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Astrocytes pp 105-116 | Cite as

Preparing the Astrocyte Perivascular Endfeet Transcriptome to Investigate Astrocyte Molecular Regulations at the Brain–Vascular Interface

  • Anne-Cécile Boulay
  • Noémie Mazaré
  • Bruno Saubaméa
  • Martine Cohen-SalmonEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1938)

Abstract

Astrocytes send out long processes that are terminated by endfeet at the vascular surface and regulate vascular functions in particular through the expression of a specific molecular repertoire in perivascular endfeet. We recently proposed that local translation might sustain this structural and functional polarization. More specifically we showed that a subset of mRNAs is distributed in astrocyte endfeet and characterized this transcriptome. We also identified among these endfeet RNAs, the ones bound to ribosomes, the polysomal astrocyte endfeet mRNAs, which we called the endfeetome. Here, we describe experimental strategies to identify mRNAs and polysomes in astrocyte perivascular endfeet, which are based on the combination of gliovascular unit purification and astrocyte-specific translating ribosome affinity purification.

Key words

Astrocyte Endfeet Brain vessels Gliovascular unit Transcriptome Translating ribosome affinity purification (TRAP) 

Notes

Acknowledgments

This work has received support under the program “Investissements d’Avenir” launched by the French Government and implemented by the ANR, with the references: ANR-10-LABX-54 MEMO LIFE, ANR-11-IDEX-0001-02 PSL* Research University and by Fondation pour la recherche médicale (AJE20171039094).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Anne-Cécile Boulay
    • 1
    • 2
  • Noémie Mazaré
    • 1
    • 2
  • Bruno Saubaméa
    • 3
    • 4
    • 5
  • Martine Cohen-Salmon
    • 1
    • 2
    Email author
  1. 1.Collège de France, Center for Interdisciplinary Research in Biology (CIRB)/Centre National de la Recherche Scientifique CNRS, Unité Mixte de Recherche 7241/Institut National de la Santé et de la Recherche Médicale INSERM, U1050/75231Paris Cedex 05France
  2. 2.Paris Science Lettre Research UniversityParisFrance
  3. 3.Institut National de la Santé et de la Recherche Médicale INSERM U1144ParisFrance
  4. 4.Université Paris Descartes, UMR-S 1144ParisFrance
  5. 5.Université Paris Diderot, UMR-S 1144ParisFrance

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