Related Experiment Video
Updated: Jul 18, 2025
Neurobehavioral Assessments in a Mouse Model of Neonatal Hypoxic-ischemic Brain Injury
Published on: November 24, 2017
The RNA m6A modification might participate in microglial activation during hypoxic-ischemic brain damage in neonatal
Xiaojuan Su1, Lingyi Huang1,2, Shiping Li1
1Department of Pediatrics/Key Laboratory of Birth Defects and Related Diseases of Women and Children (Ministry of Education), West China Second University Hospital, Sichuan University, Chengdu, 610041, China.
Background:
The RNA m6A modification has been implicated in multiple neurological diseases as well as macrophage activation. However, whether it regulates microglial activation during hypoxic-ischemic brain damage (HIBD) in neonates remains unknown. Here, we aim to examine whether the m6A modification is involved in modulating microglial activation during HIBD. We employed an oxygen and glucose deprivation microglial model for in vitro studies and a neonatal mouse model of HIBD. The brain tissue was subjected to RNA-seq to screen for significant changes in the mRNA m6A regulator. Thereafter, we performed validation and bioinformatics analysis of the major m6A regulators.
Results:
RNA-seq analysis revealed that, among 141 m6A regulators, 31 exhibited significant differential expression (FC (abs) ≥ 2) in HIBD mice. We then subjected the major m6A regulators Mettl3, Mettl14, Fto, Alkbh5, Ythdf1, and Ythdf2 to further validation, and the results showed that all were significantly downregulated in vitro and in vivo. GO analysis reveals that regulators are mainly involved in the regulation of cellular and metabolic processes. The KEGG results indicate the involvement of the signal transduction pathway.
Conclusions:
Our findings demonstrate that m6A modification of mRNA plays a crucial role in the regulation of microglial activation in HIBD, with m6A-associated regulators acting as key modulators of microglial activation.
Insights
This study investigates the role of N6-methyladenosine (m6A) modification in neonatal hypoxic-ischemic brain damage (HIBD). Key m6A regulators were found to be downregulated, highlighting their crucial role in microglial activation during HIBD.
Area of Science:
- Neuroscience
- Molecular Biology
- Developmental Biology
Background:
- N6-methyladenosine (m6A) RNA modification is linked to neurological diseases and macrophage activation.
- The role of m6A in microglial activation during neonatal hypoxic-ischemic brain damage (HIBD) is currently unknown.
Purpose of the Study:
- To investigate the involvement of m6A modification in regulating microglial activation during HIBD.
- To identify key m6A regulators modulated during HIBD.
Main Methods:
- Utilized an in vitro oxygen and glucose deprivation microglial model.
- Employed a neonatal mouse model of HIBD.
- Conducted RNA-sequencing (RNA-seq) on brain tissue to identify differentially expressed m6A regulators, followed by validation and bioinformatics analysis.
Main Results:
- RNA-seq identified 31 significantly differentially expressed m6A regulators out of 141.
- Key regulators including Mettl3, Mettl14, Fto, Alkbh5, Ythdf1, and Ythdf2 were significantly downregulated both in vitro and in vivo.
- Gene Ontology (GO) and KEGG pathway analyses indicated involvement in cellular, metabolic, and signal transduction processes.
Conclusions:
- mRNA m6A modification plays a critical role in regulating microglial activation in the context of HIBD.
- m6A-associated regulators are identified as key modulators of microglial activation during HIBD.
