Hyperspectral multiplex single-particle tracking of different receptor subtypes labeled with quantum dots in live neurons.
The efficacy of existing therapies and the discovery of innovative treatments for central nervous system (CNS) diseases have been limited by the lack of appropriate methods to investigate complex molecular processes at the synaptic level. To improve our capability to investigate complex mechanisms of synaptic signaling and remodeling, we designed a fluorescence hyperspectral imaging platform to simultaneously track different subtypes of individual neurotransmitter receptors trafficking in and out of synapses. This imaging platform allows simultaneous image acquisition of at least five fluorescent markers in living neurons with a high-spatial resolution. We used quantum dots emitting at different wavelengths and functionalized to specifically bind to single receptors on the membrane of living neurons. The hyperspectral imaging platform enabled the simultaneous optical tracking of five different synaptic proteins, including subtypes of glutamate receptors (mGluR and AMPAR) and postsynaptic signaling proteins. It also permitted the quantification of their mobility after treatments with various pharmacological agents. This technique provides an efficient method to monitor several synaptic proteins at the same time, which could accelerate the screening of effective compounds for treatment of CNS disorders.
Institut Universitaire en santé mentale de Québec, 2601 Chemin de la Canardière, Québec G1J 2G3, Canada.
Simon
S
Labrecque
Photon Etc., 5795 de Gaspé ave., Montréal, Québec H2S 2X3, Canada.
Jean-Philippe
JP
Sylvestre
Photon Etc., 5795 de Gaspé ave., Montréal, Québec H2S 2X3, Canada.
Stephane
S
Marcet
Photon Etc., 5795 de Gaspé ave., Montréal, Québec H2S 2X3, Canada.
Francesca
F
Mangiarini
Photon Etc., 5795 de Gaspé ave., Montréal, Québec H2S 2X3, Canada.
Brice
B
Bourgoin
Photon Etc., 5795 de Gaspé ave., Montréal, Québec H2S 2X3, Canada.
Marc
M
Verhaegen
Photon Etc., 5795 de Gaspé ave., Montréal, Québec H2S 2X3, Canada.
Sébastien
S
Blais-Ouellette
Institut Universitaire en santé mentale de Québec, 2601 Chemin de la Canardière, Québec G1J 2G3, CanadacUniversité Laval, Department of Biochemistry, Microbiology, and Bio-informatics, Québec G1K 7P4, Canada.
Paul
P
De Koninck
D000818:Animals; D004867:Equipment Design; D005456:Fluorescent Dyes / Q000737:chemistry*; D006624:Hippocampus / Q000166:cytology / Q000000981:diagnostic imaging; D057054:Molecular Imaging / Q000379:methods*; D009474:Neurons / Q000166:cytology*; D061848:Optical Imaging / Q000379:methods*; D045663:Quantum Dots / Q000737:chemistry*; D051381:Rats
D005456:Fluorescent Dyes