacological properties of triheteromeric NMDA receptors, some of which are abundant NMDA receptor subtypes in the adult brain.

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Acknowledgments

This work was supported by grants from the National Institutes of Health (R01NS116055, R01NS097536, R35NS111619, and P30GM140963).

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Authors and Affiliations

  1. Guangdong-Hong Kong-Macao Greater Bay Area Center for Brain Science and Brain-Inspired Intelligence, Southern Medical University, Guangzhou, China

    Feng Yi

  2. Department of Pharmacology, Emory University School of Medicine, Atlanta, GA, USA

    Stephen F. Traynelis

  3. Center for Structural and Functional Neuroscience, Center for Biomolecular Structure and Dynamics, Division of Biological Sciences, University of Montana, Missoula, MT, USA

    Kasper B. Hansen

Authors
  1. Feng Yi
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  2. Stephen F. Traynelis
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  3. Kasper B. Hansen
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Corresponding author

Correspondence to Kasper B. Hansen .

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Editors and Affiliations

  1. INSERM, Aix-Marseille University, Mediterranean Institute of Neurobiology (INMED), Marseille, France

    Nail Burnashev

  2. INSERM, Aix-Marseille University, Mediterranean Institute of Neurobiology (INMED), Marseille, France

    Pierre Szepetowski

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Yi, F., Traynelis, S.F., Hansen, K.B. (2024). Selective Cell-Surface Expression of Triheteromeric NMDA Receptors. In: Burnashev, N., Szepetowski, P. (eds) NMDA Receptors. Methods in Molecular Biology, vol 2799. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3830-9_5

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  • DOI: https://doi.org/10.1007/978-1-0716-3830-9_5

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3829-3

  • Online ISBN: 978-1-0716-3830-9

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