Document Detail


An optimized fluorescent probe for visualizing glutamate neurotransmission.
MedLine Citation:
PMID:  23314171     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
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
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2013-01-13
Journal Detail:
Title:  Nature methods     Volume:  10     ISSN:  1548-7105     ISO Abbreviation:  Nat. Methods     Publication Date:  2013 Feb 
Date Detail:
Created Date:  2013-01-30     Completed Date:  2013-03-28     Revised Date:  2014-02-27    
Medline Journal Info:
Nlm Unique ID:  101215604     Medline TA:  Nat Methods     Country:  United States    
Other Details:
Languages:  eng     Pagination:  162-70     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Astrocytes / metabolism
Biosensing Techniques
Caenorhabditis elegans
Calcium Signaling / physiology
Escherichia coli Proteins / chemical synthesis,  diagnostic use*
Excitatory Postsynaptic Potentials / physiology
Fluorescent Dyes / chemical synthesis,  diagnostic use*,  metabolism
Glutamic Acid / metabolism*
Green Fluorescent Proteins / chemical synthesis,  diagnostic use*
Hippocampus / metabolism
Mice
Motor Cortex / metabolism
Neurons / metabolism
Photic Stimulation
Pyramidal Cells / metabolism
Recombinant Fusion Proteins / chemical synthesis,  diagnostic use*
Retina / physiology
Signal-To-Noise Ratio
Synaptic Transmission / physiology*
Zebrafish
Grant Support
ID/Acronym/Agency:
R01 EY014454/EY/NEI NIH HHS; R01 NS047325/NS/NINDS NIH HHS; T32 GM007308/GM/NIGMS NIH HHS; //Howard Hughes Medical Institute
Chemical
Reg. No./Substance:
0/Escherichia coli Proteins; 0/Fluorescent Dyes; 0/Recombinant Fusion Proteins; 0/iGluSnFR protein; 147336-22-9/Green Fluorescent Proteins; 3KX376GY7L/Glutamic Acid

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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