Purification of astrocyte subtype based on a double reporter mice approach for downstream transcription profiling

Marion Aguirrebengoa; David Ohayon

doi:10.1016/j.xpro.2021.101009

Purification of astrocyte subtype based on a double reporter mice approach for downstream transcription profiling

STAR Protoc. 2021 Dec 10;2(4):101009. doi: 10.1016/j.xpro.2021.101009. eCollection 2021 Dec 17.

Authors

Marion Aguirrebengoa¹, David Ohayon²

Affiliations

¹ BigA Core Facility, Centre de Biologie Intégrative, Université de Toulouse, 118 Route de Narbonne, 31062 Toulouse, France.
² Molecular, Cellular and Developmental Biology Department (MCD) UMR 5077 CNRS, Centre de Biologie Intégrative (CBI), Université Paul Sabatier, 118 Route de Narbonne, 31062 Toulouse, France.

Abstract

Characterizing the molecular signature of a cell subtype leads to a better understanding of cell diversity, as this molecular data can identify new cellular markers and offer insights about cell function. Here, we describe an efficient protocol to separate a subtype of astrocytes, the Olig2-AS, from other glial cells by using a double reporter mouse approach and to determine the transcriptome profile of the Olig2-AS from the postnatal spinal cord using RNA-sequencing analysis. For complete details on the use and execution of this protocol, please refer to Ohayon et al. (2021).

Keywords: Cell Biology; Cell isolation; Flow Cytometry/Mass Cytometry; Gene Expression; Model Organisms; Neuroscience; RNAseq; Sequence analysis.

Publication types

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

MeSH terms

Animals
Astrocytes / chemistry
Astrocytes / cytology*
Astrocytes / metabolism
Flow Cytometry / methods*
Mice
Mice, Transgenic
Neuroglia / cytology
RNA-Seq / methods*
Spinal Cord / cytology
Transcriptome / genetics*