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Updated: May 23, 2026
Evaluating Cell Death Signaling by Immunofluorescence in a Rat Model of Ischemic Stroke
Published on: January 3, 2025
Dynamic changes of mitochondrial fission proteins after transient cerebral ischemia in mice
Wentao Liu1, Fengfeng Tian, Tomoko Kurata
1Department of Neurology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 2-5-1 Shikatacho, Okayama 700-8558, Japan.
Abstract:
With fusion or fission, mitochondria alter their morphology in response to various physiological and pathological stimuli resulting in either elongated, tubular, interconnected or fragmented form. Immunohistochemistry and Western blot analyses were performed at 2, 7, 14 and 28 d after 90 min of transient middle cerebral artery occlusion (tMCAO) in mice. The present study showed that mitochondrial fission protein fission 1 (Fis1) and phosphorylated dynamin-related protein 1 (P-Drp1) both progressively increased with the peak at 14 d after tMCAO. Double immunofluorescent analysis showed the number of double positive cells with Fis1/Drp1 reduced between 2 and 28 d after 90 min of tMCAO, and also showed some double positive cells with Fis1/terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) in the peri-infract regions at 2d after the reperfusion. The present study suggests a progressive activation of mitochondrial fission proteins Fis1 and P-Drp1 in relation to apoptotic process in neural cells of the peri-infract regions after tMCAO.
Insights
Mitochondrial fission proteins Fis1 and P-Drp1 increase after stroke, peaking at 14 days. This suggests their activation is linked to neural cell apoptosis in stroke-affected brain regions.
Area of Science:
- Neuroscience
- Cell Biology
- Pathology
Background:
- Mitochondria dynamically change shape through fusion and fission.
- These morphological changes are crucial for cellular function and survival.
- Mitochondrial dynamics are implicated in various physiological and pathological conditions.
Purpose of the Study:
- To investigate the role of mitochondrial fission proteins in neural cells following transient middle cerebral artery occlusion (tMCAO).
- To determine the temporal changes in mitochondrial fission protein expression and localization after ischemic stroke.
- To explore the relationship between mitochondrial fission and apoptosis in the peri-infarct region.
Main Methods:
- Mice underwent 90 minutes of transient middle cerebral artery occlusion (tMCAO).
- Immunohistochemistry and Western blot analyses were performed at 2, 7, 14, and 28 days post-tMCAO.
- Double immunofluorescence staining was used to detect Fis1, Drp1, and TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling).
Main Results:
- Mitochondrial fission proteins, fission 1 (Fis1) and phosphorylated dynamin-related protein 1 (P-Drp1), progressively increased after tMCAO, peaking at 14 days.
- The co-localization of Fis1 and Drp1 decreased between 2 and 28 days post-tMCAO.
- Fis1 and TUNEL-positive cells were observed in the peri-infarct regions at 2 days post-reperfusion, indicating apoptosis.
Conclusions:
- A progressive activation of mitochondrial fission proteins Fis1 and P-Drp1 occurs in neural cells following ischemic stroke.
- Mitochondrial fission is associated with the apoptotic process in the peri-infarct regions after tMCAO.
- These findings highlight the involvement of mitochondrial dynamics in stroke-induced neural cell death.
