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Measuring the Impact of Tubulin Posttranslational Modifications on Axonal Transport

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Cytoskeleton Dynamics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2101))

Abstract

Axonal transport is a process essential for neuronal function and survival that takes place on the cellular highways—the microtubules. It requires three major components: the microtubules that serve as tracks for the transport, the motor proteins that drive the movement, and the transported cargoes with their adaptor proteins. Axonal transport could be controlled by tubulin posttranslational modifications, which by decorating specific microtubule tracks could determine the specificity of cargo delivery inside neurons. However, it appears that the effects of tubulin modifications on transport can be rather subtle, and might thus be easily overlooked depending on which parameter of the transport process is analyzed. Here we propose an analysis paradigm that allows detecting rather subtle alterations in neuronal transport, as induced for instance by accumulation of posttranslational polyglutamylation. Analyzing mitochondria movements in axons, we found that neither the average speed nor the distance traveled were affected by hyperglutamylation, but we detected an about 50% reduction of the overall motility, suggesting that polyglutamylation controls the efficiency of mitochondria transport in axons. Our protocol can readily be expanded to the analysis of the impact of other tubulin modifications on the transport of a range of different neuronal cargoes.

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Acknowledgments

This work was supported by the ANR-10-IDEX-0001-02, the LabEx CelTisPhyBio ANR-11-LBX-0038. CJ is supported by the Institut Curie, the French National Research Agency (ANR) awards ANR-12-BSV2-0007 and ANR-17-CE13-0021, the Institut National du Cancer (INCA) grant 2014-PL BIO-11-ICR-1, and the Fondation pour la Recherche Medicale (FRM) grant DEQ20170336756. MMM is supported by the EMBO short-term fellowship ASTF 148-2015 and by the Fondation Vaincre Alzheimer grant FR-16055p, and SB by the FRM grant FDT201805005465. We thank C. Alberti, E. Belloir, F. Bertrand, V. Dangles-Marie, I. Grandjean, C. Caspersen, H. Hermange, A. Thadal, G. Buhagiar, C. Serieyssol, S. Gadadhar, and M. Sittewelle (Institut Curie) for technical assistance. We are grateful to M.-N. Soler, C. Lovo, and L. Besse from the PICT-IBiSA@Orsay Imaging Facility of the Institut Curie supported by the ANR through the “Investment for the future” program (France-BioImaging, ANR-10-INSB-04), and to N. Manel (Institut Curie, Paris) for material and advice for the lentivirus production. We would like to thank F. Cordelières (Bordeaux Imaging Center, France) for the KymoToolBox plug-in, as well as M. Brill (Technical University Munich, Germany), F. Del Bene, V. Marthiens (Institut Curie), and C. González-Billault (University of Chile, Santiago, Chile) for instructive discussions and advice.

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Authors and Affiliations

  1. Institut Curie, PSL Research University, CNRS UMR3348, Centre Universitaire, Orsay, France

    Satish Bodakuntla, Maria M. Magiera & Carsten Janke

  2. Université Paris Sud, Université Paris-Saclay, CNRS UMR3348, Orsay, France

    Satish Bodakuntla, Maria M. Magiera & Carsten Janke

Authors
  1. Satish Bodakuntla
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  2. Maria M. Magiera
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  3. Carsten Janke
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Correspondence to Maria M. Magiera or Carsten Janke .

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  1. i3S University of Porto, Porto, Portugal

    Helder Maiato

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Bodakuntla, S., Magiera, M.M., Janke, C. (2020). Measuring the Impact of Tubulin Posttranslational Modifications on Axonal Transport. In: Maiato, H. (eds) Cytoskeleton Dynamics. Methods in Molecular Biology, vol 2101. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0219-5_20

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  • DOI: https://doi.org/10.1007/978-1-0716-0219-5_20

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0218-8

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