Immunoprecipitation of Ribosome-Bound mRNAs from Astrocytic Perisynaptic Processes of the Mouse Hippocampus

Noémie Mazaré; Marc Oudart; Giselle Cheung; Anne-Cécile Boulay; Martine Cohen-Salmon

doi:10.1016/j.xpro.2020.100198

Immunoprecipitation of Ribosome-Bound mRNAs from Astrocytic Perisynaptic Processes of the Mouse Hippocampus

STAR Protoc. 2020 Dec 4;1(3):100198. doi: 10.1016/j.xpro.2020.100198. eCollection 2020 Dec 18.

Authors

Noémie Mazaré¹, Marc Oudart¹, Giselle Cheung², Anne-Cécile Boulay¹, Martine Cohen-Salmon¹

Affiliations

¹ Physiology and Physiopathology of the Gliovascular Unit Research Group, Center for Interdisciplinary Research in Biology (CIRB), College de France, CNRS Unité Mixte de Recherche 724, INSERM Unité 1050, Labex Memolife, PSL Research University, Paris, France.
² Neuroglial Interactions in Cerebral Physiopathology Research Group, Center for Interdisciplinary Research in Biology (CIRB), College de France, CNRS Unité Mixte de Recherche 724, INSERM Unité 1050, Labex Memolife, PSL Research University, Paris, France.

Abstract

Translation of distally localized mRNAs is an evolutionary mechanism occurring in polarized cells. It has been observed in astrocytes, whose processes contact blood vessels and synapses. Here, we describe a protocol for the purification of the entire pool of ribosome-bound mRNAs in perisynaptic astrocytic processes (PAPs). Our procedure combines the preparation of synaptogliosomes with a refined translating ribosome affinity purification technique. This approach can be used in any brain region to probe the physiological relevance of local translation in PAPs. For complete details on the use and execution of this protocol, please refer to Mazaré et al. (2020).

Keywords: Molecular Biology; Neuroscience.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Astrocytes / pathology
Astrocytes / physiology
Biophysical Phenomena
Cell Communication
Hippocampus / physiology
Immunoprecipitation / methods*
Mice
Phagocytosis
RNA, Messenger / isolation & purification*
Ribosomes / genetics
Ribosomes / metabolism
Synapses / physiology

Substances

RNA, Messenger