RT Journal Article
SR Electronic
T1 A calibrated optogenetic toolbox of stable zebrafish opsin lines
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.01.13.904185
DO 10.1101/2020.01.13.904185
A1 P Antinucci
A1 AS Dumitrescu
A1 C Deleuze
A1 HJ Morley
A1 K Leung
A1 T Hagley
A1 F Kubo
A1 H Baier
A1 IH Bianco
A1 C Wyart
YR 2020
UL http://biorxiv.org/content/early/2020/01/15/2020.01.13.904185.abstract
AB 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.