1. Neuroscience

A calibrated optogenetic toolbox of stable zebrafish opsin lines

  1. Paride Antinucci
  2. Adna Dumitrescu
  3. Charlotte Deleuze  Is a corresponding author
  4. Holly J Morley
  5. Kristie Leung
  6. Tom Hagley
  7. Fumi Kubo
  8. Herwig Baier
  9. Isaac H Bianco  Is a corresponding author
  10. Claire Wyart  Is a corresponding author
  1. University College London, United Kingdom
  2. Institut du Cerveau et la Moelle épinière, Hôpital Pitié-Salpêtrière, Sorbonne Universités, UPMC Univ Paris 06, Inserm, CNRS, France
  3. Max Planck Institute of Neurobiology, Germany
https://doi.org/10.7554/eLife.54937
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Cite this article
  1. Paride Antinucci
  2. Adna Dumitrescu
  3. Charlotte Deleuze
  4. Holly J Morley
  5. Kristie Leung
  6. Tom Hagley
  7. Fumi Kubo
  8. Herwig Baier
  9. Isaac H Bianco
  10. Claire Wyart
(2020)
A calibrated optogenetic toolbox of stable zebrafish opsin lines
eLife 9:e54937.
https://doi.org/10.7554/eLife.54937
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Abstract

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.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for all Figures.

Article and author information

Author details

  1. Paride Antinucci

    Department of Neuroscience, Physiology and Pharmacology, University College London, London, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0573-5383

  2. Adna Dumitrescu

    Neurophysiology and Systems Neuroscience, Institut du Cerveau et la Moelle épinière, Hôpital Pitié-Salpêtrière, Sorbonne Universités, UPMC Univ Paris 06, Inserm, CNRS, Paris, France
    Competing interests
    No competing interests declared.

  3. Charlotte Deleuze

    Neurophysiology and Systems Neuroscience, Institut du Cerveau et la Moelle épinière, Hôpital Pitié-Salpêtrière, Sorbonne Universités, UPMC Univ Paris 06, Inserm, CNRS, Paris, France
    For correspondence
    charlotte.deleuze@icm-institute.org
    Competing interests
    No competing interests declared.

  4. Holly J Morley

    Department of Neuroscience, Physiology and Pharmacology, University College London, London, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0007-3563

  5. Kristie Leung

    Department of Neuroscience, Physiology and Pharmacology, University College London, London, United Kingdom
    Competing interests
    No competing interests declared.

  6. Tom Hagley

    Department of Neuroscience, Physiology and Pharmacology, University College London, London, United Kingdom
    Competing interests
    No competing interests declared.

  7. Fumi Kubo

    Genes, Circuits and Behavior, Max Planck Institute of Neurobiology, Martinsried, Germany
    Competing interests
    No competing interests declared.

  8. Herwig Baier

    Genes, Circuits and Behavior, Max Planck Institute of Neurobiology, Martinsried, Germany
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7268-0469

  9. Isaac H Bianco

    Department of Neuroscience, Physiology and Pharmacology, University College London, London, United Kingdom
    For correspondence
    i.bianco@ucl.ac.uk
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3149-4862

  10. Claire Wyart

    Neurophysiology & Systems neuroscience, Institut du Cerveau et la Moelle épinière, Hôpital Pitié-Salpêtrière, Sorbonne Universités, UPMC Univ Paris 06, Inserm, CNRS, Paris, France
    For correspondence
    claire.wyart@icm-institute.org
    Competing interests
    Claire Wyart, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1668-4975

Funding

Horizon 2020 Framework Programme (Marie Curie Incoming International Fellowship,H2020-MSCA-IF-2016 Project #752199)

  • Adna Dumitrescu

Human Frontier Science Program (RGP063-2018)

  • Claire Wyart

New York Stem Cell Foundation (NYSCF-R-NI39)

  • Claire Wyart

Wellcome (Sir Henry Wellcome Postdoctoral Fellowship,204708/Z/16/Z)

  • Paride Antinucci

Wellcome (Sir Henry Dale Fellowship,101195/Z/13/Z)

  • Isaac H Bianco

Royal Society (Sir Henry Dale Fellowship,101195/Z/13/Z)

  • Isaac H Bianco

University College London (Excellence Fellowship)

  • Isaac H Bianco

Human Frontier Science Program (Long-term Fellowship,LT393/2010)

  • Fumi Kubo

Deutsche Forschungsgemeinschaft (Next-Generation Optogenetics,SPP1926)

  • Herwig Baier

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Harold Burgess

Ethics

Animal experimentation: All larvae used for behavioural assays were fed Paramecia from 4 dpf onward. Animal handling and experimental procedures were approved by the UCL Animal Welfare Ethical Review Body and the UK Home Office under the Animal (Scientific Procedures) Act 1986.In vivo electrophysiological recordings were performed in 5-6 dpf zebrafish larvae from AB and Tüpfel long fin (TL) strains in accordance with the European Communities Council Directive (2010/63/EU) and French law (87/848) and approved by the Institut du Cerveau et de la Moelle épinière, the French ministry of Research and the Darwin Ethics Committee (APAFIS protocol #16469-2018071217081175v5).

Version history

  1. Received: January 7, 2020
  2. Accepted: March 27, 2020
  3. Accepted Manuscript published: March 27, 2020 (version 1)
  4. Version of Record published: April 20, 2020 (version 2)

Copyright

© 2020, Antinucci et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Paride Antinucci
  2. Adna Dumitrescu
  3. Charlotte Deleuze
  4. Holly J Morley
  5. Kristie Leung
  6. Tom Hagley
  7. Fumi Kubo
  8. Herwig Baier
  9. Isaac H Bianco
  10. Claire Wyart
(2020)
A calibrated optogenetic toolbox of stable zebrafish opsin lines
eLife 9:e54937.
https://doi.org/10.7554/eLife.54937

Share this article

https://doi.org/10.7554/eLife.54937

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