Optogenetic strategies for high-efficiency all-optical interrogation using blue light-sensitive opsins
- Italian Institute of Technology, Italy
- Istituto Italiano di Tecnologia, Italy
- Weizmann Institute of Science, Israel
Abstract
All-optical methods for imaging and manipulating brain networks with high spatial resolution are fundamental to study how neuronal ensembles drive behavior. Stimulation of neuronal ensembles using holographic techniques requires high-sensitivity actuators to avoid photodamage and heating. Moreover, two-photon-excitable opsins should be insensitive to light at wavelengths used for imaging. To achieve this goal, we developed a novel soma-targeted variant of the large-conductance blue light-sensitive opsin CoChR (stCoChR). In the mouse cortex in vivo, we combined holographic two-photon stimulation of stCoChR with an amplified laser tuned at the opsin absorption peak and imaging of the red-shifted indicator jRCaMP1a. Compared to previously characterized blue light-sensitive soma-targeted opsins in vivo, stCoChR allowed neuronal stimulation with more than 10-fold lower average power and no spectral crosstalk. The combination of stCoChR, tuned amplified laser stimulation, and red-shifted functional indicators promises to be a powerful tool for large-scale interrogation of neural networks in the intact brain.
Data availability
We provide Source Data for the data plotted in all figures as Excel Source Data files.
Article and author information
Author details
Funding
H2020 European Research Council (647725)
- Tommaso Fellin
Candice Appleton Family Trust
- Ofer Yizhar
H2020-RIA (DEEPER 101016787)
- Ofer Yizhar
NIH Brain Initiative (U01 NS090576)
- Tommaso Fellin
NIH Brain Initiative (U19 NS107464)
- Tommaso Fellin
H2020-RIA (DEEPER 101016787)
- Tommaso Fellin
H2020 European Research Council (819496)
- Ofer Yizhar
Human Frontier Science Program
- Ofer Yizhar
Brain and Behavior Research Foundation
- Ofer Yizhar
Ilse Katz Institute for Material Sciences and Magnetic Resonance Research
- Ofer Yizhar
Adelis Prize for Brain Research
- Ofer Yizhar
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: All experiments involving animals were approved by the IIT Animal Welfare Body, by theNational Council on Animal Care of the Italian Ministry of Health (authorization #34/2015-PR, #1084/2020-PR), and by the Institutional Animal Care and Use Committee at the Weizmann Institute of Science, and carried out in accordance with the guidelines established by the European Communities Council Directive.
Copyright
© 2021, Forli et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Optogenetic strategies for high-efficiency all-optical interrogation using blue light-sensitive opsins
eLife 10:e63359.
https://doi.org/10.7554/eLife.63359
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