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Updated: Mar 24, 2026
Stereological and Flow Cytometry Characterization of Leukocyte Subpopulations in Models of Transient or Permanent Cerebral Ischemia
Published on: December 28, 2014
[Increased circulating Ly6Chigh monocyte subsets are correlated with cerebral infarct size in cerebral
Xin Zhang1, Hongxia Li2, Yuxiu Li2
1School of Graduates, Tianjin Medical University, Tianjin 300070; Tianjin Key Laboratory of Cardiovascular Remodeling and Target Organ Injury, Cardiovascular Disease Institute, Heart Center, Affiliated Hospital, Logistics University of the Chinese People's Armed Police Force, Tianjin 300162, China.
Objective:
To investigate the dynamic changes of monocyte subsets after cerebral ischemia/reperfusion in mice and their correlations with infarct size and neurological defects.
Methods:
Thirty male C57BL/6 mice were randomly divided into two groups: middle cerebral artery occlusion/reperfusion (MCAO/R) group and Sham group. MCAO/R mouse models were induced via the intraluminal suture technique (so called filament or suture model). The modified Neurological Severity Scores (mNSS) was used to assess neurological function 6, 12 hours, 1, 2, 3 days after MCAO/R. Blood samples were collected 1, 2 and 3 days after surgery to determine the dynamic changes of Ly6C(high) and Ly6C(low) subsets by flow cytometry. Triphenyltetrazolium chloride (TTC) staining and HE staining of the brains were also performed on day 1, 2 and 3. The relationships between the changes of monocyte subsets and the cerebral infarct size and neurological scores were studied by correlation analysis.
Results:
Compared with the baseline, the proportion of Ly6C(high) monocytes significantly increased 1 day after MCAO/R surgery, reached the peak level on the following day and then declined. Compared with the Sham group, the proportion of Ly6C(high) monocytes went up obviously at each time point. TTC staining revealed that the infarct size increased markedly 2 days after surgery. The proportion of Ly6C(high) monocytes in the MCAO/R group was positively associated with the percentage of cerebral infarct size, and it also showed a positive correlation with neurological function deficit scores.
Conclusion:
The dynamic changes of monocyte subsets after MCAO/R modeling revealed that Ly6C(high) subset peaked on day 2 after the operation and was correlated with cerebral infarct size.
Insights
Ly6C(high) monocytes increase after cerebral ischemia in mice, peaking on day 2. This monocyte subset correlates with larger infarct size and worse neurological deficits following middle cerebral artery occlusion/reperfusion.
Area of Science:
- Neuroscience
- Immunology
- Pathophysiology
Context:
- Cerebral ischemia/reperfusion (I/R) injury is a major cause of stroke.
- Monocyte subsets play a critical role in the inflammatory response following brain injury.
- Understanding the dynamic changes of these subsets is crucial for developing therapeutic strategies.
Purpose:
- To investigate the dynamic changes of Ly6C(high) and Ly6C(low) monocyte subsets after middle cerebral artery occlusion/reperfusion (MCAO/R) in a mouse model.
- To determine the correlation between these monocyte subset changes and cerebral infarct size and neurological deficits.
Summary:
- Ly6C(high) monocyte proportions significantly increased one day after MCAO/R, peaking on day two.
- Compared to sham controls, Ly6C(high) monocytes were elevated at all time points post-MCAO/R.
- Infarct size, assessed by TTC staining, increased markedly by day two.
- A positive correlation was observed between Ly6C(high) monocyte levels, infarct size, and neurological severity scores.
Impact:
- This study highlights the dynamic role of Ly6C(high) monocytes in the acute phase of cerebral I/R injury.
- The findings suggest Ly6C(high) monocytes may serve as a potential biomarker for stroke severity.
- Further research could explore targeting these monocytes for therapeutic interventions to mitigate brain damage.
