Ultrafast Two-Photon Imaging of a High-Gain Voltage Indicator in Awake Behaving Mice

Authors

Vincent Villette
Mariya Chavarha
Ivan K Dimov
Jonathan Bradley
Lagnajeet Pradhan
Benjamin Mathieu
Stephen W Evans
Simon Chamberland
Dongqing Shi
Renzhi Yang
Benjamin B Kim
Annick Ayon
Abdelali Jalil
François St-Pierre
Mark J Schnitzer
Guoqiang Bi
Katalin Toth
Jun Ding
Stéphane Dieudonné
Michael Z Lin

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.