Live imaging of axonal transport in Drosophila pupal brain explants

Nature Protocols
Caroline MedioniFlorence Besse

Abstract

Axonal transport is essential for the initial growth, maintenance and synaptic plasticity of axons, and altered axonal transport has been observed in different models of neurodegenerative pathologies. Dissecting the mechanisms underlying axonal transport in developing or degenerating brains requires dynamic imaging of axonal cargo movement in living samples. Whereas methods exist to image axonal transport in Drosophila larval neurons, they are not suitable to follow this process during metamorphosis, when brains undergo extensive remodeling. Here we present a simple method that enables confocal imaging of both fast and slow axonal transport in Drosophila pupal brain explants. We describe how to prepare chambers adapted for live imaging, how to maintain brain explants under physiological conditions and how to monitor and quantitatively analyze the movement of fluorescently labeled cargoes. This protocol requires minimal equipment and is ideally suited for experiments that combine genetics, optogenetics and pharmacological approaches. The brains can be prepared for image acquisition in 1.5 h, and the protocol can be performed easily in any fly laboratory.

References

Feb 9, 2002·Brain Research. Molecular Brain Research·E E MorrisonJ M Askham
Sep 27, 2002·Genesis : the Journal of Genetics and Development·Joseph B Duffy
Sep 27, 2002·Genesis : the Journal of Genetics and Development·Yong Q ZhangKendal Broadie
Feb 16, 2005·Nature Reviews. Neuroscience·Nobutaka Hirokawa, Reiko Takemura
Feb 10, 2006·Molecular Biology of the Cell·Aaron D PillingWilliam M Saxton
Aug 25, 2006·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Robert KraftLinda L Restifo
Jun 6, 2008·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Derya AyazBassem A Hassan
Jun 19, 2008·Annual Review of Neuroscience·Kurt J De VosChristopher C J Miller
Jul 16, 2008·Proceedings of the National Academy of Sciences of the United States of America·Barret D PfeifferGerald M Rubin
Jun 9, 2009·Proceedings of the National Academy of Sciences of the United States of America·Liria M Masuda-NakagawaReinhard F Stocker
Jun 8, 2010·Nature Methods·Johannes D SeeligVivek Jayaraman
Aug 11, 2010·Genetics·Barret D PfeifferGerald M Rubin
Jan 31, 2013·Nature Reviews. Neuroscience·Stéphanie Millecamps, Jean-Pierre Julien
Apr 13, 2013·Fly·Lyubov Veverytsa, Douglas W Allan
Apr 20, 2013·Nature Protocols·Boris EggerSimon G Sprecher
Sep 13, 2013·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Xin XuElizabeth R Gavis
Sep 14, 2013·Journal of Neurochemistry·Sarah LenzAaron Voigt
Mar 25, 2014·Current Biology : CB·Caroline MedioniFlorence Besse
May 6, 2014·Current Opinion in Neurobiology·Fengwei Yu, Oren Schuldiner
Oct 1, 1997·Development Genes and Evolution·Marco TettamantiH Reichert

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Citations

Oct 26, 2016·Brain Research Bulletin·André VoelzmannAndreas Prokop
Jun 15, 2019·Nature Communications·Jeshlee VijayakumarFlorence Besse
Sep 27, 2019·Frontiers in Cellular Neuroscience·Francesca Mattedi, Alessio Vagnoni
Jan 14, 2021·The Journal of Comparative Neurology·Alexandre DumoulinEsther T Stoeckli

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