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Visualization of Glutamatergic Neurotransmission in Diverse Model Organisms with Genetically Encoded Indicators

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New Technologies for Glutamate Interaction: Neurons and Glia. ;2780:3–34. doi: 10.1007/978-1-0716-3742-5_1
Visualization of Glutamatergic Neurotransmission in Diverse Model Organisms with Genetically Encoded Indicators GENIETool Translation Team (T3)
Aggarwal A, Chan J, Waring AK, Negrean A, Marvin JS, Podgorski K, Looger LL, Kukley M
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Abstract

Glutamate is the principal excitatory neurotransmitter, and occasionally subserves inhibitory roles, in the vertebrate nervous system. Glutamatergic synapses are dense in the vertebrate brain, at \textasciitilde1/μm3. Glutamate is released from and onto diverse components of the nervous system, including neurons, glia, and other cells. Methods for glutamate detection are critically important for understanding the function of synapses and neural circuits in normal physiology, development, and disease. Here we describe the development, optimization, and deployment of genetically encoded fluorescent glutamate indicators. We review the theoretical considerations governing glutamate sensor properties from first principles of synapse biology, microscopy, and protein structure-function relationships. We provide case studies of the state-of-the-art iGluSnFR glutamate sensor, encompassing design and optimization, mechanism of action, in vivo imaging, data analysis, and future directions. We include detailed protocols for iGluSnFR imaging in common preparations (bacteria, cell culture, and brain slices) and model organisms (worm, fly, fish, rodent).

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Janelia Authors

Abhi Aggarwal
Amelia Waring
Jonathan Marvin
Kaspar Podgorski
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