VP is a participant in the Charit&#233; Clinical Scientist Program, funded by the Charit&#233; - Universit&#228;tsmedizin Berlin and the Berlin Institute of Health. TB is an SNSF PostDoc Mobility fellow. The authors receive grant support from EinsteinStiftung/A-2012-153 to PV.

伦理声明:涉及动物受试者实验是根据由Landesamt附耳GESUNDHEIT UND Soziales，柏林，德国（G0298 / 13）和ARRIVE标准规定的准则和规定执行，如适用。在这项研究中，10至12周龄的雄性C57BL / 6J小鼠。 1.横向慢性颅窗口准备<ol><li>用皮下注射氯胺酮（90毫克/千克）和赛拉嗪（10毫克/千克）的执行麻醉。测试与痛觉刺激足够的镇静作用。 </li><li>消毒手术器械和手术区域用70％的乙醇。 </li><li>从颈部用啮齿动物剃须刀的眼睛中取出头皮。 </li><li>在立体定位框架固定的头部。 </li><li>使用上双眼泛醇眼膏，以避免脱水。 </li><li>清洁手术区，以去除所有毛发和74.1％乙醇的3层和10％的2-心得消毒它。 </li><li>执行中线从颈部到用手术刀眼睛切口。 </li><li>跨度皮瓣4隆起缝线。 </li><li>通过用解剖刀到颞肌开始点刮它关闭在左半球小心取出骨膜。 注意:此制剂也用于创建牙科胶，其固定在盖玻璃良好的粘合面积。 </li><li>通过在使用microdrill骨瓣的周缘变薄颅骨执行与直径4mm的额顶叶开颅。应用生理盐水，同时钻探，以避免热损伤。 </li><li>提升骨瓣与插管并使用microforceps将其删除。 </li><li>用生理盐水仔细冲洗广泛。 </li><li>混合所述牙科胶，直到它具有适当的稠度和不流体（ 即，胶不应产生线程了）。将其放在周围的开颅骨头。 </li><li>放置一个玻璃盖的直径在准备好的胶水6毫米，次修复牙胶电子休息。可以肯定，这是滴水不漏。等到胶水是用钳子测试它干又硬。 注:另外一个灌溉加速固化过程。 </li><li>取出隆起缝线和关闭与皮肤缝线伤口。 </li><li>拉起鼠标的侧翼的皮肤上。插入皮下针，轻轻地替代0.5毫升无菌生理盐水皮下的体液平衡。 </li><li>手术后，保持动物的热回收笼90分钟。等到动物是让他们无人之前完全清醒。等待，直到动物它们返回到与其他动物的笼子前完全恢复。 </li><li>重复皮下盐水体积替代约12小时后，如在步骤1.16中描述。 </li><li>通过管饲法或直接手术（ 例如，对乙酰氨基酚（10毫克/毫升）或其他非甾体抗炎药）后总是适用镇痛到口腔。 </li><li>检查<html>动物手术后，每天的状况，始终在培养皿使饮食简单，避免了手术后的关键减肥提供泥地板上动物性食物。 注:活体显微镜可以在颅窗口制备后的第一天进行。 </li><li>应用异氟醚麻醉并固定在头支架的动物。打开皮肤缝合，并用干净棉球和无菌生理盐水的窗口。 24小时后，颅窗口应该由该时间点，这使得成像充满脑脊液。通过建立显微镜协议执行18成像。 </li></ol> 2.远端缺血注:缺血过程应CW准备后进行大约5天。这最小化从所引起的CW制备与中风引起的免疫反应的免疫反应的干扰。 "SRC ="/文件/ ftp_upload / 54701 / 54701fig1.jpg"/&gt; 图1.概述远端缺血的。答:这是关于运前的船只很好的概述。后第一次接触双极B的船只。之后第二双极接触C.的船只。 D.有关船只，目前已完全关闭的概述。 E.最后概述与较低的放大倍率。 <a href="http://ecsource.jove.com/files/ftp_upload/54701/54701fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <ol><li>麻醉使用麻醉面罩和适当麻醉政权的老鼠，在咨询的兽医人员（ 例如，诱导1.5 - 通过 1.5％异氟醚2/3 N 2 O和1/3 O 2 - 2％异氟醚和1.0保养蒸发器）。 </li><li>刮胡子，除去毛和消毒的皮肤和周围的皮毛与相应的代理（例如，70％乙醇），并随后干燥。 </li><li>保持体温的小鼠在36.5℃，在手术过程中±0.5°C。反馈控制的加热垫，根据鼠标的直肠温度下加热，强烈建议。 </li><li>放置动物中的横向位置。固定在麻醉面罩的鼻子和调整异氟烷浓度为1.0 - 1.5％。 </li><li>应用湿药膏（泛醇），以两只眼睛。 </li><li>用于CW准备做皮肤切口。 </li><li>轻轻分开皮肤与下方识别颞肌。 </li><li>调整高频发生器的能量（5 - 7 W）和使用双极性模式。 </li><li>使用电镊子和小心从头骨移除颞肌，创造了肌瓣，无需完全除去肌肉。 </li><li>确定马华在颞区的喙部分透明的头骨下方，背侧复古眼眶升窦。如果马华分叉不能被识别，试图找出最船只喙。 </li><li>薄MCA分支与microdrill同时不断冲洗，以避免热损伤上面的头骨。 </li><li>提起与插管骨​​和microforceps删除它。 </li><li>减少高频发生器的能量到3 - 5 W. </li><li>从上面的方法动脉，轻轻两侧双极钳触碰它无需提起容器。 </li><li>近端和远端凝结动脉至脉管分叉。 </li><li>等待30秒，然后轻轻触摸动脉，以确保血液流动被永久中断。如果观察到一个再通重复电。 </li><li>固定用1或2个针颞肌的肌肉插入覆盖骨缺损，如果可能的话。 </li><li>缝合伤口和动物放入加热的回收箱。一般情况下，挥发性ANEST后迅速恢复的动物hesia。 </li><li>对于体积替代，应用0.5毫升无菌盐水皮下的，如在步骤1.16中描述。 </li><li>手术后，让动物们留在加热回收笼90分钟。等到动物是让他们无人之前自觉。其中只有返回与其他动物的笼子时，他们完全康复。 </li><li>重复体积的取代，如在步骤1.16所解释的，经过12小时。 </li><li>申请通过饮水（ 例如，对乙酰氨基酚（10毫克/毫升）或其他非甾体抗炎药）术后镇痛。 </li><li>每天检查手术后的动物的医疗条件。在地板上的培养皿提供捣碎动物食品，以简化的饮食，并尽量减少术后体重减轻。 </li></ol> 3.假治疗<ol><li>相同执行所有程序，步骤1和2，上面包括CW准备 - 除了不凝结暴露大脑中描述tery。 </li></ol>

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The authors have nothing to disclose.

中风是长期的伤残和死亡全世界1的主要原因之一。除了急性期治疗，研究新的方法和机制，加快和提高中风后恢复仍然是一个主要的医疗目标7。试验行程研究者经常使用局灶性脑缺血啮齿动物模型。实际上，模型诱导瞬时或永久MCAO模仿最常见的类型患者4局灶性脑缺血中的一个。除了MCA的近端闭塞，为远侧MCAO手术闭塞灯丝模型可能是在实验性卒中研究5,19中最常?...

Stroke is among the principal causes of long-term disability and death worldwide1, coming second after coronary heart disease. In addition, stroke is the primary cause of long-term disability, underscoring its tremendous socioeconomic impact6-8. Beyond acute treatment, investigating new approaches and mechanisms to accelerate and enhance recovery after stroke remains a prime medical goal7. 
In the last few decades, data from experimental stroke research has contributed substantially to understanding the complex pathophysiological cascades triggered by ischemia9,10. Excitotoxicity, apoptosis, peri-infarct depolarization, and inflammation have been identified as the most relevant mediators of cell death following focal cerebral ischemia. Moreover, using animal models of cerebral ischemia, important concepts, diagnostic modalities, and therapeutic approaches have been developed and validated (e.g., "penumbra" and thrombolysis)11.
The availability of experimental stroke models, combined with non-invasive imaging modalities (e.g., magnetic resonance imaging (MRI), computed tomography, or laser speckle contrast analysis), enables the researcher to investigate hyperacute and chronic pathophysiological changes induced by the ischemic insult in a longitudinal manner12. Along with studying the spatiotemporal profile of the evolving lesion, changes resembling neuronal plasticity can be investigated and correlated to functional outcomes and histological findings. Within the last few years, further methodological advances have been made using the combination of cerebral ischemia models and in vivo microscopy via cranial windows13. These new techniques allow investigators to analyze the neurovascular unit at the cellular and molecular level, with great analytic power in the acute, subacute, and chronic phases following focal cerebral ischemia14. Moreover, in vivo microscopy imaging of microcirculatory dynamics has revealed novel aspects of cerebral microvasculature function and angioarchitecture, with significant pathophysiological relevance15-17. 
In this protocol, we present how to perform a chronic CW preparation lateral to the sagittal sinus and how to surgically induce a distal stroke underneath the window. This mouse model can be applied to sequential imaging of acute, subacute, and chronic changes following focal cerebral ischemia via epi-illuminating, confocal laser scanning, and two-photon intravital microscopy.

<table><tbody><tr><td>Binocular surgical microscope</td><td>Zeiss</td><td>Stemi 2000 C</td><td></td></tr><tr><td>Light source for microscope</td><td>Zeiss</td><td>CL 6000 LED</td><td></td></tr><tr><td>Heating pad with rectal probe</td><td>FST</td><td>21061-10</td><td></td></tr><tr><td>Stereotactic frame</td><td>Kopf</td><td>Model 930</td><td></td></tr><tr><td>Anaethesia system for isoflurane</td><td>Draeger</td><td></td><td></td></tr><tr><td>Isoflurane</td><td>Abott</td><td></td><td></td></tr><tr><td>Dumont forceps #5</td><td>FST</td><td>11251-10</td><td></td></tr><tr><td>Dumont forceps #7</td><td>FST</td><td>11271-30</td><td></td></tr><tr><td>Bipolar Forceps</td><td>Erbe</td><td>20195-501</td><td></td></tr><tr><td>Bipolar Forceps&amp;nbsp;</td><td>Erbe&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; 20195-022</td><td></td><td></td></tr><tr><td>Microdrill</td><td>FST&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; 18000-17&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td></td><td></td></tr><tr><td>Needle holder</td><td>FST</td><td>12010-14</td><td></td></tr><tr><td>5-0 silk suture</td><td>Feuerstein, Suprama</td><td></td><td></td></tr><tr><td>7-0 silk suture</td><td>Feuerstein,Suprama</td><td></td><td></td></tr><tr><td>8-0 silk suture</td><td>Feuerstein, Suprama</td><td></td><td></td></tr><tr><td>Veterinary Recovery Chamber</td><td>Peco Services</td><td>V1200</td><td></td></tr></tbody></table>

lateral chronic cranial window preparation enables in vivo observation following distal middle cerebral artery occlusion in mice

局灶性脑缺血（即缺血性中风）可能会导致重大的脑损伤，导致神经功能严重受损并因此导致运动和认知障碍的主机。它的高患病率带来了严重的健康负担，中风是长期的伤残和死亡全世界1的主要原因之一。神经功能的恢复是，在大多数情况下，只有部分。迄今为止，治疗选择是非常有限的，尤其是由于对溶栓2,3窄时间窗。确定的方法来加速从中风复苏仍尖端医疗的目标;然而，这已经阻碍了机械见解不足到恢复过程。试验行程研究者经常使用局灶性脑缺血啮齿动物模型。除了急性期，中风的研究越来越集中于亚急性期和慢性期脑缺血。大多数研究者中风申请永久或TRAN在小鼠或大鼠大脑中动脉闭塞过性。在患者中，MCA的闭塞是缺血性中风4的最常见的原因之一。除了使用灯丝模型MCA的近端闭塞，远端MCA的外科闭塞可能是在实验性卒中研究5中最常用的模型。远端（到lenticulo-纹动脉的分支）马华支闭塞通常免去了纹状体和主要影响大脑皮层。血管闭塞可以是永久的或暂时的。相对于长期预后病变体积和非常低的死亡率高重复性是这种模式的主要优点。在这里，我们将演示如何进行慢性颅窗口（CW）准备外侧为矢状窦，事后如何手术诱导采用开颅手术方法的窗口下方的远端行程。这种方法可应用于以下通过缺血的急性和慢性的变化序列成像外延照射，共焦激光扫描，和双光子活体显微镜。

时间轴和有代表性的结果示于图2和3。颅窗口制备，用小颅窗口外侧上矢状窦（ 图2 B，C，D）的结果在由有经验的外科医生进行的时非常低的死亡率和发病率。所有10只动物存活，所有慢性的CW可用于高质量成像，甚至手术后28天。有一个与伤口感染或其他并发症没有问题。 由于短曝光挥发...

Watch this Scientific Journal Video about Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice at JoVE.com

Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice

远端大脑中动脉分支（缺血）的手术闭塞是实验性卒中研究常用的模式。该原稿描述永久MCAO的基本技术，有横向颅窗口的插入，这提供了在小鼠中纵向活体显微镜的机会相结合。

横向慢性颅窗口准备启用<em&gt;在体内</em&gt;观察继末节小鼠大脑中动脉闭塞

Focal cerebral ischemia (i.e., ischemic stroke) may cause major brain injury, leading to a severe loss of neuronal function and consequently to a host of ...

lateral-chronic-cranial-window-preparation-enables-vivo-observation

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice

11.6K 次观看.  Charité-Universitätsmedizin. 远端大脑中动脉分支（缺血）的手术闭塞是实验性卒中研究常用的模式。该原稿描述永久MCAO的基本技术，有横向颅窗口的插入，这提供了在小鼠中纵向活体显微镜的机会相结合。 

视频: Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice

Intravital Microscopy of the Mouse Brain Microcirculation using a Closed Cranial Window

This experimental model was designed to assess the mouse pial microcirculation during acute and chronic, physiological and pathophysiological hemodynamic, inflammatory and metabolic conditions, using in vivo fluorescence microscopy. A closed cranial window is placed over the left parieto-occipital cortex of the mice. Local microcirculation is recorded in real time through the window using epi and fluorescence illumination, and measurements of vessels diameters and red blood cell (RBC) velocities are performed. RBC velocity is measured using real-time cross-correlation and/or fluorescent-labeled erythrocytes. Leukocyte and platelet adherence to pial vessels and assessment of perfusion and vascular leakage are made with the help of fluorescence-labeled markers such as Albumin-FITC and anti-CD45-TxR antibodies. Microcirculation can be repeatedly video-recorded over several days. We used for the first time the close window brain intravital microscopy to study the pial microcirculation to follow dynamic changes during the course of Plasmodium berghei ANKA infection in mice and show that expression of CM is associated with microcirculatory dysfunctions characterized by vasoconstriction, profound decrease in blood flow and eventually vascular collapse.

Intravital microscopy to follow temporal and spatial hemodynamic and inflammatory events in the pial microcirculation.

活体鼠脑微循环显微镜使用一个封闭的颅窗口

This experimental model was designed to assess the mouse pial microcirculation during acute and chronic, physiological and pathophysiological hemodynamic, ...

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 ...

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. ...

Utilizing a Cranial Window to Visualize the Middle Cerebral Artery During Endothelin-1 Induced Middle Cerebral Artery Occlusion

Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain1-3. This technique can be performed in many locations overlying the rat cerebrum, but is most easily carried out by creating a craniectomy over the readily accessible frontal or parietal bones. Most frequently, we have used this technique in combination with the endothelin-1 middle cerebral artery occlusion model of ischemic stroke to quantify the changes in middle cerebral artery vessel diameter that occur with injection of endothelin-1 into the brain parenchyma adjacent to the proximal MCA4, 5. In order to visualize the proximal portion of the MCA during endothelin -1 induced MCAO, we use a technique to create a cranial window through the temporal bone on the lateral aspect of the rat skull (Figure 1). Cerebral arteries can be visualized either with the dura intact or with the dura incised and retracted. Most commonly, we leave the dura intact during visualization since endothelin-1 induced MCAO involves delivery of the vasoconstricting peptide into the brain parenchyma. This bypasses the need to incise the dura directly over the visualized vessels for drug delivery. This protocol will describe how to create a cranial window to visualize cerebral arteries in a step-wise fashion, as well as how to avoid many of the potential pitfalls pertaining to this method.

This article describes a method for visualizing rat cerebral arteries through a cranial window using temporal craniectomy in order to view proximal portions of the middle cerebral artery (Figure 1). This versatile method can be combined with various techniques of drug delivery to measure cerebral artery reactivity in vivo.

利用颅窗口，在可视化的大脑中动脉内皮素-1诱导大脑中动脉闭塞

Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain1-3. This technique can be ...

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 ...

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 ...

建立经颅大脑中动脉闭塞小鼠模型

A Modified Transcranial Middle Cerebral Artery Occlusion Model to Study Stroke Outcomes in Aged Mice

In experimental stroke research, middle cerebral artery occlusion (MCAO) with an intraluminal filament is widely used to model ischemic stroke in mice. The filament MCAO model typically exhibits a massive cerebral infarction in C57Bl/6 mice that sometimes includes brain tissue in the territory supplied by the posterior cerebral artery, which is largely due to a high incidence of posterior communicating artery atresia. This phenomenon is considered a major contributor to the high mortality rate observed in C57Bl/6 mice during long-term stroke recovery after filament MCAO. Thus, many chronic stroke studies exploit distal MCAO models. However, these models usually produce infarction only in the cortex area, and consequently, the assessment of post-stroke neurologic deficits could be a challenge. This study has established a modified transcranial MCAO model in which the MCA at the trunk is partially occluded either permanently or transiently via a small cranial window. Since the occlusion location is relatively proximal to the origin of the MCA, this model generates brain damage in both the cortex and striatum. Extensive characterization of this model has demonstrated an excellent long-term survival rate, even in aged mice, as well as readily detectable neurologic deficits. Therefore, the MCAO mouse model described here represents a valuable tool for experimental stroke research.

作者聚焦:中风研究的创新技术和未来方向 

This protocol demonstrates a unique mouse stroke model with a medium-sized infarct and an excellent survival rate. This model allows preclinical stroke researchers to extend the ischemia duration, use aged mice, and assess long-term functional outcomes.

改进的经颅大脑中动脉闭塞模型研究老年小鼠脑卒中结局

In experimental stroke research, middle cerebral artery occlusion (MCAO) with an intraluminal filament is widely used to model ischemic stroke in mice. ...

横向慢性颅窗口准备启用

摘要

探索更多视频

此视频中的章节

活体鼠脑微循环显微镜使用一个封闭的颅窗口

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

小鼠模型的腔内MCAO脑梗塞评价甲酚紫染色

利用颅窗口，在可视化的大脑中动脉内皮素-1诱导大脑中动脉闭塞

对大脑中动脉闭塞再灌注损伤诱发中风小鼠的手术方法

脑缺血再灌注的双血管闭塞小鼠模型

改进的经颅大脑中动脉闭塞模型研究老年小鼠脑卒中结局

改进的经颅大脑中动脉闭塞模型研究老年小鼠脑卒中结局

改进的经颅大脑中动脉闭塞模型研究老年小鼠脑卒中结局