Selective dendritic localization of mRNA in Drosophila mushroom body output neurons
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 γ5β′2a 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
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
- Mani Ramaswami, Trinity College Dublin, Ireland
Publication history
- Received: September 3, 2020
- Accepted: March 15, 2021
- Accepted Manuscript published: March 16, 2021 (version 1)
- Accepted Manuscript updated: March 18, 2021 (version 2)
- 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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