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Glutamate indicators with improved activation kinetics and localization for imaging synaptic transmission
Abhi Aggarwal1,2, Rui Liu3, Yang Chen4
1Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
Nature Methods
|May 4, 2023
Summary
Researchers developed iGluSnFR3, a novel fluorescent indicator for imaging neurotransmission. This improved tool offers higher signal-to-noise ratios and better localization for precise visualization of synaptic activity in the brain.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- The fluorescent glutamate indicator iGluSnFR is crucial for studying neurotransmission.
- Existing variants have limitations including low signal-to-noise ratios and saturation.
Purpose of the Study:
- To engineer improved variants of the iGluSnFR indicator.
- To enhance signal-to-noise ratios, activation kinetics, and postsynaptic localization.
Main Methods:
- Multiassay screening in bacteria, soluble protein, and cultured neurons.
- Development of surface display constructs for nanoscopic localization.
- Simultaneous imaging and electrophysiology in mouse visual and vibrissal sensory cortex.
Main Results:
- Generated iGluSnFR3 with improved signal-to-noise ratios and kinetics.
- iGluSnFR3 demonstrates rapid, nonsaturating activation and enhanced specificity.
- Successfully imaged synaptic glutamate release and single action potentials in vivo.
- Characterized distinct patterns of sensory input in the mouse brain.
Conclusions:
- iGluSnFR3 significantly advances the ability to image synaptic activity with greater precision.
- The new indicator facilitates detailed analysis of neural circuit function.
- This work provides a powerful tool for neuroscience research.
