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

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

doi:10.1016/j.cell.2019.11.004

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

Cell. 2019 Dec 12;179(7):1590-1608.e23. doi: 10.1016/j.cell.2019.11.004.

Authors

Vincent Villette¹, Mariya Chavarha², Ivan K Dimov³, Jonathan Bradley¹, Lagnajeet Pradhan³, Benjamin Mathieu¹, Stephen W Evans⁴, Simon Chamberland⁵, Dongqing Shi⁶, Renzhi Yang⁷, Benjamin B Kim⁸, Annick Ayon¹, Abdelali Jalil⁹, François St-Pierre¹⁰, Mark J Schnitzer¹¹, Guoqiang Bi¹², Katalin Toth⁵, Jun Ding¹³, Stéphane Dieudonné¹⁴, Michael Z Lin¹⁵

Affiliations

¹ Institut de Biologie de l'École Normale Supérieure (IBENS), École Normale Supérieure, CNRS, INSERM, PSL Research University, Paris 75005, France.
² Department of Neurobiology, Stanford University, Stanford, CA 94305, USA; Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.
³ Department of Neurobiology, Stanford University, Stanford, CA 94305, USA; CNC Program, Stanford University, Stanford, CA 94305, USA.
⁴ Department of Neurobiology, Stanford University, Stanford, CA 94305, USA.
⁵ Department of Psychiatry and Neuroscience, CERVO Brain Research Centre, Université Laval, Quebec City, QC G1J 2G3, Canada.
⁶ Department of Neurobiology, Stanford University, Stanford, CA 94305, USA; School of Life Sciences, University of Science and Technology of China, Hefei 230026, China.
⁷ Department of Neurosurgery, Stanford University, Stanford, CA 94305, USA; Biology PhD Program, Stanford University, Stanford, CA 94305, USA.
⁸ Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.
⁹ Université de Paris, SPPIN - Saints-Pères Paris Institute for the Neurosciences, CNRS, Paris F-75006, France.
¹⁰ Department of Neuroscience, Baylor College of Medicine, Houston, TX 77030, USA.
¹¹ CNC Program, Stanford University, Stanford, CA 94305, USA; Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA.
¹² School of Life Sciences, University of Science and Technology of China, Hefei 230026, China; CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai 20031, China.
¹³ Department of Neurosurgery, Stanford University, Stanford, CA 94305, USA.
¹⁴ Institut de Biologie de l'École Normale Supérieure (IBENS), École Normale Supérieure, CNRS, INSERM, PSL Research University, Paris 75005, France. Electronic address: dieudon@biologie.ens.fr.
¹⁵ Department of Neurobiology, Stanford University, Stanford, CA 94305, USA; Department of Bioengineering, Stanford University, Stanford, CA 94305, USA. Electronic address: mzlin@stanford.edu.

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.

Keywords: ASAP3; GEVI; RAMP; ULoVE; action potential; electrophysiology; electroporation; theta oscillation; two-photon; voltage imaging.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Action Potentials
Animals
Brain / metabolism
Brain / physiology*
CHO Cells
Cells, Cultured
Cricetinae
Cricetulus
Female
GTPase-Activating Proteins / genetics*
GTPase-Activating Proteins / metabolism
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism
HEK293 Cells
Humans
Male
Mice
Mice, Inbred C57BL
Microscopy, Fluorescence, Multiphoton / methods*
Optogenetics / methods*
Phosphoric Monoester Hydrolases / genetics
Phosphoric Monoester Hydrolases / metabolism
Rats
Rats, Sprague-Dawley
Running
Theta Rhythm*
Wakefulness*

Substances

GTPase-Activating Proteins
Green Fluorescent Proteins
Phosphoric Monoester Hydrolases

Abstract

Publication types

MeSH terms

Substances

Grants and funding