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A Rat Model of Middle Cerebral Artery Occlusion/Reperfusion Without Damaging the Anatomical Structure of Cerebral Vessels
In This Article
Summary
The current protocol describes a rat model of middle cerebral artery occlusion/reperfusion that preserves the anatomical structure of cerebral vessels without causing damage.
Abstract
Ischemic stroke stands as the primary cause of long-term disability and mortality among adults worldwide. Animal models of ischemic stroke have significantly contributed to our understanding of its pathological mechanisms and the development of potential treatments. Presently, there are two common methods involving filament (endovascular suture) techniques to induce animal models of cerebral ischemia. However, these methods have inherent limitations, such as reduced blood perfusion to the brain, damage to the external carotid artery system, impaired food and/or water intake, and sensory dysfunction of the face. This article introduces a new method for inducing a rat ischemic stroke model without compromising the cerebral vascular anatomy. In this study, the common carotid artery (CCA) of Sprague-Dawley rats was exposed, and an incision was made. A filament was then inserted through the incision into the internal carotid artery to occlude the middle cerebral artery. After 1.5 h of induced ischemia, the occluding filament was fully removed from both the internal carotid artery and the CCA. The incision in the CCA was subsequently sutured using 11-0 microsurgical sutures under a microscope (magnification 4x). Through the utilization of microsurgical techniques to repair the CCA, this study successfully developed a unique method to induce an ischemic stroke model in rats while preserving the anatomical integrity of cerebral blood vessels.
Introduction
Stroke, the leading cause of death and long-term neurological dysfunction in adults worldwide, encompass various types, with ischemic stroke accounting for approximately 81.9% of cases, while cerebral hemorrhage and subarachnoid hemorrhage account for 14.9% and 3.1%, respectively1. Ischemic stroke often results from atherosclerosis or occlusion of the middle cerebral artery (MCA), leading to local cerebral blood flow reduction and subsequent brain damage. This reduction in nutrient supply to ischemic cells triggers energy depletion, causing membrane integrity loss, cell swelling, and eventual lysis. Additionally, factors like increased intracel....
Protocol
The experimental procedure received approval from the Institutional Animal Care and Use Committee at Yantai University (Approval Number: YTDX20230410) and adhered to the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals. Male Sprague-Dawley rats weighing between 300-350 g were utilized for this study. A comprehensive list of the animals, reagents, and equipment used is provided in the Table of Materials.
1. Animal preparation
Representative Results
The process of microsuture
The filament was withdrawn near the incision of the CCA until the white filament tip was visible (Figure 1A). Micro clamps were then placed on the CCA (Figure 1B). Under a microscope at 4x magnification, the incision of the CCA was sutured using microsurgical sutures in an interrupted pattern (Figure 1C). Subsequently, the microclamps were removed from the CCA, restoring the anatomic.......
Discussion
MCAo is a widely used method to create animal models of stroke, effectively replicating the stroke process observed in patients by restoring blood flow after ischemia10,11,12. Our laboratory has developed an ischemic stroke model that enables reperfusion from the CCA without causing damage to the ECA. Following the induction of ischemia, microsurgical repair of the CCA is performed. Although direct measurement of CCA blood flow .......
Acknowledgements
This work was supported by the Hebei Province Introducing Foreign Intelligence Project in 2023. We thank Lisa Kreiner, PhD, from Liwen Bianji, (Edanz) (www.liwenbianji.cn), for editing the English text of a draft of this manuscript.
....Materials
| Name | Company | Catalog Number | Comments |
| 10% formalin | Beijing Labgic Technology Co.,Ltd. | BL913A | |
| 11-0 microsurgical sutures | Ningbo Medical Suture Needle Co., Ltd. | 3/8 1×5 | |
| 2,3,5-triphenyltetrazolium chloride (TTC) | SIGMA | T8877 | |
| 3-0 silk suture | Ningbo Medical Suture Needle Co., Ltd. | 3-0 | |
| 4% isoflurane | Tianjin Ringpu Bio-Technology Co., Ltd. | 20221102 | |
| 6-0 silk suture | Ningbo Medical Suture Needle Co., Ltd. | 6-0 | |
| 70% ethanol | Shandong Lierkang Medical Technology Co., Ltd. | 20230408 | |
| Filament | Xinong Technologies Co Ltd. | 2838-A5 | |
| Ketamine | Jiangsu Hengrui Pharmaceutical Co., Ltd. | 230613BL | |
| Male Sprague Dawley rats | Jinan Pengyue Experimental Animal Breeding Co., Ltd. | SCXK (Lu) 20190003 | |
| Microclips | Shanghai Jinzhong Medical Device Factory | W40150 | |
| Microscope | ZEISS | S100 OPMI pico | |
| Nursing box | Beijing Fuyi Electrical Appliance Co., Ltd. | FYL-YS-100L | |
| Xylazine | SIGMA | PHR3263 |
References
- Tu, W. J., et al. China stroke surveillance report 2021. Mil Med Res. 10 (1), 33 (2023).
- Wu, L., et al. Targeting oxidative stress and inflammation to prevent ischemia-reperfusion injury. Front in ....
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