An optimized fluorescent probe for visualizing glutamate neurotransmission View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2013-02

AUTHORS

Jonathan S Marvin, Bart G Borghuis, Lin Tian, Joseph Cichon, Mark T Harnett, Jasper Akerboom, Andrew Gordus, Sabine L Renninger, Tsai-Wen Chen, Cornelia I Bargmann, Michael B Orger, Eric R Schreiter, Jonathan B Demb, Wen-Biao Gan, S Andrew Hires, Loren L Looger

ABSTRACT

We describe an intensity-based glutamate-sensing fluorescent reporter (iGluSnFR) with signal-to-noise ratio and kinetics appropriate for in vivo imaging. We engineered iGluSnFR in vitro to maximize its fluorescence change, and we validated its utility for visualizing glutamate release by neurons and astrocytes in increasingly intact neurological systems. In hippocampal culture, iGluSnFR detected single field stimulus-evoked glutamate release events. In pyramidal neurons in acute brain slices, glutamate uncaging at single spines showed that iGluSnFR responds robustly and specifically to glutamate in situ, and responses correlate with voltage changes. In mouse retina, iGluSnFR-expressing neurons showed intact light-evoked excitatory currents, and the sensor revealed tonic glutamate signaling in response to light stimuli. In worms, glutamate signals preceded and predicted postsynaptic calcium transients. In zebrafish, iGluSnFR revealed spatial organization of direction-selective synaptic activity in the optic tectum. Finally, in mouse forelimb motor cortex, iGluSnFR expression in layer V pyramidal neurons revealed task-dependent single-spine activity during running. More... »

PAGES

162

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nmeth.2333

DOI

http://dx.doi.org/10.1038/nmeth.2333

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1032394242

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/23314171


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