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Updated: Oct 29, 2025
Isolation of Cortical Microglia with Preserved Immunophenotype and Functionality From Murine Neonates
Published on: January 30, 2014
The potential roles of m6A modification in regulating the inflammatory response in microglia
Qi Li1, Shaohong Wen1, Weizhen Ye1
1China National Clinical Research Center for Neurological Diseases, Beijing Tiantan Hospital, Capital Medical University, 119 South Fourth Ring West Road, Fengtai District, Beijing, 100070, People's Republic of China.
Background:
Microglia are key regulators of the inflammatory response in the brain. Adenosine in RNAs can be converted to m6A (N6-methyladenosine), which regulates RNA metabolism and functions as a key epitranscriptomic modification. The m6A modification pattern and m6A-related signatures under pro-inflammatory and anti-inflammatory conditions of microglia remain unclear.
Methods:
Primary rat microglia were differentiated into pro-inflammatory M1-like (M1-L), anti-inflammatory M2-like (M2-L), and resting, unstimulated (M0-L) phenotypes. m6A mRNA and lncRNA epitranscriptomic microarray analyses were performed, and pathway analysis was conducted to understand the functional implications of m6A methylation in mRNAs and lncRNAs. The m6A methylation level and gene expression of mRNAs and lncRNAs were subsequently verified by m6A Me-RIP and qRT-PCR.
Results:
A total of 1588 mRNAs and 340 lncRNAs, 315 mRNAs and 38 lncRNAs, and 521 mRNAs and 244 lncRNAs were differentially m6A methylated between M1-L and M0-L (M1-L/M0-L), M2-L and M0-L (M2-L/M0-L), M2-L and M1-L (M2-L/M1-L), respectively. Furthermore, 4902 mRNAs, 4676 mRNAs, and 5095 mRNAs were identified distinctively expressed in M1-L/M0-L, M2-L/M0-L, and M2-L/M1-L, respectively. Pathway analysis of differentially m6A methylated mRNAs and lncRNAs in M1-L/M0-L identified immune system, signal transduction, and protein degradation processes. In contrast, the distinct m6A methylated mRNAs in M2-L/M0-L were involved in genetic information processing, metabolism, cellular processes, and neurodegenerative disease-related pathways. We validated m6A methylation and the expression levels of five mRNAs and five lncRNAs, which were involved in upregulated pathways in M1-L/M0-L, and five mRNAs involved in upregulated pathways in M2-L/M0-L.
Conclusions:
These findings identify a distinct m6A epitranscriptome in microglia, and which may serve as novel and useful regulator during pro-inflammatory and anti-inflammatory response of microglia.
Insights
Microglia
Area of Science:
- Neuroscience
- Immunology
- Molecular Biology
Background:
- Microglia are central to brain inflammation.
- N6-methyladenosine (m6A) is a key RNA modification impacting RNA metabolism.
- The role of m6A in microglial inflammatory states is not well understood.
Purpose of the Study:
- To investigate the m6A epitranscriptome in different microglial phenotypes.
- To identify m6A-related signatures in pro-inflammatory and anti-inflammatory microglia.
- To understand the functional implications of m6A modifications in microglia.
Main Methods:
- Primary rat microglia were differentiated into M1-like (pro-inflammatory), M2-like (anti-inflammatory), and M0-like (resting) states.
- Epitranscriptomic microarray analysis was used to profile m6A modification patterns in mRNAs and lncRNAs.
- m6A Me-RIP and qRT-PCR were employed for validation of key findings.
Main Results:
- Significant differences in m6A methylation and expression were observed across microglial phenotypes.
- Pathway analysis revealed distinct functions for m6A-modified transcripts in M1-like and M2-like microglia.
- Immune and signaling pathways were prominent in pro-inflammatory microglia, while metabolic and neurodegenerative pathways were enriched in anti-inflammatory microglia.
Conclusions:
- A distinct m6A epitranscriptome exists in microglia.
- m6A modification patterns are unique to different microglial activation states.
- The m6A epitranscriptome represents a potential regulatory mechanism in microglial inflammatory responses.
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