Stroke Modeling and Assessment in Mice Using Internal Carotid Artery Occlusion

The internal carotid artery occlusion (ICAO) model in mice replicates a clinically relevant subtype of stroke (mild to moderate ischemic stroke). The ICAO model represents a significant advancement in preclinical stroke research, providing a more accurate representation of human strokes caused by internal carotid artery occlusion. This model has facilitated novel insights into epigenetic modifications following stroke, specifically the dynamics of histone lysine methylation and demethylation, which are crucial in ischemia-induced brain damage and recovery. In contrast to the widely used middle cerebral artery occlusion (MCAO) model, which primarily induces extensive cortical damage, the ICAO model more precisely mimics the striatal and hippocampal injury seen in these stroke cases. Here, we describe the establishment and utilization of the ICAO model in adult CD1 mice, highlighting its reliability and reproducibility in inducing mild to moderate ischemic injury. Our method involves a temporary occlusion of the internal carotid artery for 90 min, followed by reperfusion, leading to localized neural damage. This article details the surgical procedure for inducing ICAO in mice, followed by methods for characterizing the resulting ischemia. These methods include laser doppler perfusion imaging and neurobehavioral assessments, such as neurological deficit scoring and motor function tests. Additionally, the model also emphasizes the importance of considering sex-specific differences in the response to ICAO. This model's ability to yield reproducible and localized neural damage makes it a valuable tool for studying stroke pathophysiology and evaluating potential therapeutic interventions. © 2025 Wiley Periodicals LLC. Basic Protocol 1: Detailed surgical procedure for inducing internal carotid artery occlusion Basic Protocol 2: Laser doppler perfusion imaging Basic protocol 3: Neurobehavioral assessments.