A calibrated optogenetic toolbox of stable zebrafish opsin lines
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- Paride Antinucci
- Department of Neuroscience, Physiology & Pharmacology, UCL, London, United Kingdom
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- Adna Dumitrescu
- Institut du Cerveau et de la Moelle épinière (ICM), Sorbonne Universités, UPMC Univ Paris 06, Inserm, CNRS, Hôpital Pitié-Salpêtrière, Paris, France
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- Charlotte Deleuze
- Institut du Cerveau et de la Moelle épinière (ICM), Sorbonne Universités, UPMC Univ Paris 06, Inserm, CNRS, Hôpital Pitié-Salpêtrière, Paris, France
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- Holly J Morley
- Department of Neuroscience, Physiology & Pharmacology, UCL, London, United Kingdom
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- Kristie Leung
- Department of Neuroscience, Physiology & Pharmacology, UCL, London, United Kingdom
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- Tom Hagley
- Department of Neuroscience, Physiology & Pharmacology, UCL, London, United Kingdom
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- Fumi Kubo
- Center for Frontier Research, National Insitute of Genetics, Mishima, Japan
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- Herwig Baier
- Department Genes – Circuits – Behavior, Max Planck Institute of Neurobiology, Martinsried, Germany
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- Isaac H Bianco
- Department of Neuroscience, Physiology & Pharmacology, UCL, London, United Kingdom
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- Claire Wyart
- Institut du Cerveau et de la Moelle épinière (ICM), Sorbonne Universités, UPMC Univ Paris 06, Inserm, CNRS, Hôpital Pitié-Salpêtrière, Paris, France
説明
<jats:p>Optogenetic 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.</jats:p>
収録刊行物
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- eLife
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eLife 9 e54937-, 2020-03-27
eLife Sciences Publications, Ltd
