A calibrated optogenetic toolbox of stable zebrafish opsin lines

eLife

Fumi Kubo¹  Herwig Baier²  Isaac H Bianco³  Paride Antinucci³  Holly J Morley³  Tom Hagley³  Kristie Leung³  Claire Wyart⁴  Adna Dumitrescu⁴  Charlotte Deleuze⁴ 

[1] Center for Frontier Research, National Insitute of Genetics, Mishima, Japan;Department Genes – Circuits – Behavior, Max Planck Institute of Neurobiology, Martinsried, Germany;Department Genes – Circuits – Behavior, Max Planck Institute of Neurobiology, Martinsried, Germany;Department of Neuroscience, Physiology & Pharmacology, UCL, London, United Kingdom;Institut du Cerveau et de la Moelle épinière (ICM), Sorbonne Universités, UPMC Univ Paris 06, Inserm, CNRS, Hôpital Pitié-Salpêtrière, Paris, France;

关键词: optogenetics;    electrophysiology;    transgenesis;    CoChR;    ChrimsonR;    GtACR;    Zebrafish;   

DOI  :  10.7554/eLife.54937

来源: publisher

【 摘 要 】

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.

【 授权许可】

CC BY