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Isolation and Flow Cytometric Analysis of Immune Cells from the Ischemic Mouse Brain
Published on: February 12, 2016
Inflammatory reaction after focal cerebral ischemia in mouse
Zhi-Qiu Wang1, Xian-Cheng Chen, Guo-Yuan Yang
1Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai 200040, China. zhiqiuwang@yahoo.com
Background:
In response to the inflammatory reaction, circulating leukocytes aggregate and adhere to the endothelial cells and eventually pervade into tissues, resulting in cell damage. This study was to detect the inflammatory reactions in mouse focal cerebral ischemia and their distinct characteristics in the ischemic basal ganglia and surrounding cortex.
Methods:
Mice were subjected to permanent occlusion of the left middle cerebral artery (MCAO) by introducing a suture for 2 to 120 hours. The expression of intercellular adhesion molecule 1 (ICAM-1) and Mac-1 was determined immunohistochemically. The myeloperoxidase (MPO) activity of the ischemic regions was measured.
Results:
Four hours after MCAO, the number of ICAM-1 positive vessels in the ischemic basal ganglia increased (9.2 +/- 2.8 per mm(2)), peaked at 48 hours (29.6 +/- 4.8 per mm(2)), and decreased after 72 hours. In the ischemic cortex, the number increased rapidly 4 hours after MCAO (19.4 +/- 6.1 per mm(2)), peaked at 48 hours (44.4 +/- 16.8 per mm(2)), and declined after 72 hours. Mac-1 positive cells were seen in the ischemic basal ganglia (3.4 +/- 1.2 per mm(2)) 12 hours after MCAO, peaked after 48 hours (20.2 +/- 6.3 per mm(2)), and decreased after 72 hours. In the ischemic cortex, however, the number increased 4 hours after MCAO (4.3 +/- 1.7 per mm(2)), peaked after 48 hours (20.9 +/- 8.4 per mm(2)), and remained high at 120 hours. The MPO activity increased in the ischemic basal ganglia 12 hours after MCAO (0.111 +/- 0.023 U/g), peaked after 24 hours (0.194 +/- 0.059 U/g), and decreased after 72 hours. In the ischemic cortex, the MPO activity increased 12 hours after MCAO (0.110 +/- 0.032 U/g), peaked after 24 hours (0.210 +/- 0.067 U/g), and remained elevated at 120 hours.
Conclusions:
The increased expression of ICAM-1 in the ischemic brain of mouse in the early phase of MCAO followed by the over-expression of Mac-1 and the increased MPO activity suggests that focal ischemia leads to early onset of inflammation. The inflammatory response is more persistent and intensive in the ischemic cortex than in the ischemic basal ganglia.
Insights
Inflammation in mouse focal cerebral ischemia involves increased intercellular adhesion molecule 1 (ICAM-1) and Mac-1 expression, along with myeloperoxidase (MPO) activity. The inflammatory response is more prolonged and intense in the ischemic cortex compared to the basal ganglia.
Area of Science:
- Neuroscience
- Immunology
- Pathology
Background:
- Focal cerebral ischemia triggers an inflammatory response involving leukocyte adhesion and tissue damage.
- Understanding the distinct inflammatory characteristics in different brain regions is crucial for therapeutic development.
Purpose of the Study:
- To investigate the inflammatory reactions in mouse focal cerebral ischemia.
- To differentiate the inflammatory characteristics in the ischemic basal ganglia and surrounding cortex.
Main Methods:
- Mice underwent permanent middle cerebral artery occlusion (MCAO) for 2 to 120 hours.
- Immunohistochemistry was used to assess intercellular adhesion molecule 1 (ICAM-1) and Mac-1 expression.
- Myeloperoxidase (MPO) activity was measured in ischemic regions.
Main Results:
- ICAM-1 expression increased in both regions, peaking at 48 hours post-MCAO, with higher levels in the cortex.
- Mac-1 positive cells and MPO activity showed increased expression and activity in both regions, persisting longer in the cortex.
- The inflammatory response, indicated by Mac-1 and MPO, was more sustained in the ischemic cortex than in the basal ganglia.
Conclusions:
- Focal ischemia induces early-onset inflammation characterized by increased ICAM-1, Mac-1, and MPO activity.
- The inflammatory response in the ischemic cortex is more persistent and intensive than in the basal ganglia.
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