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Microtubules pp 375–383Cite as

Cryo-EM Visualization of Neuronal Particles Inside Microtubules

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2430))

Abstract

Neuronal microtubules have long been known to contain intraluminal particles, called MIPs (microtubule inner proteins), most likely involved in the extreme stability of microtubules in neurons. This chapter describes a cryo-electron microscopy-based assay to visualize microtubules containing neuronal MIPs. We present two protocols to prepare MIPs-containing microtubules, using either in vitro microtubule polymerization assays or extraction of microtubules from mouse hippocampal neurons in culture.

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Acknowledgments

We thank Lisa De Macedo and Nicolas Chaumontel for technical help in protein purification and culture of neurons. We thank Naïs Martin for help in Fig. 1 scheme drawing. This work was supported by INSERM, CNRS and Grenoble Alpes University, and by grants from the French Research National Agency (2017-CE11-0026 MAMAs). This work used the EM facilities at the Grenoble Instruct-ERIC Center (ISBG; UMS 3518 CNRS CEA-UGA-EMBL) with support from the French Infrastructure for Integrated Structural Biology (FRISBI; ANR-10-INSB-05-02) and GRAL, a project of the University Grenoble Alpes graduate school (Ecoles Universitaires de Recherche) CBH-EUR-GS (ANR-17-EURE-0003) within the Grenoble Partnership for Structural Biology. The IBS Electron Microscope facility is supported by the Auvergne Rhône-Alpes Region, the Fonds Feder, the Fondation pour la Recherche Médicale and GIS-IBiSA. We thank the IBS facility members for their help.

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Author notes

  1. Sylvie Gory-Fauré and Julie Delaroche contributed equally to this work.

Authors and Affiliations

  1. Univ. Grenoble Alpes, Inserm U1216, CNRS, Grenoble Institut Neurosciences, Grenoble, France

    Sylvie Gory-Fauré, Julie Delaroche, Camille Cuveillier, Christian Delphin & Isabelle Arnal

Authors
  1. Sylvie Gory-Fauré
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  2. Julie Delaroche
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  3. Camille Cuveillier
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  4. Christian Delphin
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  5. Isabelle Arnal
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Corresponding author

Correspondence to Isabelle Arnal .

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Editors and Affiliations

  1. Dept Chemistry and Biotechnology, Tottori University, Tottori, Japan

    Hiroshi Inaba

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Gory-Fauré, S., Delaroche, J., Cuveillier, C., Delphin, C., Arnal, I. (2022). Cryo-EM Visualization of Neuronal Particles Inside Microtubules. In: Inaba, H. (eds) Microtubules. Methods in Molecular Biology, vol 2430. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1983-4_24

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  • DOI: https://doi.org/10.1007/978-1-0716-1983-4_24

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1982-7

  • Online ISBN: 978-1-0716-1983-4

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