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Updated: Aug 3, 2025
Isolation of Region-specific Microglia from One Adult Mouse Brain Hemisphere for Deep Single-cell RNA Sequencing
Published on: December 3, 2019
Integrative Analysis of Single-Cell and Bulk Sequencing Data Depicting the Expression and Function of P2ry12 in
Chenglong Wang1,2,3, Li Peng1,2,3, Yuan Wang1,2,3
1Department of Pathology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, China.
Abstract:
P2ry12 is a microglial marker gene. Recently, increasing evidence has demonstrated that its expression levels can vary in response to different CNS disorders and can affect microglial functions, such as polarization, plasticity, and migration. However, the expression and function of P2ry12 in microglia during ischemia-reperfusion injury (IRI) remain unclear. Here, we developed a computational method to obtain microglia-specific P2ry12 genes (MSPGs) using sequencing data associated with IRI. We evaluated the change in comprehensive expression levels of MSPGs during IRI and compared it to the expression of P2ry12 to determine similarity. Subsequently, the MSPGs were used to explore the P2ry12 functions in microglia through bioinformatics. Moreover, several animal experiments were also conducted to confirm the reliability of the results. The expression of P2ry12 was observed to decrease gradually within 24 h post injury. In response, microglia with reduced P2ry12 expression showed an increase in the expression of one receptor-encoding gene (Flt1) and three ligand-encoding genes (Nampt, Igf1, and Cxcl2). Furthermore, double-labeling immunofluorescence staining revealed that inhibition of P2ry12 blocked microglial migration towards vessels during IRI. Overall, we employ a combined computational and experimental approach to successfully explore P2ry12 expression and function in microglia during IRI.
Insights
P2ry12 expression in microglia decreases during ischemia-reperfusion injury (IRI). Reduced P2ry12 impacts microglial function and migration, suggesting a role in CNS disorders.
Area of Science:
- Neuroscience
- Immunology
- Computational Biology
Background:
- P2ry12 is a key microglial marker gene.
- Its expression is altered in various central nervous system (CNS) disorders.
- P2ry12 influences microglial functions like polarization, plasticity, and migration.
Purpose of the Study:
- To investigate the expression and function of P2ry12 in microglia during ischemia-reperfusion injury (IRI).
- To identify microglia-specific P2ry12 genes (MSPGs) associated with IRI.
- To explore the functional role of P2ry12 in microglial responses to IRI.
Main Methods:
- Development of a computational method to identify MSPGs from IRI sequencing data.
- Evaluation of MSPG expression changes during IRI.
- Bioinformatic analysis to explore P2ry12 functions.
- Animal experiments including double-labeling immunofluorescence staining.
Main Results:
- P2ry12 expression gradually decreased within 24 hours post-IRI.
- Reduced P2ry12 correlated with increased expression of Flt1, Nampt, Igf1, and Cxcl2 in microglia.
- Inhibition of P2ry12 impaired microglial migration towards vessels during IRI.
Conclusions:
- Combined computational and experimental approaches elucidated P2ry12 expression and function in microglia during IRI.
- P2ry12 plays a role in regulating microglial migration and gene expression post-IRI.
- Findings provide insights into microglial behavior in CNS injury.
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