Glutamate indicators with improved activation kinetics and localization for imaging synaptic transmission.

Autor: Aggarwal A; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.; Allen Institute for Neural Dynamics, Seattle, WA, USA., Liu R; Department of Physics, University of California, San Diego, La Jolla, CA, USA., Chen Y; Institute of Neuroscience and Cluster for Systems Neurology (SyNergy), Technical University of Munich (TUM), Munich, Germany., Ralowicz AJ; Department of Biological Sciences, Dartmouth College, Hanover, NH, USA., Bergerson SJ; Department of Biological Sciences, Dartmouth College, Hanover, NH, USA., Tomaska F; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.; Department of Physiology, Second Faculty of Medicine, Charles University, Prague, Czech Republic., Mohar B; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA., Hanson TL; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA., Hasseman JP; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA., Reep D; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA., Tsegaye G; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA., Yao P; Neurosciences Graduate Program, University of California San Diego, La Jolla, CA, USA., Ji X; Department of Physics, University of California, San Diego, La Jolla, CA, USA., Kloos M; Institute of Neuroscience and Cluster for Systems Neurology (SyNergy), Technical University of Munich (TUM), Munich, Germany., Walpita D; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA., Patel R; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA., Mohr MA; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.; Department of Biosystems Science and Engineering, Swiss Federal Institute of Technology (ETH) Zurich, Basel, Switzerland., Tillberg PW; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA., Looger LL; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.; Howard Hughes Medical Institute, Department of Neurosciences, University of California, San Diego, La Jolla, CA, USA., Marvin JS; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA., Hoppa MB; Department of Biological Sciences, Dartmouth College, Hanover, NH, USA., Konnerth A; Institute of Neuroscience and Cluster for Systems Neurology (SyNergy), Technical University of Munich (TUM), Munich, Germany., Kleinfeld D; Department of Physics, University of California, San Diego, La Jolla, CA, USA.; Section of Neurobiology, University of California, San Diego, La Jolla, CA, USA., Schreiter ER; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA., Podgorski K; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA. kaspar.podgorski@alleninstitute.org.; Allen Institute for Neural Dynamics, Seattle, WA, USA. kaspar.podgorski@alleninstitute.org.
Jazyk: angličtina
Zdroj: Nature methods [Nat Methods] 2023 Jun; Vol. 20 (6), pp. 925-934. Date of Electronic Publication: 2023 May 04.
DOI: 10.1038/s41592-023-01863-6
Abstrakt: The fluorescent glutamate indicator iGluSnFR enables imaging of neurotransmission with genetic and molecular specificity. However, existing iGluSnFR variants exhibit low in vivo signal-to-noise ratios, saturating activation kinetics and exclusion from postsynaptic densities. Using a multiassay screen in bacteria, soluble protein and cultured neurons, we generated variants with improved signal-to-noise ratios and kinetics. We developed surface display constructs that improve iGluSnFR's nanoscopic localization to postsynapses. The resulting indicator iGluSnFR3 exhibits rapid nonsaturating activation kinetics and reports synaptic glutamate release with decreased saturation and increased specificity versus extrasynaptic signals in cultured neurons. Simultaneous imaging and electrophysiology at individual boutons in mouse visual cortex showed that iGluSnFR3 transients report single action potentials with high specificity. In vibrissal sensory cortex layer 4, we used iGluSnFR3 to characterize distinct patterns of touch-evoked feedforward input from thalamocortical boutons and both feedforward and recurrent input onto L4 cortical neuron dendritic spines.
(© 2023. The Author(s).)
Databáze: MEDLINE
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