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Intraluminal Middle Cerebral Artery Occlusion (MCAO) Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice
Published on: May 8, 2011
Middle cerebral artery occlusion in the mouse by intraluminal suture coated with poly-L-lysine: neurological and
1Cerebral Vascular Disease Research Center, Department of Neurology (D4-5), University of Miami School of Medicine, PO Box 016960, Miami, FL 33101, USA. lbelayev@stroke.med.miami.edu
Abstract:
The present study was conducted to validate a modified method of temporary focal cerebral ischemia in the mouse; neurobehavioral function and histopathological infarction were quantitated following various periods of middle cerebral artery occlusion (MCAo). Male C57BL/6 mice were anesthetized with 3% halothane in a mixture of 30%O2/70%N2O delivered by face mask and were subjected to 30- to 180-min of temporary middle cerebral artery occlusion (MCAo) by an intraluminal suture coated with poly-l-lysine. Twenty-eight of 40 mice showed an initial high-grade neurological deficit (30-min MCAo, n=7; 60-min, n=8; 120-min, n=8; 180-min, n=5) when examined during MCAo; these were used for subsequent study. One day after MCAo, behavioral function was re-evaluated, and brains were perfusion-fixed and infarct volumes were measured. The initial neurological deficit improved at 24 h in mice with 30- or 60-min of prior MCAo but tended to persist in mice with 120- or 180-min insults. Following each duration of ischemia, mice exhibited ipsilateral infarcts. Small, inconsistent predominantly subcortical infarcts were present after 30-min MCAo, while longer occlusion periods gave rise to consistent foci of subcortical infarction involving striatum, septum, thalamus, and hippocampus, as well as areas of frontoparietal cortical infarction. The major advantages of the improved intraluminal MCAo model reported here, incorporating sutures coated with poly-l-lysine, include: a 100% incidence of infarction of predictable location and size in mice having an initial neurological deficit. Periods of 60- to 180-min MCA occlusion in this model yield sufficiently reproducible sequelae to permit the effects of various therapeutic agents on neurological outcome and size of infarction to be readily studied.
Insights
This study validates a modified mouse model for temporary focal cerebral ischemia using middle cerebral artery occlusion (MCAo). The improved method ensures predictable infarcts, aiding stroke research.
Area of Science:
- Neuroscience
- Cerebrovascular Research
- Animal Models
Background:
- Stroke research requires reliable animal models to study disease mechanisms and test therapeutics.
- Temporary focal cerebral ischemia models are crucial for mimicking ischemic stroke conditions.
- Existing models may lack consistency in infarct size and location, hindering research reproducibility.
Purpose of the Study:
- To validate a modified intraluminal suture method for temporary focal cerebral ischemia in mice.
- To assess neurobehavioral deficits and histopathological infarcts following varying durations of middle cerebral artery occlusion (MCAo).
- To establish a reproducible stroke model for evaluating therapeutic interventions.
Main Methods:
- Male C57BL/6 mice underwent temporary middle cerebral artery occlusion (MCAo) for 30 to 180 minutes using a poly-l-lysine coated suture.
- Neurobehavioral deficits were assessed during MCAo and 24 hours post-occlusion.
- Infarct volumes were measured histopathologically after perfusion fixation.
Main Results:
- A modified MCAo technique with a poly-l-lysine coated suture produced a 100% incidence of infarction with predictable location and size.
- Initial neurological deficits improved after 30- or 60-minute MCAo but persisted after 120- or 180-minute occlusions.
- Longer occlusion periods (60-180 minutes) resulted in consistent subcortical and cortical infarcts, suitable for therapeutic studies.
Conclusions:
- The modified intraluminal MCAo model provides a reliable and reproducible method for inducing focal cerebral ischemia in mice.
- This validated model facilitates the study of neurobehavioral outcomes and infarct development.
- The model's reproducibility supports its utility in preclinical stroke research and drug development.
