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TitleA calibrated optogenetic toolbox of stable zebrafish opsin lines
AuthorParide Antinucci 1†, Adna Dumitrescu 2†, Charlotte Deleuze 2 , Holly J Morley 1,Kristie Leung 1, Tom Hagley 1, Fumi Kubo 3,4, Herwig Baier 4, Isaac H Bianco 1‡*, Claire Wyart 2‡*
Affiliation(s)1Department of Neuroscience, Physiology & Pharmacology, UCL, London, United Kingdom; 2 Institut du Cerveau et de la Moelle e´ pinie` re (ICM), Sorbonne Universite´ s, UPMC Univ Paris 06, Inserm, CNRS, Hoˆ pital Pitie´ -Salpeˆ trie` re, Paris, France; 3 Center for Frontier Research, National Insitute of Genetics, Mishima, Japan; 4 Department Genes – Circuits – Behavior, Max Planck Institute of Neurobiology, Martinsried, Germany
Published eLife 2020;9:e54937 DOI: https://doi.org/10.7554/eLife.54937
Keywordoptogenetic toolbox
SnippetOptogenetic actuators with diverse spectral tuning, ion selectivity and kinetics are constantly being engineered providing powerful tools for controlling neural activity with subcellular resolution and millisecond precision.
AbstractOptogenetic actuators with diverse spectral tuning, ion selectivity and kinetics are constantly being engineered providing powerful tools for controlling neural activity with subcellular resolution and millisecond precision. Achieving reliable and interpretable in vivo optogenetic manipulations requires reproducible actuator expression and calibration of photocurrents in target neurons. Here, we developed nine transgenic zebrafish lines for stable opsin expression and calibrated their efficacy in vivo. We first used high-throughput behavioural assays to compare opsin ability to elicit or silence neural activity. Next, we performed in vivo whole-cell electrophysiological recordings to quantify the amplitude and kinetics of photocurrents and test opsin ability to precisely control spiking. We observed substantial variation in efficacy, associated with differences in both opsin expression level and photocurrent characteristics, and identified conditions for optimal use of the most efficient opsins. Overall, our calibrated optogenetic toolkit will facilitate the design of controlled optogenetic circuit manipulations.

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