Open AccessIdentification of Fluorescent Small Molecule Compounds for Synaptic Labeling by Image-Based, High-Content Screening
Author(s) -
Matthew R. Dunn,
Umed Boltaev,
Anne Beskow,
Sergey Pampou,
Ronald Realubit,
Torcato Meira,
João VazSilva,
Rose Reeb,
Charles Karan,
Steffen Jockusch,
David Sulzer,
YoungTae Chang,
Dalibor Sameš,
Clarissa L. Waites
Publication year - 2017
Publication title -
acs chemical neuroscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.158
H-Index - 69
ISSN - 1948-7193
DOI - 10.1021/acschemneuro.7b00263
Subject(s) - colocalization , synaptic vesicle , glutamatergic , fluorescence , neurotransmission , neuroscience , biology , synapse , chemistry , biochemistry , microbiology and biotechnology , glutamate receptor , vesicle , quantum mechanics , membrane , receptor , physics
Few tools are available for noninvasive imaging of synapses in the living mammalian brain. Current paradigms require the use of genetically modified mice or viral delivery of genetic material to the brain. To develop an alternative chemical approach, utilizing the recognition of synaptic components by organic small molecules, we designed an imaging-based, high-content screen in cultured cortical neurons to identify molecules based on their colocalization with fluorescently tagged synaptic proteins. We used this approach to screen a library of ∼7000 novel fluorescent dyes, and identified a series of compounds in the xanthone family that exhibited consistent synaptic labeling. Follow-up studies with one of these compounds, CX-G3, demonstrated its ability to label acidic organelles and in particular synaptic vesicles at glutamatergic synapses in cultured neurons and murine brain tissue, indicating the potential of this screening approach to identify promising lead compounds for use as synaptic markers, sensors, and targeting devices.