Analysis of Surface Expression of NMDAR Subunits in Primary Hippocampal Neurons

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F67985823%3A_____%2F24%3A00602788" target="_blank" >RIV/67985823:_____/24:00602788 - isvavai.cz</a>

Výsledek na webu

<a href="https://doi.org/10.1007/978-1-0716-3830-9_3" target="_blank" >https://doi.org/10.1007/978-1-0716-3830-9_3</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/978-1-0716-3830-9_3" target="_blank" >10.1007/978-1-0716-3830-9_3</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Analysis of Surface Expression of NMDAR Subunits in Primary Hippocampal Neurons

Popis výsledku v původním jazyce

The expression and activity of ionotropic glutamate receptors control signal transduction at the excitatory synapses in the CNS. The NMDAR comprises two obligatory GluN1 subunits and two GluN2 or GluN3 subunits in different combinations. Each GluN subunit consists of four domains: the extracellular amino-terminal and agonist-binding domains, the transmembrane domain, and the intracellular C-terminal domain (CTD). The CTD interaction with various classes of intracellular proteins is critical for trafficking and synaptic localization of NMDARs. Amino acid mutations or the inclusion of premature stop codons in the CTD could contribute to the emergence of neurodevelopmental and neuropsychiatric disorders. Here, we describe the method of preparing primary hippocampal neurons and lentiviral particles expressing GluN subunits that can be used as a model to study cell surface expression and synaptic localization of NMDARs. We also show a simple method of fluorescence immunostaining of eGFP-tagged GluN2 subunits and subsequent microscopy technique and image analysis to study the effects of disease-associated mutations in the CTDs of GluN2A and GluN2B subunits.

Název v anglickém jazyce

Analysis of Surface Expression of NMDAR Subunits in Primary Hippocampal Neurons

Popis výsledku anglicky

The expression and activity of ionotropic glutamate receptors control signal transduction at the excitatory synapses in the CNS. The NMDAR comprises two obligatory GluN1 subunits and two GluN2 or GluN3 subunits in different combinations. Each GluN subunit consists of four domains: the extracellular amino-terminal and agonist-binding domains, the transmembrane domain, and the intracellular C-terminal domain (CTD). The CTD interaction with various classes of intracellular proteins is critical for trafficking and synaptic localization of NMDARs. Amino acid mutations or the inclusion of premature stop codons in the CTD could contribute to the emergence of neurodevelopmental and neuropsychiatric disorders. Here, we describe the method of preparing primary hippocampal neurons and lentiviral particles expressing GluN subunits that can be used as a model to study cell surface expression and synaptic localization of NMDARs. We also show a simple method of fluorescence immunostaining of eGFP-tagged GluN2 subunits and subsequent microscopy technique and image analysis to study the effects of disease-associated mutations in the CTDs of GluN2A and GluN2B subunits.

Druh

C - Kapitola v odborné knize

CEP obor

—

OECD FORD obor

30103 - Neurosciences (including psychophysiology)

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2024

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název knihy nebo sborníku

NMDA Receptors: Methods and Protocols

ISBN

978-1-0716-3829-3

Počet stran výsledku

18

Strana od-do

29-46

Počet stran knihy

292

Název nakladatele

Humana

Místo vydání

New York

Kód UT WoS kapitoly

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