High-throughput automated methods for classical and operant conditioning of Drosophila larvae

  1. Elise C Croteau-Chonka
  2. Michael S Clayton
  3. Lalanti Venkatasubramanian
  4. Samuel N Harris
  5. Benjamin M W Jones
  6. Lakshmi Narayan
  7. Michael Winding
  8. Jean-Baptiste Masson
  9. Marta Zlatic  Is a corresponding author
  10. Kristina T Klein  Is a corresponding author
  1. University of Cambridge, United Kingdom
  2. MRC Laboratory of Molecular Biology, United Kingdom
  3. Janelia Research Campus, United States
  4. Institut Pasteur, France

Abstract

Learning which stimuli (classical conditioning) or which actions (operant conditioning) predict rewards or punishments can improve chances of survival. However, the circuit mechanisms that underlie distinct types of associative learning are still not fully understood. Automated, high-throughput paradigms for studying different types of associative learning, combined with manipulation of specific neurons in freely behaving animals, can help advance this field. The Drosophila melanogaster larva is a tractable model system for studying the circuit basis of behaviour, but many forms of associative learning have not yet been demonstrated in this animal. Here, we developed a high-throughput (i. e. multi-larva) training system that combines real-time behaviour detection of freely moving larvae with targeted opto- and thermogenetic stimulation of tracked animals. Both stimuli are controlled in either open- or closed-loop, and delivered with high temporal and spatial precision. Using this tracker, we show for the first time that Drosophila larvae can perform classical conditioning with no overlap between sensory stimuli (i. e. trace conditioning). We also demonstrate that larvae are capable of operant conditioning by inducing a bend direction preference through optogenetic activation of reward-encoding serotonergic neurons. Our results extend the known associative learning capacities of Drosophila larvae. Our automated training rig will facilitate the study of many different forms of associative learning and the identification of the neural circuits that underpin them.

Data availability

All data used to generate figures 2-5, as well as all supplemental figures, are now submitted as source data files. We also now submit CAD drawings for the multi-larva tracker.

Article and author information

Author details

  1. Elise C Croteau-Chonka

    Department of Zoology, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5116-3772
  2. Michael S Clayton

    MRC Laboratory of Molecular Biology, Cambridge, United Kingdom
    Competing interests
    No competing interests declared.
  3. Lalanti Venkatasubramanian

    Department of Zoology, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    No competing interests declared.
  4. Samuel N Harris

    MRC Laboratory of Molecular Biology, Cambridge, United Kingdom
    Competing interests
    No competing interests declared.
  5. Benjamin M W Jones

    MRC Laboratory of Molecular Biology, Cambridge, United Kingdom
    Competing interests
    No competing interests declared.
  6. Lakshmi Narayan

    Janelia Research Campus, Ashburn, United States
    Competing interests
    No competing interests declared.
  7. Michael Winding

    Department of Zoology, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    No competing interests declared.
  8. Jean-Baptiste Masson

    Department of Computational Biology and Neuroscience, Institut Pasteur, Paris, France
    Competing interests
    No competing interests declared.
  9. Marta Zlatic

    MRC Laboratory of Molecular Biology, Cambridge, United Kingdom
    For correspondence
    mzlatic@mrc-lmb.cam.ac.uk
    Competing interests
    Marta Zlatic, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3149-2250
  10. Kristina T Klein

    Department of Zoology, University of Cambridge, Cambridge, United Kingdom
    For correspondence
    kristina.t.klein@gmail.com
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8772-3628

Funding

Gates Cambridge Trust

  • Marta Zlatic

Cambridge Trust

  • Marta Zlatic

HHMI Janelia Visiting Scientist Program

  • Marta Zlatic

University of Cambridge, Trinity College

  • Marta Zlatic

HHMI Janelia

  • Marta Zlatic

European Research Council

  • Marta Zlatic

Wellcome Trust

  • Marta Zlatic

Medical Research Council

  • Marta Zlatic

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

Copyright

© 2022, Croteau-Chonka 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.

Metrics

  • 1,823
    views
  • 238
    downloads
  • 9
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Elise C Croteau-Chonka
  2. Michael S Clayton
  3. Lalanti Venkatasubramanian
  4. Samuel N Harris
  5. Benjamin M W Jones
  6. Lakshmi Narayan
  7. Michael Winding
  8. Jean-Baptiste Masson
  9. Marta Zlatic
  10. Kristina T Klein
(2022)
High-throughput automated methods for classical and operant conditioning of Drosophila larvae
eLife 11:e70015.
https://doi.org/10.7554/eLife.70015

Share this article

https://doi.org/10.7554/eLife.70015

Further reading

    1. Computational and Systems Biology
    2. Microbiology and Infectious Disease
    Ritwik Maity, Xuepei Zhang ... Javier Sancho
    Research Article

    Antimicrobial resistance is responsible for an alarming number of deaths, estimated at 5 million per year. To combat priority pathogens, like Helicobacter pylori, the development of novel therapies is of utmost importance. Understanding the molecular alterations induced by medications is critical for the design of multi-targeting treatments capable of eradicating the infection and mitigating its pathogenicity. However, the application of bulk omics approaches for unraveling drug molecular mechanisms of action is limited by their inability to discriminate between target-specific modifications and off-target effects. This study introduces a multi-omics method to overcome the existing limitation. For the first time, the Proteome Integral Solubility Alteration (PISA) assay is utilized in bacteria in the PISA-Express format to link proteome solubility with different and potentially immediate responses to drug treatment, enabling us the resolution to understand target-specific modifications and off-target effects. This study introduces a comprehensive method for understanding drug mechanisms and optimizing the development of multi-targeting antimicrobial therapies.

    1. Computational and Systems Biology
    Harlan P Stevens, Carly V Winegar ... Stephen R Piccolo
    Research Article

    To help maximize the impact of scientific journal articles, authors must ensure that article figures are accessible to people with color-vision deficiencies (CVDs), which affect up to 8% of males and 0.5% of females. We evaluated images published in biology- and medicine-oriented research articles between 2012 and 2022. Most included at least one color contrast that could be problematic for people with deuteranopia (‘deuteranopes’), the most common form of CVD. However, spatial distances and within-image labels frequently mitigated potential problems. Initially, we reviewed 4964 images from eLife, comparing each against a simulated version that approximated how it might appear to deuteranopes. We identified 636 (12.8%) images that we determined would be difficult for deuteranopes to interpret. Our findings suggest that the frequency of this problem has decreased over time and that articles from cell-oriented disciplines were most often problematic. We used machine learning to automate the identification of problematic images. For a hold-out test set from eLife (n=879), a convolutional neural network classified the images with an area under the precision-recall curve of 0.75. The same network classified images from PubMed Central (n=1191) with an area under the precision-recall curve of 0.39. We created a Web application (https://bioapps.byu.edu/colorblind_image_tester); users can upload images, view simulated versions, and obtain predictions. Our findings shed new light on the frequency and nature of scientific images that may be problematic for deuteranopes and motivate additional efforts to increase accessibility.