GluN2B Subunit Labeling with Fluorescent Probes and High-Resolution Live Imaging

Cécile Perrio; Olivier Nicole; Alain Buisson

doi:10.1007/978-1-4939-7321-7_9

GluN2B Subunit Labeling with Fluorescent Probes and High-Resolution Live Imaging

Methods Mol Biol. 2017:1677:171-183. doi: 10.1007/978-1-4939-7321-7_9.

Authors

Cécile Perrio¹, Olivier Nicole², Alain Buisson³

Affiliations

¹ Normandie University, UNICAEN, CEA, CNRS, UMR6301-ISTCT, LDM-TEP, Cyceron, Caen, France. perrio@cyceron.fr.
² Université de Bordeaux, CNRS UMR 5293, Institut des Maladies Neurodégénératives, Bordeaux, France.
³ Grenoble Institut des Neurosciences, Centre Inserm U1216, Equipe Neuropathologies et Dysfonctions Synaptiques, Université Grenoble Alpes, Grenoble, France.

PMID: 28986873
DOI: 10.1007/978-1-4939-7321-7_9

Abstract

Laser Scanning Confocal Microscopy (LSCM) imaging using an appropriate fluorescent probe enables the visualization of a molecular target with high resolution, and represents a method of choice for studying expression, subcellular location, and trafficking of receptors in living cells. The chemical, physical, and pharmacological properties of the probe remain essential. Here, we describe (1) the preparation of a specific probe for NMDAR GluN2B receptor by conjugation of fluorescein to an ifenprodil-based ligand, (2) an in vitro functional assay by calcium imaging for GluN2B binding and inhibition evaluation of the probe, and (3) the labeling and confocal imaging of GluN2B in DS-red labeled living cortical neurons.

Keywords: Calcium imaging; Confocal microscopy; Fluorescein; GluN2B; ifenprodil.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Calcium / chemistry*
Fluorescent Dyes / chemistry
Microscopy, Confocal / methods*
Piperidines / chemistry
Receptors, N-Methyl-D-Aspartate / chemistry*

Substances

Fluorescent Dyes
NR2B NMDA receptor
Piperidines
Receptors, N-Methyl-D-Aspartate
ifenprodil
Calcium