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Optogenetic strategies for high-efficiency all-optical interrogation using blue-light-sensitive opsins

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 two-photon holographic techniques requires high-sensitivity actuators to avoid photodamage and heating....

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Detalles Bibliográficos
Autores principales: Forli, Angelo, Pisoni, Matteo, Printz, Yoav, Yizhar, Ofer, Fellin, Tommaso
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8177884/
https://www.ncbi.nlm.nih.gov/pubmed/34032211
http://dx.doi.org/10.7554/eLife.63359
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author Forli, Angelo
Pisoni, Matteo
Printz, Yoav
Yizhar, Ofer
Fellin, Tommaso
author_facet Forli, Angelo
Pisoni, Matteo
Printz, Yoav
Yizhar, Ofer
Fellin, Tommaso
author_sort Forli, Angelo
collection PubMed
description 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 two-photon 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 two-photon 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.
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spelling pubmed-81778842021-06-07 Optogenetic strategies for high-efficiency all-optical interrogation using blue-light-sensitive opsins Forli, Angelo Pisoni, Matteo Printz, Yoav Yizhar, Ofer Fellin, Tommaso eLife Neuroscience 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 two-photon 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 two-photon 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. eLife Sciences Publications, Ltd 2021-05-25 /pmc/articles/PMC8177884/ /pubmed/34032211 http://dx.doi.org/10.7554/eLife.63359 Text en © 2021, Forli et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Neuroscience
Forli, Angelo
Pisoni, Matteo
Printz, Yoav
Yizhar, Ofer
Fellin, Tommaso
Optogenetic strategies for high-efficiency all-optical interrogation using blue-light-sensitive opsins
title Optogenetic strategies for high-efficiency all-optical interrogation using blue-light-sensitive opsins
title_full Optogenetic strategies for high-efficiency all-optical interrogation using blue-light-sensitive opsins
title_fullStr Optogenetic strategies for high-efficiency all-optical interrogation using blue-light-sensitive opsins
title_full_unstemmed Optogenetic strategies for high-efficiency all-optical interrogation using blue-light-sensitive opsins
title_short Optogenetic strategies for high-efficiency all-optical interrogation using blue-light-sensitive opsins
title_sort optogenetic strategies for high-efficiency all-optical interrogation using blue-light-sensitive opsins
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8177884/
https://www.ncbi.nlm.nih.gov/pubmed/34032211
http://dx.doi.org/10.7554/eLife.63359
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