Abstract
Axonal transport is used by neurons to distribute mRNAs, proteins, and organelles to their peripheral compartments in order to sustain their structural and functional integrity. Cargoes are transported along the microtubule (MT) network whose post-translational modifications influence transport dynamics. Here, we describe methods to modulate MT acetylation and record its impact on axonal transport in cultured mouse cortical projection neurons as well as in motoneurons of Drosophila melanogaster third-instar larvae. Specifically, we provide a step-by step procedure to reduce the level of MT acetylation and to record and analyze the transport of dye-labeled organelles in projection neuron axons cultured in microfluidic chambers. In addition, we describe the method to record and analyze GFP-tagged mitochondria transport along the motoneuron axons of transgenic Drosophila melanogaster third-instar larvae.
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Acknowledgments
S.T. and R.L.B are PhD students from F.R.S-F.N.R.S.; L.B. and L.N. are respectively Postdoctoral Researcher and Director from F.R.S-F.N.R.S. The work in the Nguyen laboratory is supported by the F.R.S.-F.N.R.S. (Synet; EOS 0019118F-RG36), the Fonds Leon Fredericq, the Fondation Médicale Reine Elisabeth, the Fondation Simone et Pierre Clerdent, the Belgian Science Policy (IAP-VII network P7/20), and the ERANET Neuron STEM-MCD and NeuroTalk. The figures were made with Biorender (https://biorender.com/).
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Turchetto, S., Le Bail, R., Broix, L., Nguyen, L. (2022). Molecular Analysis of Axonal Transport Dynamics upon Modulation of Microtubule Acetylation. In: Vagnoni, A. (eds) Axonal Transport. Methods in Molecular Biology, vol 2431. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1990-2_10
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DOI: https://doi.org/10.1007/978-1-0716-1990-2_10
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