Related Experiment Video
Updated: Jun 17, 2026
Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice
Published on: December 29, 2016
miR-497 regulates neuronal death in mouse brain after transient focal cerebral ischemia
Ke-Jie Yin1, Zhen Deng, Huarong Huang
1Cardiovascular Center, Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor, MI 48109, USA.
Abstract:
Dysfunction of the microRNA (miR) network has been emerging as a major regulator in neurological diseases. However, little is known about the functional significance of unique miRs in ischemic brain damage. Here, we found that miR-497 is induced in mouse brain after transient middle cerebral artery occlusion (MCAO) and mouse N2A neuroblastoma (N2A) cells after oxygen-glucose deprivation (OGD). Loss-of-miR-497 function significantly suppresses OGD-induced N2A cell death, whereas gain-of-miR-497 function aggravates OGD-induced neuronal loss. Moreover, miR-497 directly binds to the predicted 3'-UTR target sites of bcl-2/-w genes. Furthermore, knockdown of cerebral miR-497 effectively enhances bcl-2/-w protein levels in the ischemic region, attenuates ischemic brain infarction, and improves neurological outcomes in mice after focal cerebral ischemia. Taken together, our data suggest that miR-497 promotes ischemic neuronal death by negatively regulating antiapoptotic proteins, bcl-2 and bcl-w. We raise the possibility that this pathway may contribute to the pathogenesis of the ischemic brain injury in stroke.
Insights
MicroRNA-497 (miR-497) exacerbates ischemic brain damage by targeting bcl-2 and bcl-w. Reducing miR-497 levels protects against stroke-induced neuronal death and improves outcomes in mice.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- MicroRNA (miR) network dysfunction is implicated in neurological diseases.
- The specific role of individual miRs in ischemic brain injury remains largely unknown.
Purpose of the Study:
- To investigate the functional significance of miR-497 in ischemic brain damage.
- To elucidate the molecular mechanisms by which miR-497 influences neuronal survival after ischemia.
Main Methods:
- Induction of miR-497 in mouse models of focal cerebral ischemia (transient middle cerebral artery occlusion) and in vitro (oxygen-glucose deprivation in N2A cells).
- Manipulation of miR-497 levels (loss-of-function and gain-of-function) to assess its impact on neuronal survival.
- Identification of miR-497 targets using bioinformatics and experimental validation (luciferase reporter assays, Western blotting).
- Evaluation of neurological outcomes and infarct volume in mice following miR-497 knockdown.
Main Results:
- miR-497 expression is upregulated in the mouse brain and neuronal cells following ischemic conditions.
- Loss of miR-497 function protects against oxygen-glucose deprivation-induced cell death, while its overexpression exacerbates neuronal loss.
- miR-497 directly targets the 3'-untranslated regions of bcl-2 and bcl-w genes.
- Knockdown of miR-497 in the ischemic brain increases bcl-2/-w protein levels, reduces infarct size, and improves neurological function.
Conclusions:
- miR-497 promotes ischemic neuronal death by negatively regulating the antiapoptotic proteins bcl-2 and bcl-w.
- The miR-497/bcl-2/-w pathway is a potential contributor to the pathogenesis of ischemic brain injury in stroke.
- Targeting miR-497 may represent a therapeutic strategy for stroke treatment.
