Preparing the Astrocyte Perivascular Endfeet Transcriptome to Investigate Astrocyte Molecular Regulations at the Brain–Vascular Interface
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 wordsAstrocyte Endfeet Brain vessels Gliovascular unit Transcriptome Translating ribosome affinity purification (TRAP)
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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