We would like to thank Dr. Lei Chen (Icahn School of Medicine at Mount Sinai, NY) who first established this model in our laboratory. Supported in part by HL126559, DA039576, MH098891, MH63022, MH072567, DA027569, and NSC 2015/17/B/NZ7/02985. Dr. Luc Bertrand is supported in part by a postdoctoral fellowship from the American Heart Association (16POST31170002).

所有的程序是由迈阿密机构动物护理和使用委员会（IACUC）的大学按照卫生研究院（NIH）的指导方针的国家机构的批准。需要无菌设备和无菌技术的使用。 1.准备缝合闭塞<ol><li>使用直径0.21毫米的缝合线20之间的小鼠 - 25克0.23毫米至25只小鼠 - 35克体重的。缝线的MCAO程序的类型的选择取决于动物的体重。 </li><li>使用银笔，标记从涂硅尖开始9毫米缝合。这将作为插入长度的导向。 </li></ol> 2.准备手术<ol><li>麻醉小鼠用异氟烷与氧混合，用实验室麻醉系统。在凌晨2商用机器上设置5和氧气流量使用异氟醚（见材料表 ）。动物转移到surgerŸ表面用鼻锥维持麻醉（异氟烷使用1.5设置 - 在2 2.5和氧气流量）。 <ol><li>确保鼠标的呼吸率约为1 - 每秒2呼吸喘气没有。此外，确保动物不会出现晶须刺激的反应和踏板反射（趾捏）。监控呼吸率和精力的手术过程中，至少每5分钟。 </li></ol></li><li>放眼用润滑剂上每只眼睛一滴使用无菌拭子，以防止它们在手术过程中进行干燥。可替代地，从一个无菌药物管适用眼药膏的眼球。 </li><li>翻转在它的后面的动物和刮胡子切口区域。之后，彻底消毒用70％的乙醇，接着洗必泰，并用70％乙醇的最终拭子的外科手术区域。放置在一个温暖的手术表面的动物立体显微镜下。 </li></ol> 3.颈总动脉夹层和内部/ EXTERNAL分支<ol><li>用手术剪和镊子，在颈部执行浅中线切口，从乳房骨上面的下颚下面（大约3 - 第4厘米）。手术单应放在周围的手术区域，以防止仪器从与非无菌表面接触。外科被单在当前视频省略以有助于成膜。 </li><li>使用镊子，仔细分离脂肪和结缔组织暴露气管。组织应该轻松，自然分开双方。 </li><li>放置在小鼠的颈背部枕头（圆形物体，约0.5厘米，直径），以延伸颈部，进一步暴露手术的区域。 </li><li>打开使用一个组织牵开器或钩切口。 </li><li>上气管的动物的左侧，小心tweeze相距结缔组织以暴露左颈总动脉（CCA）。要小心，不要损坏与CCA捆绑的神经和主脉。 </li><li>继续露出CCA，小心翼翼地从底层组织分离，并露出顶部"Y"分支内部的脑动脉（ICA）和外部脑动脉（ECA）的。要卸下CCA帮助，将CCA下弯钳刺穿结缔组织，然后慢慢让他们打开。 </li></ol> 4.制备CCA为缺血缝线插针<ol><li>插入的长度约4厘米的CCA下尼龙缝线的3段。确保CCA不扭曲，因为这将极大地缝合线的插入变得复杂。 </li><li>在最低点可能，使用永久结关闭底部缝合。 </li><li>领带就在ICA下方的顶缝/ ECA分支使用可移动的滑结。 </li><li>配合使用可移动滑结中间缝合，但保持敞开的。它离开足够的空间是重要的，因为不阻碍缝合插入。 </li><li>采用显微春天科幻ssors，在CCA进行底部和中部之间缝合切口。执行0.2毫米的切口接近底部缝合。 </li></ol> 5.大脑中动脉闭塞<ol><li>使用镊子，将缝线缺血进入CCA切口，引导其到顶部。 4.5步后迅速执行此步骤，以防止CCA开放的凝血和关闭。在缝合插入受阻的情况下，用钳提示重新打开切口。 </li><li>轻轻用滑结，以限制其周围血流阕中间缝合在MCAO缝合硅氧烷部分，但足够宽松以允许其自由移动。 </li><li>仔细撤消顶部缝合，确保了缺血缝合不滑出。 </li><li>通过几毫米的插入中的ICA的MCAO缝合，然后重新关闭的顶部缝合，以相同的方式如在步骤5.2中描述。 </li><li>引导缺血缝合到闭塞区域。成功插入指示从CCA切口少量血液回流的，而且银9毫米明显的标志是位于CCA切口和ICA / ECA分叉之间。可使用的监测方法如激光多普勒血流监测系统15，16来获得成功闭塞进一步确认。 注:在血流90％A下降至在闭塞区域侧中部脑区域表示成功闭塞。 </li><li>成功闭塞后，紧紧牵制中部和顶部缝合。如果执行假的插入，不重合缝合而是立即跳到步骤7.3。 </li></ol> 6.切口关闭和术后护理<ol><li>塔克在缝线到切口区域。 </li><li>删除拉钩或组织挂钩和枕头。 </li><li>使用无菌盐水和棉签清洁缝合区。 </li><li>关闭使用尼龙线/针和镊子的切口。 </li> &lt;利&gt;管理抗炎（ 例如 ，卡洛芬10毫克/千克，SC）和镇痛药（ 例如 ，丁丙诺啡0.1毫克/千克SC，每天两次）的药物，以减轻术后不适。放置在老鼠放在一个加热垫一个笼子里，以防止低温和给水软化食品广告阴唇访问。 </li><li>监测动物的恢复（5 - 10分钟）和中风的迹象。他们可以从轻度麻痹侧各不相同，盘旋对侧侧翼和滚动的剧烈收缩。几个评价观点中列出了讨论。显示呼吸窘迫或严重癫痫发作的迹象动物应实施安乐死。 </li><li>取决于缺血再灌注协议，从30分钟到120分钟或更离开代替MCAO缝合。减少阻塞时间，可以防止死亡。 </li><li>对于永久性闭塞，留在原地的缝合缺血24小时，但显著的死亡率是可以预期的。这段时间过去后，继续STE第7页。 <ol><li>如果使用永久性闭塞模型，养老鼠笼，直到终点。当时，去除缝线（步骤7.1到7.8）。这个过程也可以安乐死之后进行。 </li></ol></li></ol> 7.再灌注<ol><li>再次麻醉动物（按照2.1的说明），并删除创伤闭合缝线。 </li><li>使用镊子和组织分离器，重新打开切口暴露CCA。 </li><li>小心取出顶部缝合，直到涂硅部分位于中间结对缺血缝合轻轻一拉。 </li><li>重做在缝合顶端结防止血液流入通过缝合（它不必是一滑结）。 </li><li>仔细撤消中间结，拉动缝线过去结顶，但保持了CCA内。 </li><li>盖紧结顶阻断动脉血流。 </li><li>拉出完全缺血缝合然后拧紧中间结。 </li><li>在MCAO缝合可以是重复使用几次。在每次使用后，仔细清洁缝合在70％乙醇以除去使用无菌纱布的污染物（ 例如 ，血液或组织）。之后，放置在消毒袋缝合和消毒它。 </li><li>重复步骤6.1 - 6.4和监视动物的恢复。 </li></ol> 8.组织分析<ol><li>为了监测每搏输出量，剖析了大脑。 <ol><li>使用两步骤过程安乐死鼠标。首先，暴露动物异氟醚过量，直到呼吸停止。对于TTC染色，立即斩首鼠标进行脑部提取。脑切片（ 如用于免疫分析），通过前一个斩首心脏穿刺灌注执行。 </li><li>切断在颅底头和切断从颈部开口的头骨顶部的头皮到眼睛之间。与颅骨外露，切沿两侧从颅底开始开骨到眼窝。使沿着头骨的一部分，广本晋级。要小心，不要达到用剪刀剪，以免损坏大脑太深。 </li><li>此后，切两眼窝之间的骨头。 </li></ol></li><li>使用镊子，抬起头颅顶部，露出大脑。应该只有一个小的阻力。如果不轻松举起，头骨切口可能不完整。撬出轻轻用钝弯钳大脑。大脑仍可能由几个神经被连接。继续慢慢地，沿途移除附件。 </li><li>放置新鲜脑在1毫米大脑基质，加入一对的PBS滴，并沿着用剃刀刀片或低温恒温器叶片基质切片。 </li><li>脑转移到100mm培养皿，并添加染色溶液（2％2,3,5-氯化三苯四唑溶解在PBS [TTC]）以覆盖培养皿的底部。使用两个镊子，独立的所有脑切片（正面朝下），并将其放置，以便在培养皿的底部。 注:染色会变成活组织的红色，与非活组织剩余的白色。温热该溶液至37℃将加速染色。中风面积（白色）的症状可以迅速视为90分钟后中风且持续超过7天可见。成像应做连同一把尺子，使每搏输出量计算。 </li><li>过程解剖大脑用于切片，免疫染色，蛋白质分离，或各种其他程序17，18，19。 </li></ol>

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Authors have no competing or conflicts of interest.

所描述的MCAO方法的成功应用是高度依赖于脑血流解剖学的理解。由于缝合线的正确位置是难以辨别，由于缺乏直接的视觉线索，反复练习很重要，将其用于调查研究之前掌握的过程。行程容积应分析，以确保一致的结果。增加了一个激光多普勒系统可以帮助确定血流的成功闭塞和应定期使用，以确保过程正确进行。路由MCAO缝合到闭塞区域可以通过操纵动脉来促进。为了帮助在缝线引导至MCA，一?...

心血管疾病的研究中，如中风，依赖于使用的体内模型。理解局部缺血，药物毒性，和/或治疗的可能含义，有必要使用该疾病，从而使治疗组之间的比较研究的一个合适的，标准化的，可靠的和可再现的模型。在这个手稿中，我们使用的是老鼠，给予了大量的转基因小鼠和标准化的评估模型的有效性。耙分数来评估以下的实验缺血性卒中及以下恢复运动和行为障碍已经开发的1，2。 几个缺血性中风型号可供选择，如完整的全球脑缺血，不完全性缺血，多灶性脑缺血和脑缺血。后一组也是中风患者最常见的种类。大多数前夕NTS由栓塞或血栓闭塞的形成在或接近大脑中动脉（MCA）发起。鉴于这些参数，紧紧提出的模型模拟人类中风的疾病的病因，使获得高度相关的3个结果。尽管如此，从动物模型在人类疾病治疗的发现的翻译已被证明是具有挑战性的。截至目前，只有使用溶栓组织型纤溶酶原激活物已被批准用于治疗急性缺血性脑卒中4。 间在小鼠脑缺血，大脑后循环中风模型和脑静脉血栓形成模型的模型是高度侵入性，减少它们的适用性，并限制能够进行分析的范围。然而，其他技术如栓塞模型，photothrombosis模型，内皮素-1诱导的中风模型，和管腔内缝合大脑中动脉闭塞（MCAO）模型，可用于使用没有这些限制。在缺血模型在本协议中所述的技术。它提供了诱导可以容易地再灌注，并在高通量的方式进行局部脑缺血的可靠方法。有两种方法这一模式，即玉米，龙格和小泉的方法。它们在闭塞缝合插入脉管的方式略有不同。在玉米-隆加技术，缝线经由外颈动脉5插入。这里提出的技术是从在其中封闭缝合经由颈总动脉6插入小泉方法改性。 该MCAO模型已成功地应用于评估缺血性中风期间发生的不同事件。以下再灌注，脑水肿可以与血脑屏障的击穿可以观察到沿。山顶神经元死亡通常在24小时观察;然而，将其重新后7天7转动到基线水平。在人类中，性别和年龄的确定行程结果时，这也将在小鼠和大鼠8，9，10观察的重要变量。若干出版物已经使用了MCAO模型以证明处理效率11，12，13，14。

<table><tbody><tr><td>MCAO suture 0.23 mm</td><td>Doccol</td><td>702345PK5Re</td><td></td></tr><tr><td>MCAO suture 0.21 mm</td><td>Doccol</td><td>702145PK5Re</td><td></td></tr><tr><td>Silver pen</td><td>staples</td><td>503205</td><td></td></tr><tr><td>Anesthesia machine</td><td>Vetequip</td><td>901806</td><td></td></tr><tr><td>Surgical scissors</td><td>Fine science tool</td><td>14558-09</td><td></td></tr><tr><td>Surgical forceps straight tip</td><td>Fine science tool</td><td>00108-11</td><td></td></tr><tr><td>Surgical forceps angled tip</td><td>Fine science tool</td><td>00109-11</td><td></td></tr><tr><td>Spring scissors</td><td>Fine science tool</td><td>15000-08</td><td></td></tr><tr><td>Nylon suture</td><td>Braintree scintific</td><td>SUT-S 104</td><td></td></tr><tr><td>Closing suture</td><td>VWR</td><td>95057-036</td><td></td></tr><tr><td>Isoflurane</td><td>Piramal</td><td></td><td></td></tr><tr><td>2,3,5-Triphenyltetrazolium chloride</td><td>FisherSci</td><td>50-121-8005</td><td></td></tr><tr><td>Brain block</td><td>Braintree scintific</td><td>BS-A 5000C</td><td></td></tr><tr><td>Cryostat blade</td><td>VWR</td><td>89202-606</td><td></td></tr><tr><td>Optional:</td><td></td><td></td><td></td></tr><tr><td>Periflux Laser doppler system</td><td>Perimed</td><td>Periflux 5000</td><td></td></tr><tr><td>Monitoring unit</td><td>Perimed</td><td>PF 5010 - LDPM</td><td></td></tr></tbody></table>

induction of ischemic stroke and ischemia-reperfusion in mice using the middle artery occlusion technique and visualization of infarct area

Cerebrovascular disease is highly prevalent in the global population and encompasses several types of conditions, including stroke. To study the impact of stroke on tissue injury and to evaluate the effectiveness of therapeutic interventions, several experimental models in a variety of species were developed. They include complete global cerebral ischemia, incomplete global ischemia, focal cerebral ischemia, and multifocal cerebral ischemia. The model described in this protocol is based on the middle cerebral artery occlusion (MCAO) and is related to the focal ischemia category. This technique produces consistent focal ischemia in a strictly defined region of the hemisphere and is less invasive than other methods. The procedure described is performed on mice, given the availability of several genetic variants and the high number of tests standardized for mice to aid in the behavioral and neurodeficit evaluation.

为闭塞缝线的插入路径是表现在图1中 。在MCAO缝合要被路由到闭塞区域，在ICA中分叉。马华的成功闭塞会导致组织损伤，TTC染色可见。 图2呈现从假治疗的动物（ 图2A）和从60分钟缺血缺血再灌注动物染色的图像（染色，在90分钟或24小时后的闭塞， 图2B）。为了确定每搏量，首先计算行程区的每个部分中，使用包括在成像...

Watch this Scientific Journal Video about Induction of Ischemic Stroke and Ischemia-reperfusion in Mice Using the Middle Artery Occlusion Technique and Visualization of Infarct Area at JoVE.com

Induction of Ischemic Stroke and Ischemia-reperfusion in Mice Using the Middle Artery Occlusion Technique and Visualization of Infarct Area

We describe a mouse model of stroke induced by the occlusion of the middle cerebral artery using a silicone coated suture. The protocol can be applied to induce permanent occlusion or a temporary ischemia, followed by reperfusion.

缺血性中风和缺血再灌注小鼠的诱导使用梗塞区的中心动脉闭塞技术和可视化

Cerebrovascular disease is highly prevalent in the global population and encompasses several types of conditions, including stroke. To study the impact of ...

induction-ischemic-stroke-ischemia-reperfusion-mice-using-middle

Research

JoVE Journal

Medicine

16.2K 次观看.  University of Miami. We describe a mouse model of stroke induced by the occlusion of the middle cerebral artery using a silicone coated suture. The protocol can be applied to induce permanent occlusion or a temporary ischemia, followed by reperfusion.

视频: Induction of Ischemic Stroke and Ischemia-reperfusion in Mice Using the Middle Artery Occlusion Technique and Visualization of Infarct Area

Focal Cerebral Ischemia Model by Endovascular Suture Occlusion of the Middle Cerebral Artery in the Rat

Stroke is the leading cause of disability and the third leading cause of death in adults worldwide1. In human stroke, there exists a highly variable clinical state; in the development of animal models of focal ischemia, however, achieving reproducibility of experimentally induced infarct volume is essential. The rat is a widely used animal model for stroke due to its relatively low animal husbandry costs and to the similarity of its cranial circulation to that of humans2,3. In humans, the middle cerebral artery (MCA) is most commonly affected in stroke syndromes and multiple methods of MCA occlusion (MCAO) have been described to mimic this clinical syndrome in animal models. Because recanalization commonly occurs following an acute stroke in the human, reperfusion after a period of occlusion has been included in many of these models. In this video, we demonstrate the transient endovascular suture MCAO model in the spontaneously hypertensive rat (SHR). A filament with a silicon tip coating is placed intraluminally at the MCA origin for 60 minutes, followed by reperfusion. Note that the optimal occlusion period may vary in other rat strains, such as Wistar or Sprague-Dawley. Several behavioral indicators of stroke in the rat are shown. Focal ischemia is confirmed using T2-weighted magnetic resonance images and by staining brain sections with 2,3,5-triphenyltetrazolium chloride (TTC) 24 hours after MCAO.

Surgical induction of ischemic brain damage in the rat is a widely used model for stroke research. Here we demonstrate the induction of focal cerebral ischemia by occlusion of the middle cerebral artery. Visualization of the resulting infarct by histological staining and magnetic resonance imaging is also shown.

焦距脑缺血模型大鼠大脑中动脉的血管内缝合闭塞

Stroke is the leading cause of disability and the third leading cause of death in adults worldwide1. In human stroke, there exists a highly variable ...

科研

神经科学

Mouse Model of Middle Cerebral Artery Occlusion

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the brain (ischemic stroke) 2. Since most ischemic strokes (~80%) occur in the territory of middle cerebral artery (MCA) 3, many animal stroke models that have been developed have focused on this artery. The intraluminal monofilament model of middle cerebral artery occlusion (MCAO) involves the insertion of a surgical filament into the external carotid artery and threading it forward into the internal carotid artery (ICA) until the tip occludes the origin of the MCA, resulting in a cessation of blood flow and subsequent brain infarction in the MCA territory 4. The technique can be used to model permanent or transient occlusion 5. If the suture is removed after a certain interval (30 min, 1 h, or 2 h), reperfusion is achieved (transient MCAO); if the filament is left in place (24 h) the procedure is suitable as a model of permanent MCAO. This technique does not require craniectomy, a neurosurgical procedure to remove a portion of skull, which may affect intracranial pressure and temperature 6. It has become the most frequently used method to mimic permanent and transient focal cerebral ischemia in rats and mice 7,8. To evaluate the extent of cerebral infarction, we stain brain slices with 2,3,5-triphenyltetrazolium chloride (TTC) to identify ischemic brain tissue 9. In this video, we demonstrate the MCAO method and the determination of infarct size by TTC staining.

We demonstrate in the video a method for producing a middle cerebral artery occlusion in adult mice using an intraluminal monofilament. We also show how to evaluate the extent of cerebral infarction by 2,3,5-triphenyltetrazolium chloride (TTC) staining.

大脑中动脉闭塞的小鼠模型

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the ...

医学

Intraluminal Middle Cerebral Artery Occlusion (MCAO) Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice

Stroke is the third leading cause of death and the leading cause of disability in the world, with an estimated cost of near $70 billion 
in the United States in 20091,2. The intraluminal middle cerebral artery occlusion (MCAO) model was developed by Koizumi4 in 1986 to simulate this 
impactful human pathology in the rat. A modification of the MCAO method was later presented by Longa3. Both techniques have been widely used to identify 
molecular mechanisms of brain injury resulting from ischemic stroke and potential therapeutic modalities5. This relatively noninvasive method in rats has been 
extended to use in mice to take advantage of transgenic and knockout strains6,7. To model focal cerebral ischemia, an intraluminal suture is advanced via 
the internal carotid artery to occlude the base of the MCA. Retracting the suture after a specified period of time mimics spontaneous reperfusion, but the 
suture can also be permanently retained. This video will be demonstrating the two major approaches for performing intraluminal MCAO procedure in mice in a 
stepwise fashion, as well as providing insights for potential drawbacks and pitfalls. The ischemic brain tissue will subsequently be stained by 
2,3,5-triphenyltetrazolium chloride (TTC) to evaluate the extent of cerebral infarction8.

Intraluminal Middle Cerebral Artery Occlusion MCAO Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice

The intraluminal middle cerebral artery occlusion (MCAO) model is the most frequent used model among experimental ischemic stroke models. Here we will demonstrate the entire model in detail with the guide of Laser Doppler flowmetry, and its representative results.

腔内中东大脑中动脉闭塞（MCAO）缺血性中风模型在小鼠体内的激光多普勒血流指导

Stroke is the third leading cause of death and the leading cause of disability in the world, with an estimated cost of near $70 billion 
in the United ...

The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

Focal cerebral ischemia is among the most common type of stroke seen in patients. Due to the clinical significance there has been a prolonged effort to develop suitable animal models to study the events that unfold during ischemic insult. These techniques include transient or permanent, focal or global ischemia models using many different animal models, with the most common being rodents.
The permanent MCA ligation method which is also referred as pMCAo in the literature is used extensively as a focal ischemia model in rodents 1-6. This method was originally described for rats by Tamura et al. in 1981 7. In this protocol a craniotomy was used to access the MCA and the proximal regions were occluded by electrocoagulation. The infarcts involve mostly cortical and sometimes striatal regions depending on the location of the occlusion. This technique is now well established and used in many laboratories 8-13. Early use of this technique led to the definition and description of &#x201C;infarct core&#x201D; and &#x201C;penumbra&#x201D; 14-16, and it is often used to evaluate potential neuroprotective compounds 10, 12, 13, 17. Although the initial studies were performed in rats, permanent MCA ligation has been used successfully in mice with slight modifications 18-20 .
This model yields reproducible infarcts and increased post-survival rates. Approximately 80% of the ischemic strokes in humans happen in the MCA area 21 and thus this is a very relevant model for stroke studies. Currently, there is a paucity of effective treatments available to stroke patients, and thus there is a need for good models to test potential pharmacological compounds and evaluate physiological outcomes. This method can also be used for studying intracellular hypoxia response mechanisms in vivo. 
Here, we present the MCA ligation surgery in a C57/BL6 mouse. We describe the pre-surgical preparation, MCA ligation surgery and 2,3,5 Triphenyltetrazolium chloride (TTC) staining for quantification of infarct volumes.

Middle cerebral artery (MCA) ligation is a technique to study focal cerebral ischemia in animal models. In this method, the middle cerebral artery is exposed by craniotomy and ligated by cauterization. This method gives highly reproducible infarct volumes and increased post-operative survival rates compared to other methods available.

在小鼠永久性大脑中动脉结扎术中的应用

Focal cerebral ischemia is among the most common type of stroke seen in patients. Due to the clinical significance there has been a prolonged effort to ...

Mouse Model of Intraluminal MCAO: Cerebral Infarct Evaluation by Cresyl Violet Staining

Stroke is the third cause of mortality and the leading cause of disability in the World. Ischemic stroke accounts for approximately 80% of all strokes. However, the thrombolytic tissue plasminogen activator (tPA) is the only treatment of acute ischemic stroke that exists. This led researchers to develop several ischemic stroke models in a variety of species. Two major types of rodent models have been developed: models of global cerebral ischemia or focal cerebral ischemia. To mimic ischemic stroke in patients, in whom approximately 80% thrombotic or embolic strokes occur in the territory of the middle cerebral artery (MCA), the intraluminal middle cerebral artery occlusion (MCAO) model is quite relevant for stroke studies. This model was first developed in rats by Koizumi et al. in 1986 1. Because of the ease of genetic manipulation in mice, these models have also been developed in this species 2-3.
	Herein, we present the transient MCA occlusion procedure in C57/Bl6 mice. Previous studies have reported that physical properties of the occluder such as tip diameter, length, shape, and flexibility are critical for the reproducibility of the infarct volume 4. Herein, a commercial silicon coated monofilaments (Doccol Corporation) have been used. Another great advantage is that this monofilament reduces the risk to induce subarachnoid hemorrhages. Using the Zeiss stereo-microscope Stemi 2000, the silicon coated monofilament was introduced into the internal carotid artery (ICA) via a cut in the external carotid artery (ECA) until the monofilament occludes the base of the MCA. Blood flow was restored 1 hour later by removal of the monofilament to mimic the restoration of blood flow after lysis of a thromboembolic clot in humans. The extent of cerebral infarct may be evaluated first by a neurologic score and by the measurement of the infarct volume. Ischemic mice were thus analyzed for their neurologic score at different post-reperfusion times. To evaluate the infarct volume, staining with 2,3,5-triphenyltetrazolium chloride (TTC) was usually performed. Herein, we used cresyl violet staining since it offers the opportunity to test many critical markers by immunohistochemistry. In this video, we report the MCAO procedure; neurological scores and the evaluation of the infarct volume by cresyl violet staining.

The intraluminal middle cerebral occlusion model in mice is herein presented. The extent of cerebral infarct is evaluated by a neurologic score and cresyl violet staining, an alternative staining to TTC, offering the great advantage to test in parallel many interest markers.

小鼠模型的腔内MCAO脑梗塞评价甲酚紫染色

Stroke is the third  cause of mortality and the leading cause of disability in the World. Ischemic  stroke accounts for approximately 80% of all strokes. ...

Surgical Approach for Middle Cerebral Artery Occlusion and Reperfusion Induced Stroke in Mice

Stroke is a leading cause of death worldwide and continues to be one of the major causes of long-term adult disabilities. About 87% of strokes are ischemic in origin and occur in the territory of the middle cerebral artery (MCA). Currently the only Food and Drug Administration (FDA) approved drug for the treatment of this devastating disease is tissue plasminogen activator (tPA). However, tPA has a small therapeutic window for administration (3 - 6 hr), and is only effective in 4% of the patients who actually receive it. Current research focuses on understanding the pathophysiology of stroke in order to find potential therapeutic targets. Thus, reliable models are crucial, and the MCA occlusion (MCAo) model (also termed the intraluminal filament or suture model) is deemed to be the most clinically relevant surgical model of ischemic stroke, and is fairly non-invasive and easily reproducible. Typically the MCAo model is used with rodents, especially with mice due to all the genetic variations available for this species. Here we describe (and present in the video) how to successfully perform the MCAo model (with reperfusion) in mice to generate reliable and reproducible data.

In order to understand the pathophysiology of stroke, it is important to use reliable models. This paper will describe one of the most frequently used stroke models in mice, termed the middle cerebral artery occlusion (MCAo) model (also termed the intraluminal filament or suture model) with reperfusion.

对大脑中动脉闭塞再灌注损伤诱发中风小鼠的手术方法

Stroke is a leading cause of death worldwide and continues to be one of the major causes of long-term adult disabilities. About 87% of strokes are ...

Middle Cerebral Artery Occlusion Allowing Reperfusion via Common Carotid Artery Repair in Mice

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue plasminogen activator (tPA) as an approved drug treatment to target ischemic strokes. Current research in the field of ischemic stroke focuses on better understanding the pathophysiology of stroke, to develop and investigate novel pharmaceutical targets. Reliable experimental stroke models are crucial for the progression of potential treatments. The middle cerebral artery occlusion (MCAO) model is clinically relevant and the most frequently used surgical model of ischemic stroke in rodents. However, the outcomes of this model, such as lesion volume, are associated with high levels of variability, particularly in mice. The alternative MCAO model described here allows the reperfusion of the common carotid artery (CCA) and the increased perfusion of the middle cerebral artery (MCA) territory, using a tissue pad with fibrinogen-based sealant to repair the vessel, and the improved welfare of the mice by avoiding external carotid artery (ECA) ligation. This reduces the reliance on the Circle of Willis, which is known to be highly anatomically variable in mice. Representative data show that using this alternative surgical approach decreases the variability in lesion volumes between the traditional MCAO approach and the alternative approach described here.

Intraluminal filament occlusion of the middle cerebral artery is the most frequently used in vivo model of experimental stroke in rodents. An alternative surgical approach to allow common carotid artery repair is performed here, which allows the reperfusion of the common carotid artery and a full reperfusion to the middle cerebral artery territory.

颅内动脉闭塞允许经普通颈动脉修复再灌注的研究

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue ...

Two-vessel Occlusion Mouse Model of Cerebral Ischemia-reperfusion

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse model is useful for investigating the pathophysiology of cerebral ischemia-reperfusion and determining potential therapeutic strategies for patients with stroke. Variations in the anatomy of the circle of Willis of C57BL/6 mice affects their infarct volume after cerebral-ischemia-induced injury. Studies have indicated that distal MCA occlusion (MCAO) can overcome this problem and result in a stable infarct size. In this study, we establish a two-vessel occlusion mouse model of cerebral ischemia-reperfusion through the interruption of the blood flow to the right MCA. We distally ligate the right MCA and right common carotid artery (CCA) and restore blood flow after a certain period of ischemia. This ischemia-reperfusion injury induces an infarct of stable size and a behavioral deficit. Peripheral immune cells infiltrate the ischemic brain within the 24 h infiltration period. Additionally, the neuronal loss in the cortical area is less for a longer reperfusion duration. Therefore, this two-vessel occlusion model is suitable for investigating the immune response and neuronal recovery during the reperfusion period after cerebral ischemia.

A mouse model of cerebral ischemia-reperfusion is established to investigate the pathophysiology of stroke. We distally ligate the right middle cerebral artery and right common carotid artery and restore blood flow after 10 or 40 min of ischemia.

脑缺血再灌注的双血管闭塞小鼠模型

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse ...

大脑中动脉闭塞的短暂性脑缺血模型

Transient Middle Cerebral Artery Occlusion Model of Stroke

Stroke stands as a major cause of death or chronic disability globally. Nevertheless, existing optimal treatments are limited to reperfusion therapies during the acute phase of ischemic stroke. To gain insights into stroke physiopathology and develop innovative therapeutic approaches, in vivo rodent models of stroke play a fundamental role. The availability of genetically modified animals has particularly propelled the use of mice as experimental stroke models.
In stroke patients, occlusion of the middle cerebral artery (MCA) is a common occurrence. Consequently, the most prevalent experimental model involves intraluminal occlusion of the MCA, a minimally invasive technique that doesn't require craniectomy. This procedure involves inserting a monofilament through the external carotid artery (ECA) and advancing it through the internal carotid artery (ICA) until it reaches the branching point of the MCA. After a 45 min arterial occlusion, the monofilament is removed to allow reperfusion. Throughout the process, cerebral blood flow is monitored to confirm the reduction during occlusion and subsequent recovery upon reperfusion. Neurological and tissue outcomes are evaluated using behavioral tests and magnetic resonance imaging (MRI) studies.

作者聚焦：通过大脑中动脉闭塞模型评估缺血性卒中损伤 

This protocol describes the model of transient focal cerebral ischemia in mice through intraluminal occlusion of the middle cerebral artery. Additionally, examples of outcome assessment are shown using magnetic resonance imaging and behavioral tests.

脑卒中短暂性中动脉闭塞模型

Stroke stands as a major cause of death or chronic disability globally. Nevertheless, existing optimal treatments are limited to reperfusion therapies ...

缺血性中风和缺血再灌注小鼠的诱导使用梗塞区的中心动脉闭塞技术和可视化

摘要

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此视频中的章节

焦距脑缺血模型大鼠大脑中动脉的血管内缝合闭塞

大脑中动脉闭塞的小鼠模型

腔内中东大脑中动脉闭塞（MCAO）缺血性中风模型在小鼠体内的激光多普勒血流指导

在小鼠永久性大脑中动脉结扎术中的应用

小鼠模型的腔内MCAO脑梗塞评价甲酚紫染色

对大脑中动脉闭塞再灌注损伤诱发中风小鼠的手术方法

颅内动脉闭塞允许经普通颈动脉修复再灌注的研究

脑缺血再灌注的双血管闭塞小鼠模型

脑卒中短暂性中动脉闭塞模型

脑卒中短暂性中动脉闭塞模型