Selective dendritic localization of mRNA in Drosophila mushroom body output neurons

  1. Jessica Mitchell
  2. Carlas S Smith
  3. Josh Titlow
  4. Nils Otto
  5. Pieter van Velde
  6. Martin J Booth
  7. Ilan Davis
  8. Scott Waddell  Is a corresponding author
  1. University of Oxford, United Kingdom
  2. Delft University of Technology, Netherlands

Abstract

Memory-relevant neuronal plasticity is believed to require local translation of new proteins at synapses. Understanding this process requires the visualization of the relevant mRNAs within these neuronal compartments. Here we used single-molecule fluorescence in situ hybridization (smFISH) to localize mRNAs at subcellular resolution in the adult Drosophila brain. mRNAs for subunits of nicotinic acetylcholine receptors and kinases could be detected within the dendrites of co-labelled Mushroom Body Output Neurons (MBONs) and their relative abundance showed cell-specificity. Moreover, aversive olfactory learning produced a transient increase in the level of CaMKII mRNA within the dendritic compartments of the 52a MBONs. Localization of specific mRNAs in MBONs before and after learning represents a critical step towards deciphering the role of dendritic translation in the neuronal plasticity underlying behavioural change in Drosophila.

Data availability

Pipeline code and the User Manual are available in the GitHub repository at [https://github.com/qnano/smFISHlearning].An example dataset of raw and processed images is available at [https://figshare.com/articles/dataset/Example_data/13568438].All other processed and raw datasets that support the findings of this study are available at [https://doi.org/10.6084/m9.figshare.13573475].

Article and author information

Author details

  1. Jessica Mitchell

    Centre for Neural Circuits & Behaviour, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  2. Carlas S Smith

    Centre for Neural Circuits & Behaviour, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  3. Josh Titlow

    Department of Biochemistry, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  4. Nils Otto

    Centre for Neural Circuits & Behaviour, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9713-4088
  5. Pieter van Velde

    Delft Center for Systems and Control, Delft University of Technology, Kantens, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7281-8026
  6. Martin J Booth

    Department of Engineering, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  7. Ilan Davis

    Department of Biochemistry, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5385-3053
  8. Scott Waddell

    Centre for Neural Circuits & Behaviour, University of Oxford, Oxford, United Kingdom
    For correspondence
    scott.waddell@cncb.ox.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4503-6229

Funding

Wellcome Trust (200846/Z/16/Z)

  • Scott Waddell

Wellcome Trust (203261/Z/16/Z)

  • Scott Waddell

ERC

  • Scott Waddell

Netherlands Organisation for Scientific Research

  • Carlas S Smith

Wellcome Trust (107457)

  • Ilan Davis

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

Reviewing Editor

  1. Mani Ramaswami, Trinity College Dublin, Ireland

Publication history

  1. Received: September 3, 2020
  2. Accepted: March 15, 2021
  3. Accepted Manuscript published: March 16, 2021 (version 1)
  4. Accepted Manuscript updated: March 18, 2021 (version 2)
  5. Version of Record published: March 26, 2021 (version 3)

Copyright

© 2021, Mitchell 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. Jessica Mitchell
  2. Carlas S Smith
  3. Josh Titlow
  4. Nils Otto
  5. Pieter van Velde
  6. Martin J Booth
  7. Ilan Davis
  8. Scott Waddell
(2021)
Selective dendritic localization of mRNA in Drosophila mushroom body output neurons
eLife 10:e62770.
https://doi.org/10.7554/eLife.62770

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