Publication Date
12-12-2019
Journal
Cell
DOI
10.1016/j.cell.2019.11.004
PMID
31835034
PMCID
PMC6941988
PubMedCentral® Posted Date
12-12-2020
PubMedCentral® Full Text Version
Author MSS
Published Open-Access
yes
Keywords
Action Potentials, Animals, Brain, CHO Cells, Cells, Cultured, Cricetinae, Cricetulus, Female, GTPase-Activating Proteins, Green Fluorescent Proteins, HEK293 Cells, Humans, Male, Mice, Mice, Inbred C57BL, Microscopy, Fluorescence, Multiphoton, Optogenetics, Phosphoric Monoester Hydrolases, Rats, Rats, Sprague-Dawley, Running, Theta Rhythm, Wakefulness
Abstract
Optical interrogation of voltage in deep brain locations with cellular resolution would be immensely useful for understanding how neuronal circuits process information. Here, we report ASAP3, a genetically encoded voltage indicator with 51% fluorescence modulation by physiological voltages, submillisecond activation kinetics, and full responsivity under two-photon excitation. We also introduce an ultrafast local volume excitation (ULoVE) method for kilohertz-rate two-photon sampling in vivo with increased stability and sensitivity. Combining a soma-targeted ASAP3 variant and ULoVE, we show single-trial tracking of spikes and subthreshold events for minutes in deep locations, with subcellular resolution and with repeated sampling over days. In the visual cortex, we use soma-targeted ASAP3 to illustrate cell-type-dependent subthreshold modulation by locomotion. Thus, ASAP3 and ULoVE enable high-speed optical recording of electrical activity in genetically defined neurons at deep locations during awake behavior.
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Biochemistry, Biophysics, and Structural Biology Commons, Biology Commons, Cognitive Neuroscience Commons, Cognitive Science Commons, Neurology Commons, Neurosciences Commons
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