Related Experiment Video
Updated: Aug 6, 2026
Animal Models of Depression - Chronic Despair Model (CDM)
Published on: September 23, 2021
Microglial Sestrin2 alleviates depressive-like behaviors and cognitive impairment in a YTHDF1-dependent manner
Ming Sun1, Jiaming Shen2, Rongrong Huang3
1Department of Ultrasound, Affiliated Hospital of Nantong University, Nantong, China.
Objective:
This study aims to investigate the role of microglial Sestrin2 in chronic unpredictable stress (CUS)-induced depressive-like behaviors and cognitive impairment in mice, and to explore the upstream molecular mechanism underlying the abnormal expression of microglial Sestrin2.
Methods:
Microglia-specific overexpression of Sestrin2 was achieved by injecting adeno-associated virus (AAV) into the CUS mouse hippocampus. Depressive-like behaviors were assessed using sucrose preference, tail suspension, and forced swim tests. Cognitive function was evaluated by the Morris water maze. Levels of IL-1β and IL-6 in the hippocampus and cell supernatants were measured by ELISA. BV2 microglial cells were used for in vitro mechanistic studies. YTHDF1 siRNA and overexpressive lentivirus were used to regulate YTHDF1 expression in vitro. RNA immunoprecipitation was performed to demonstrate the physical interaction between YTHDF1 and Sestrin2 mRNA.
Results:
Sestrin2 expression was significantly reduced in the hippocampus of CUS mice. Overexpression of Sestrin2 specifically in microglia ameliorated CUS-induced depressive-like behaviors, cognitive impairment, and inflammatory levels. YTHDF1 expression was also reduced in the CUS hippocampus. Mechanistically, YTHDF1 bound to Sestrin2 mRNA and knockdown of YTHDF1 decreased Sestrin2 expression. Molecular biology prediction results showed that positions 1943 and 2,114 of Sestrin2 mRNA are high-confidence N6-methyladenosine (m6A) modification sites. Mutation of the 2,114 site on Sestrin2 mRNA inhibited the effect of YTHDF1 on 3Flag expression. Furthermore, YTHDF1 knockdown promoted IL-1β and IL-6 production in BV2 cells, which was reversed by Sestrin2 overexpression.
Conclusion:
Microglial Sestrin2 alleviates depressive-like behaviors, cognitive impairment and neuroinflammation. YTHDF1 regulates Sestrin2 expression via an m6A-dependent mechanism, and the YTHDF1-Sestrin2 axis may represent a novel therapeutic target for major depressive disorder.
Insights
Microglial Sestrin2 overexpression alleviates depression-like behaviors and cognitive deficits in mice. The YTHDF1 protein regulates Sestrin2 expression through an m6A-dependent mechanism, offering a potential therapeutic target for major depressive disorder.
Area of Science:
- Neuroscience
- Molecular Biology
- Immunology
Background:
- Chronic unpredictable stress (CUS) induces depressive-like behaviors and cognitive impairment.
- Microglia play a crucial role in neuroinflammation associated with depression.
- The precise molecular mechanisms regulating microglial function in stress-induced disorders remain incompletely understood.
Purpose of the Study:
- To investigate the role of microglial Sestrin2 in CUS-induced depressive-like behaviors and cognitive impairment.
- To elucidate the upstream molecular mechanism controlling microglial Sestrin2 expression.
Main Methods:
- Mice were subjected to CUS, and microglia-specific Sestrin2 overexpression was induced using adeno-associated virus (AAV) vectors.
- Behavioral tests (sucrose preference, tail suspension, forced swim, Morris water maze) assessed depressive-like behaviors and cognitive function.
- ELISA measured inflammatory cytokines (IL-1β, IL-6).
- In vitro studies in BV2 microglial cells utilized YTHDF1 siRNA and overexpression, alongside RNA immunoprecipitation (RIP) and m6A site mutation analysis.
Main Results:
- CUS significantly reduced Sestrin2 expression in the mouse hippocampus.
- Microglial Sestrin2 overexpression ameliorated CUS-induced depressive-like behaviors, cognitive deficits, and neuroinflammation.
- YTHDF1 expression was also reduced by CUS, and YTHDF1 directly bound to Sestrin2 mRNA in an m6A-dependent manner.
- YTHDF1 knockdown increased pro-inflammatory cytokine production, an effect reversed by Sestrin2 overexpression.
Conclusions:
- Microglial Sestrin2 exerts protective effects against CUS-induced depressive-like behaviors, cognitive impairment, and neuroinflammation.
- The YTHDF1 protein regulates Sestrin2 expression via an m6A-dependent mechanism.
- The YTHDF1-Sestrin2 axis represents a promising therapeutic target for major depressive disorder.
