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Isolation of Region-specific Microglia from One Adult Mouse Brain Hemisphere for Deep Single-cell RNA Sequencing
Published on: December 3, 2019
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Fixed single-cell transcriptomic characterization of human radial glial diversity.
Elliot R Thomsen1, John K Mich1, Zizhen Yao1
1Allen Institute for Brain Science, Seattle, Washington, USA.
Nature Methods
|November 3, 2015
Summary
Researchers developed a new method, FRISCR, to profile rare human neocortical progenitors. This technique identified distinct markers for radial glia (RG) subtypes, advancing our understanding of brain development.
Area of Science:
- Neuroscience
- Developmental Biology
- Genomics
Background:
- Characterizing diverse progenitors in human neocortical development is challenging due to their rarity and reliance on combined markers.
- Radial glia (RG) are crucial progenitors but difficult to isolate and study.
- Existing methods struggle with rare cell populations and require live cells.
Purpose of the Study:
- To develop a novel method for profiling rare, fixed, and stained individual cells.
- To identify molecular markers and profiles for distinct human neocortical radial glia (RG) subtypes.
- To advance the understanding of human neocortical progenitor development.
Main Methods:
- Developed Fixed and Recovered Intact Single-cell RNA (FRISCR) technology.
- Applied FRISCR to profile transcriptomes of individual, fixed, stained, and sorted primary human RG.
- Utilized cell sorting based on morphology and staining for progenitor isolation.
Main Results:
- FRISCR successfully profiled rare RG cells (1% of midgestation cortex).
- Identified ventricular zone-enriched RG (vRG) expressing ANXA1 and CRYAB.
- Identified outer subventricular zone-localized RG (oRG) expressing HOPX.
Conclusions:
- FRISCR enables targeted single-cell transcriptomic profiling of any fixed tissue, even without live-cell markers.
- The study identified specific molecular markers (ANXA1, CRYAB, HOPX) for vRG and oRG subtypes.
- These findings are essential for understanding human neocortical progenitor diversity and development.
