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Induction of phosphorylated-Stat3 following focal cerebral ischemia in mice
1Department of Anatomy, Ehime University School of Medicine, Shigenobu, 791-0295, Ehime, Japan.
Abstract:
It has been shown that Stat3 is induced following transient cerebral ischemia in rat. However there is no evidence that cerebral ischemia stimulates the expression of phosphorylated-Stat3 (p-Stat3), which can activate cytokine-mediated signal transduction from the membrane to the nucleus. In the present study, we investigated the changes in p-Stat3 expression following middle cerebral artery occlusion in mice. Western blot analysis revealed a significant increase in the p-Stat3 protein in the peripheral part of the ischemic area, starting from 6 h after ischemia. p-Stat3 immunoreactivity was detected only in neurons, but not in astrocytes or microglia, and p-Stat3-positive neurons were increased in number in the peripheral part of the ischemic area at 24 h after ischemia. Double staining with aTdT-mediated biotinylated UTP nick end labeling (TUNEL) kit and the p-Stat3 antibody indicated that p-Stat3-positive neurons were also TUNEL-positive. Subsequent immuno-electron microscopic observations showed that p-Stat3-positive neurons were at different stages of degeneration. The present findings suggest that the increased expression of p-Stat3 after cerebral ischemia could play a crucial role in ischemia-induced neuron death.
Insights
Phosphorylated-Stat3 (p-Stat3) expression increases in neurons after cerebral ischemia, indicating its role in neuron death. This finding highlights p-Stat3 as a potential target for stroke therapies.
Area of Science:
- Neuroscience
- Molecular Biology
- Pathology
Background:
- Signal transducer and activator of transcription 3 (Stat3) is induced by transient cerebral ischemia.
- The role of phosphorylated-Stat3 (p-Stat3) in cerebral ischemia-induced neuronal damage remains unclear.
- p-Stat3 activation is crucial for cytokine-mediated signal transduction.
Purpose of the Study:
- To investigate the expression and localization of p-Stat3 following middle cerebral artery occlusion (MCAO) in mice.
- To determine the association between p-Stat3 expression and neuronal cell death after ischemic stroke.
Main Methods:
- Middle cerebral artery occlusion (MCAO) model in mice.
- Western blot analysis to detect p-Stat3 protein levels.
- Immunohistochemistry and immunoelectron microscopy to visualize p-Stat3 localization.
- TUNEL assay to identify apoptotic neurons.
Main Results:
- p-Stat3 protein significantly increased in the ischemic area starting 6 hours post-MCAO.
- p-Stat3 immunoreactivity was exclusively observed in neurons, not glial cells.
- The number of p-Stat3-positive neurons increased by 24 hours post-MCAO.
- p-Stat3-positive neurons exhibited features of degeneration and were TUNEL-positive, indicating apoptosis.
Conclusions:
- Increased p-Stat3 expression in neurons following cerebral ischemia is associated with neuronal cell death.
- p-Stat3 may play a critical role in the pathogenesis of ischemia-induced neuronal death.
- Targeting p-Stat3 signaling could be a potential therapeutic strategy for stroke.
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