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Updated: Aug 17, 2026
Induction of Ischemic Stroke and Ischemia-reperfusion in Mice Using the Middle Artery Occlusion Technique and Visualization of Infarct Area
Published on: February 2, 2017
Mice deficient in Mac-1 (CD11b/CD18) are less susceptible to cerebral ischemia/reperfusion injury
S G Soriano1, A Coxon, Y F Wang
1Department of Anesthesia, Children's Hospital, Boston, MA 02115, USA. soriano@A1.tch.harvard.edu
Background And Purpose:
Macrophage-1 antigen (Mac-1) (CD11b/CD18), a leukocyte beta2 integrin, facilitates neutrophil adhesion, transendothelial migration, phagocytosis, and respiratory burst, all of which may mediate reperfusion-induced injury to ischemic brain tissue in conditions such as stroke. To determine the role of Mac-1 during ischemia and reperfusion in the brain, we analyzed the effect of transient focal cerebral ischemia in mice genetically engineered with a specific deficiency in Mac-1.
Methods:
Transient focal ischemia/reperfusion was induced by occluding the left middle cerebral artery for 3 hours followed by a 21-hour reperfusion period in Mac-1-deficient (n=12) and wild-type (n=11) mice. Regional cerebral blood flow was determined with a laser-Doppler flowmeter. Brain sections were stained with 2% 2,3,5-triphenyltetrazolium chloride to determine the infarct volume. Neutrophil accumulation was determined by staining the brain sections with dichloroacetate esterase to identify neutrophils.
Results:
Compared with the wild-type cohort, Mac-1-deficient mice had a 26% reduction in infarction volume (P<0.05). This was associated with a 50%, but statistically insignificant, reduction in the number of extravasated neutrophils in the infarcted areas of the brains in the mutant mice. There were no differences in regional cerebral blood flow between the 2 groups.
Conclusions:
Mac-1 deficiency reduces neutrophil infiltration and cerebral cell death after transient focal cerebral ischemia. This finding may be related to a reduction in neutrophil extravasation in Mac-1-deficient mice.
Insights
Macrophage-1 antigen (Mac-1) deficiency significantly reduced brain infarction volume and neutrophil infiltration in mice after stroke. This suggests Mac-1 plays a key role in stroke-related brain injury.
Area of Science:
- Neuroscience
- Immunology
- Cardiovascular Biology
Background:
- Macrophage-1 antigen (Mac-1) is a leukocyte integrin involved in neutrophil adhesion and migration.
- Mac-1 may contribute to reperfusion injury in ischemic brain tissue, such as in stroke.
Purpose of the Study:
- To investigate the role of Mac-1 in the brain during focal cerebral ischemia and reperfusion.
- To analyze the effects of Mac-1 deficiency on stroke outcomes in a mouse model.
Main Methods:
- Transient focal cerebral ischemia was induced in Mac-1-deficient and wild-type mice.
- Infarct volume was measured using 2,3,5-triphenyltetrazolium chloride staining.
- Neutrophil accumulation was quantified via dichloroacetate esterase staining.
Main Results:
- Mac-1-deficient mice exhibited a 26% reduction in infarct volume compared to wild-type mice.
- A trend towards reduced neutrophil extravasation was observed in Mac-1-deficient mice.
- No significant differences in regional cerebral blood flow were detected between groups.
Conclusions:
- Mac-1 deficiency reduces cerebral cell death and neutrophil infiltration following transient focal cerebral ischemia.
- Reduced neutrophil extravasation in Mac-1-deficient mice may underlie the observed neuroprotective effect.
