Molecular Analysis of Axonal Transport Dynamics upon Modulation of Microtubule Acetylation.

ATAT1; Axonal transport; Cargo; HDAC6; MT acetylation; Organelles; Acetylation; Mice; Motor Neurons; Axonal Transport/physiology; Axons; Drosophila melanogaster; Microtubules

Abstract :

[en] 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.

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

Turchetto, Silvia ^✱; Université de Liège - ULiège > Département des sciences biomédicales et précliniques

Le Bail, Romain ^✱; Université de Liège - ULiège > GIGA

Broix, Loïc ; Université de Liège - ULiège > GIGA > GIGA Stem Cells - Molecular Regulation of Neurogenesis

Nguyen, Laurent ; Université de Liège - ULiège > GIGA > GIGA Stem Cells

^✱ These authors have contributed equally to this work.

Language :

English

Title :

Molecular Analysis of Axonal Transport Dynamics upon Modulation of Microtubule Acetylation.

Publication date :

2022

Journal title :

Methods in Molecular Biology

ISSN :

1064-3745

eISSN :

1940-6029

Publisher :

Humana Press Inc., United States

Volume :

2431

Pages :

207-224

Peer reviewed :

Peer Reviewed verified by ORBi

Funding text :

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 M?dicale 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/).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 Fon-dation 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/).

Data Set :

10.1007/978-1-0716-1990-2_10

Available on ORBi :

since 24 May 2022

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