这项工作是支持由美国心脏协会大中华区东南亚联盟（09GRNT2060421），美国医学协会，并从大学的佛罗里达州的临床与转化科学研究所的资助。亚当麦加是一个NIH / NINDS，NRSA博士前研究员（F30 NS-060335）。高血压（T32 HL-083810）从佛罗里达州的多学科培训计划的大学，博士前奖学金罗伯特·Regenhardt收到支持。

机构动物管理和使用委员会（IACUC）在佛罗里达大学和批准该协议是符合"指南的照顾和使用的实验动物"（第八版，美国国家科学院，2011年）。 物料<ol><li>动物:8周龄，雄性，只SD大鼠（查尔斯河农场，威​​尔明顿，MA，USA），体重250〜300克，手术时间。 </li><li>麻醉<ol class="lalpha"><li>吸入麻醉系统（VetEquip公司，普莱森顿，CA，USA） </li><li>异氟醚麻醉剂（巴克斯特制药，迪尔菲尔德，IL，USA） </li></ol></li><li>立体定位系统（大卫KOPF工具，图洪加，CA，USA） <ol class="lalpha"><li>小动物立体定位系统</li><li>非破裂大鼠耳棒</li><li>气体麻醉大鼠头支架</li></ol></li><li>温度调节<ol class="lalpha"><li> BAT-12探针温度计（世界精密仪器公司，萨拉索塔，佛罗里达州，美国） </li><li> T / PUMP，TP600热毯（Gaymar工业公司，果树园公园，NY，USA） </li></ol></li><li>手术器械<ol class="lalpha"><li>手术刀，虹膜镊子手柄＃和11＃刀片，格雷夫斗牛犬钳钳，拉钩，螺丝刀，10μl注射器，26号斜口针（世界精密仪器公司，萨拉索塔，佛罗里达州，美国） </li><li>微电机的演习和立体的持有人，精髓立体定向喷油器（Stoelting，伍德戴尔，IL，USA） </li><li> 1.0毫米的圆形钻钻，1.0毫米的倒锥钻牙钻（Roboz手术仪器有限公司，公司，盖瑟斯堡，MD，USA） </li></ol></li><li>手术用品<ol class="lalpha"><li>安装螺丝0-80 X 3/32，2.4毫米轴的长度，21号导管4mm长的基座下方，套管假的（塑料，罗阿诺克，VA，USA） </li><li>李连杰义齿丙烯酸树脂和液体的（郎牙科制造有限公司，公司，威灵，IL，USA） </li><li> 3.0尼龙线（绿洲，Mettawa，IL，USA） </li><li>棉拭子，Puralube的眼膏（默飞世尔科技，匹兹堡，PA，USA） </li><li>电推剪（奥斯特，普罗维登斯，RI，USA） </li></ol></li><li>化学制品<ol class="lalpha"><li>内皮素-1（美国多肽，桑尼维尔，CA，USA） </li><li>洗必泰2％（Agrilabs，圣若瑟，MO，USA） </li><li>盐酸丁丙诺啡（赫士睿公司的Lake Forest，IL，USA） </li></ol></li><li>可视化设备<ol class="lalpha"><li>手术显微镜（塞勒仪器及制造，圣路易斯，MO，USA） </li><li>光纤照明器（TechniQuip公司，利弗莫尔，CA，USA） </li></ol></li><li>激光多普勒血流仪系统（的ADInstruments公司，科罗拉多斯普林斯，CO，USA） <ol class="lalpha"><li>圣andard铅笔探头</li><li>探头支架</li><li>血液流量计</li><li> PowerLab系统4/30与LabChart 7 </li></ol></li><li>梗死体积的测量<ol class="lalpha"><li>大鼠大脑基质（Zivic - 米勒实验室，公司，艾里逊公园，PA，USA） </li><li> 2,3,5 - 三苯四唑氯化物（Sigma-Aldrich公司，圣路易斯，密苏里州，美国），在PBS中稀释至0.05％ </li><li>平板式扫描仪（爱普生Perfection V30，爱普生美国公司，长滩，CA，USA） </li><li> Image J软件（ImageJ的1.42q软件，美国国立卫生研究院，贝塞斯达，MA，USA） </li></ol></li></ol> 1。术前步骤<ol><li>手术前，老鼠被安置在一个12:12的光/暗周期，可免费使用水和啮齿动物州城。 </li><li>麻醉诱导与4％异氟烷在100％O 2的混合气体的感应室中，直到不再大鼠提款后部爪夹送。 </li><li>一化粪池在此过程中，应保持技术，包括使用无菌手套，无菌外科手术器械，一个无菌手术铺巾11。 </li><li>电推剪剃光头顶。 </li><li>大鼠置于俯卧趴在一个温度控制的操作表面（热毯）的吸收垫的位置上。 </li><li>头被放置在立体定向装置放置气体麻醉面罩开始。 </li><li>接着，耳棒被插入并拧紧。 </li><li>在手术过程中麻醉维持在100％O 2的混合气体，用2％异氟烷。 </li><li>润滑油眼药膏的双眼，眼皮被关闭，以防止眼睛干燥，在手术过程中。 </li><li>探针插入直肠温度为37±0.5℃下，以保持恒定的动物核心温度</li><li>头牢牢的立体的移动设备，手术区洁净交替2％洗必泰和盐水洗涤三次。 </li></ol> 2。手术步骤<ol><li>中线切口用手术刀，是由覆盖在颅骨的眼睛从最尾端的方面（鼻根）之间的耳（上项线）在皮肤上。 </li><li> 3斗牛犬夹横向的皮肤，然后缩回。 </li><li>从颅骨，使多个结构可以清楚地看出，用干棉签去除结缔组织。这些措施包括，冠状缝，前囟门和右侧头骨脊。使用棉签清除血液从手术领域。 </li><li>使用手术显微镜，前囟门的位置，立体机械手进行调整，直到锁定在1.0毫米的圆形钻钻上的前囟门。 </li><li>钻牙钻，然后移动到1.6毫米前侧面，前囟门和5.2毫米。 </li><li>车针，穿透颅骨上钻洞卡恩乌拉的位置（ 图1）。不断地用棉签清除过量的碎片和血液。 </li></ol>在这一点上，一个导向套管可以被插入（步骤7）或ET-1直接注射通过牙钻通孔，可以进行（直接前进到步骤16）。 <ol start="7"><li>接着，钻的孔的3个安装螺丝钻颅骨使用1.0毫米的倒锥钻牙钻（ 图1）通过部分的厚度。在每个约1-2毫米的冠状缝和矢状缝1-2毫米的横向前额骨上钻一个孔。在顶骨钻一个孔，约2-3毫米的冠状缝和2-3毫米，侧的矢状缝同侧导管牙钻洞的后方。三0-80×3/32安装螺丝被放置在这些牙钻洞，并在套管水泥控股将提供支持。螺丝应仅是先进的2或3圈，以便不损坏硬膜。 </lI&gt; <li>导管被放置在立体定位的套管夹持器，并位于前囟。 </li><li>立体机械手调整，直到锁定在导管前囟门。套管移动到牙钻孔位于1.6毫米前壁和5.2毫米的横向于前囟。 </li><li>最后，在导向套管被降低到牙钻孔4.5毫米腹侧囟与最后的前端位置。 </li><li>激光多普勒血流探测位置（可选） <ol class="lalpha"><li>为了使用LDF监测大脑的血流量，探头支架可以放置到位置加盖指南套管用牙科水泥。 </li><li>除了小楔形"选项卡，探头支架基座的底座齐平修剪。 </li><li>探针架座，然后放置后导向套管和公正的内侧与面向的标签的内侧（ 图1）的侧颅底脊。 </li><li>探针架座和g指南套管贴在一起，用牙科水泥。 </li></ol></li><li>然后，使用牙科用粘固剂的插管固定到位。水泥是在三个螺钉与所有的接触和围绕所述套管的整个基地。 </li><li>应完全干燥的水泥之前除去的套管夹持器。这大约需要5分钟。 </li></ol>在这些步骤之后，手术切口可以关闭，可拧入导向套管和套管虚设。另外，ET-1诱导MCAO可以对大鼠进行一段时间后，从插管植入手术的恢复。对于此方法，应接着执行步骤19，并在稍后的时间，可以执行步骤14-18。要执行同样的手术过程中引导导管植入和注射ET-1，第14步进行。 <ol start="14"><li>输液注射器装入与ET-1（在PBS中稀释至80μM），然后安装在立体定向喷射器。 </li><li> "立体定位的操纵器被调整直到针尖在导向套管的边缘被归零。 </li><li>针尖降低，通过导向插管17.2毫米腹侧的导向套管的边缘的位置的。如果不使用的导向套管，针尖归零在囟和降低通过牙钻孔的位置8.7毫米腹侧囟。 </li><li> 3微升80μMET1注入的速率（每分钟）的1微升。 </li><li>注射器为3分钟，在输注后是完整的，然后慢慢地去除留在原地。 </li><li>切口6.0尼龙缝线被拧入插管和插管虚设封闭。 </li><li> （ 即丁丙诺啡在0.05-0.1毫克/公斤），应使用适当的镇痛药的剂量，手术后的恢复期间，以减少疼痛和不适。 </li><li>大鼠从手术室被移除，并放置在温暖，干燥的恢复区，以防止低温，柔软的食物和水的自由，方便地访问。 </李&gt; </ol>

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没有利益冲突的声明。

ET-1诱导脑缺血是一个既定的模式，经常使用在多个品系的实验缺血性中风。许多变量，如大鼠株，动物年龄，体温，麻醉方法和操作者的专业知识，可导致梗死体积的变异性增加时，使用此模型5中，14。然而，一些研究者已经证明，这种模式的优势，包括相对非侵入性的方法，剂量反应的脑血流ET-1，和能力，以避免完全由麻醉注射ET-1在清醒大鼠14日，15。值得注意的是，有很多...

<table border="1"><tbody><tr><td></td><td></td><td></td><td><ol><li>动物:8周龄，雄性，只SD大鼠（查尔斯河农场，威​​尔明顿，MA，USA），体重250〜300克，手术时间。 </li><li>麻醉<ol class="lalpha"><li>吸入麻醉系统（VetEquip公司，普莱森顿，CA，USA） </li><li>异氟醚麻醉剂（巴克斯特制药，迪尔菲尔德，IL，USA） </li></ol></li><li>立体定位系统（大卫KOPF工具，图洪加，CA，USA） <ol class="lalpha"><li>小动物立体定位系统</li><li>非破裂大鼠耳棒</li><li>气体麻醉大鼠头支架</li></ol></li><li>温度调节<ol class="lalpha"><li> BAT-12探针温度计（世界精密仪器公司，萨拉索塔，佛罗里达州，美国） </li><li> T / PUMP，TP600热毯（Gaymar工业公司，果树园公园，NY，USA） </li></ol></li><li>手术器械<ol class="lalpha"><li>手术刀，虹膜镊子手柄＃和11＃刀片，格雷夫斗牛犬钳钳，拉钩，螺丝刀，10μl注射器，26号斜口针（世界精密仪器公司，萨拉索塔，佛罗里达州，美国） </li><li>微电机的演习和立体的持有人，精髓立体定向喷油器（Stoelting，伍德戴尔，IL，USA） </li><li> 1.0毫米的圆形钻钻，1.0毫米的倒锥钻牙钻（Roboz手术仪器有限公司，公司，盖瑟斯堡，MD，USA） </li></ol></li><li>手术用品<ol class="lalpha"><li>安装螺丝0-80 X 3/32，2.4毫米轴的长度，21号导管4mm长的基座下方，套管假的（塑料件，罗阿诺克，VA，USA） </li><li>李连杰义齿丙烯酸树脂和液体的（郎牙科制造有限公司，公司，威灵，IL，USA） </li><li> 3.0尼龙线（绿洲，Mettawa，IL，USA） </li><li>棉签，Puralube的眼膏（费舍尔科学性C，匹兹堡，PA，USA） </li><li>电推剪（奥斯特，普罗维登斯，RI，USA） </li></ol></li><li>化学制品<ol class="lalpha"><li>内皮素-1（美国多肽，桑尼维尔，CA，USA） </li><li>洗必泰2％（Agrilabs，圣若瑟，MO，USA） </li><li>盐酸丁丙诺啡（赫士睿公司的Lake Forest，IL，USA） </li></ol></li><li>可视化设备<ol class="lalpha"><li>手术显微镜（塞勒仪器及制造，圣路易斯，MO，USA） </li><li>光纤照明器（TechniQuip公司，利弗莫尔，CA，USA） </li></ol></li><li>激光多普勒血流仪系统（的ADInstruments公司，科罗拉多斯普林斯，CO，USA） <ol class="lalpha"><li>标准铅笔探头</li><li>探头支架</li><li>血液流量计</li><li> PowerLab系统4/30与LabChart 7 </li></ol></li><li>梗死体积的测量<ol class="lalpha"><li>大鼠脑矩阵（Zivic - 米勒实验室，公司，艾里逊公园，PA，USA） </li><li> 2,3,5 - 三苯四唑氯化物（Sigma-Aldrich公司，圣路易斯，密苏里州，美国），在PBS中稀释至0.05％ </li><li>平板式扫描仪（爱普生Perfection V30，爱普生美国公司，长滩，CA，USA） </li><li> Image J软件（ImageJ的1.42q软件，美国国立卫生研究院，贝塞斯达，MA，USA） </li></ol></li></ol></td></tr></tbody></table>

endothelin-1 induced middle cerebral artery occlusion model for ischemic stroke with laser doppler flowmetry guidance in rat

2009年1，2，耗资约70亿美元在美国，中风是导致残疾和死亡的第三大原因，在世界上的头号。已经开发了几种型号的脑缺血，中风模仿人类的​​条件。曾有人建议，高达80％的所有笔划的结果由缺血性脑损伤中的大脑中动脉（MCA）的第3区。在20世纪90年代早期，内皮素-1（ET-1）4，通过将它直接相邻的表面后，在MCA开颅用于诱导缺血。后来，这个模型进行了改进，采用立体定向注射ET-1的相邻MCA局灶性脑缺血。这种模式的主要优点包括能够迅速执行过程，控制动脉收缩，通过改变剂量的ET-1交付的能力，没有需要处理的颅外血管供应血液输送到大脑，并逐渐reperfusi的利率，更接近人类再灌注损伤的5-7。另一方面，ET-1的模型具有的缺点，其中包括需要开颅，以及更高的变异性的每搏输出量8。这种变化可以减少使用激光多普勒血流仪（LDF），以验证脑缺血时ET-1次静脉滴注。变异性中风的因素影响包括输液和一批ET-1的精度，使用了6个 。另一个重要因素是，虽然再灌注损伤是一种常见的发生在人类中风，阻断ET-1诱导脑缺血的持续时间可能不会紧密地模仿人类中风病人多有局部灌注在一个小时内闭塞9天， 10。此协议中详细描述的ET-1诱导的脑缺血模型大鼠缺血性中风的。在整个过程中，它也将提请注意的特殊考虑和潜在的缺点。

1。术后神经学评估动物苏醒后，可用于广泛的测试，以评估神经功能障碍，包括平衡，握力​​，爪子配售，姿势不对称和楼梯攀登。葵花籽任务的运动和感觉功能，具有显着的相关性梗死第7卷，12是一个总的评价。在这个任务中，老鼠是定时开放和消费5葵花籽。五个种子放置在一个角落里，一个空的，干燥，塑料保持架和操纵的种子所花费的时间被记录。?...

Watch this Scientific Journal Video about Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat at JoVE.com

Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat

已经开发了几种脑缺血的动物模型来模拟人的条件中风。该协议描述了内皮素-1（ET-1）诱导大鼠大脑中动脉闭塞（MCAO）模型大鼠缺血性中风。此外，重要的考虑因素，此模型的优点和缺点，进行了讨论。

内皮素-1诱导大脑中动脉闭塞模型缺血性中风激光多普勒血流仪在大鼠指导

Stroke is the number one cause of disability and third leading cause of death in the world, costing an estimated $70 billion in the United States in ...

endothelin-1-induced-middle-cerebral-artery-occlusion-model-for

Research

JoVE Journal

Medicine

19.9K 次观看.  University of Florida. 已经开发了几种脑缺血的动物模型来模拟人的条件中风。该协议描述了内皮素-1（ET-1）诱导大鼠大脑中动脉闭塞（MCAO）模型大鼠缺血性中风。此外，重要的考虑因素，此模型的优点和缺点，进行了讨论。

视频: Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat

Bilateral Common Carotid Artery Occlusion as an Adequate Preconditioning Stimulus to Induce Early Ischemic Tolerance to Focal Cerebral Ischemia

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We have established bilateral common carotid artery occlusion as a preconditioning stimulus to induce early ischemic tolerance to transient focal cerebral ischemia in C57Bl6/J mice. In this video, we will demonstrate the methodology used for this study.

There is accumulating evidence, that ischemic preconditioning (PC) &#x2013; a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We established bilateral common carotid artery occlusion (BCCAO) as a preconditioning stimulus to induce early ischemic tolerance (IT) to transient focal cerebral ischemia (induced by middle cerebral artery occlusion, MCAO) in C57Bl6/J mice.

双侧颈总动脉闭塞足够的预处理刺激诱导局灶性脑缺血早期缺血耐受

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a ...

科研

医学

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 ...

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Cerebral perfusion monitoring has been demonstrated to improve accuracy in ischemic stroke models. Technical difficulties often limit the use of this essential tool for cerebrovascular research. In this video, an optimized system is shown to obtain a single or multi-site hemodynamic monitoring during intraluminal middle cerebral artery occlusion in rats.

优化系统的脑灌注监测在中风模型大鼠的大脑中动脉闭塞腔内

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal ...

Modeling Stroke in Mice: Permanent Coagulation of the Distal Middle Cerebral Artery

Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical options are available for a small proportion of stroke patients in the acute phase. Current research is intensively searching for novel therapeutic strategies and is increasingly focusing on the sub-acute and chronic phase after stroke because more patients might be eligible for therapeutic interventions in a prolonged time window. These delayed mechanisms include important pathophysiological pathways such as post-stroke inflammation, angiogenesis, neuronal plasticity and regeneration. In order to analyze these mechanisms and to subsequently evaluate novel drug targets, experimental stroke models with clinical relevance, low mortality and high reproducibility are sought after. Moreover, mice are the smallest mammals in which a focal stroke lesion can be induced and for which a broad spectrum of transgenic models are available. Therefore, we describe here the mouse model of transcranial, permanent coagulation of the middle cerebral artery via electrocoagulation distal of the lenticulostriatal arteries, the so-called &#8220;coagulation model&#8221;. The resulting infarct in this model is located mainly in the cortex; the relative infarct volume in relation to brain size corresponds to the majority of human strokes. Moreover, the model fulfills the above-mentioned criteria of reproducibility and low mortality. In this video we demonstrate the surgical methods of stroke induction in the &#8220;coagulation model&#8221; and report histological and functional analysis tools.

Various murine models of middle cerebral artery occlusion (MCAO) are widely used in experimental brain research. Here, we demonstrate the model of transcranial permanent distal MCAO which produces consistent cortical infarction of a size corresponding to damage imposed by the majority of human ischemic strokes.

造型中风小鼠:在远端大脑中动脉永久性混凝

Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical ...

Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic stroke. However, the use of tPA is limited by its narrow therapeutic window and by increased risk of hemorrhagic transformation. There is an urgent need to develop suitable stroke models to study new thrombolytic agents and strategies for treatment of ischemic stroke. At present, two major types of ischemic stroke models have been developed in rats and mice: intraluminal suture MCAO and embolic MCAO. Although MCAO models via the intraluminal suture technique have been widely used in mechanism-driven stroke research, these suture models do not mimic the clinical situation and are not suitable for thrombolytic studies. Among these models, the embolic MCAO model closely mimics human ischemic stroke and is suitable for preclinical investigation of thrombolytic therapy. This embolic model was first developed in rats by Overgaard et al.1 in 1992 and further characterized by Zhang et al. in 19972. Although embolic MCAO has gained increasing attention, there are technical problems faced by many laboratories. To meet increasing needs for thrombolytic research, we present a highly reproducible model of embolic MCAO in the rat, which can develop a predictable infarct volume within the MCA territory. In brief, a modified PE-50 tube is gently advanced from the external carotid artery (ECA) into the lumen of the internal carotid artery (ICA) until the tip of the catheter reaches the origin of the MCA. Through the catheter, a single homologous blood clot is placed at the origin of the MCA. To identify the success of MCA occlusion, regional cerebral blood flow was monitored, neurological deficits and infarct volumes were measured. The techniques presented in this paper should help investigators to overcome technical problems for establishing this model for stroke research.

Embolic Middle Cerebral Artery Occlusion MCAO for Ischemic Stroke with Homologous Blood Clots in Rats

In this paper, we demonstrate a standard method for producing an embolic middle cerebral artery occlusion with homologous blood clots (fibrin-rich) in adult rat. This model closely mimics human ischemic stroke and is suitable for preclinical study of thrombolytic therapy for ischemic stroke.

栓塞性脑梗塞（缺血）的大鼠缺血性中风与血同源凝块

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic ...

Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week protocol for repetitive hypoxic preconditioning (RHP) induces long-term protection against central nervous system (CNS) injury in a mouse model of focal ischemic stroke. RHP consists of 9 stochastic exposures to hypoxia that vary in both duration (2 or 4 hr) and intensity (8% and 11% O2). RHP reduces infarct volumes, blood-brain barrier (BBB) disruption, and the post-stroke inflammatory response for weeks following the last exposure to hypoxia, suggesting a long-term induction of an endogenous CNS-protective phenotype. The methodology for the dual quantification of infarct volume and BBB disruption is effective in assessing neurovascular protection in mice with RHP or other putative neuroprotectants. Adult male Swiss Webster mice were preconditioned by RHP or duration-equivalent exposures to 21% O2 (i.e. room air). A 60 min transient middle cerebral artery occlusion (tMCAo) was induced 2 weeks following the last hypoxic exposure. Both the occlusion and reperfusion were confirmed by transcranial laser Doppler flowmetry. Twenty-two hr after reperfusion, Evans Blue (EB) was intravenously administered through a tail vein injection. 2 hr later, animals were sacrificed by isoflurane overdose and brain sections were stained with 2,3,5- triphenyltetrazolium chloride (TTC). Infarcts volumes were then quantified. Next, EB was extracted from the tissue over 48 hr to determine BBB disruption after tMCAo. In summary, RHP is a simple protocol that can be replicated, with minimal cost, to induce long-term endogenous neurovascular protection from stroke injury in mice, with the translational potential for other CNS-based and systemic pro-inflammatory disease states.

This protocol describes repetitive hypoxic preconditioning, or brief exposures to systemic hypoxia that reduce infarct volumes and blood-brain barrier disruption following transient middle cerebral artery occlusion in mice. It also details dual quantification of infarct volume and blood-brain barrier disruption after stroke to assess the efficacy of neurovascular protection.

神经血管保护定量继在小鼠重复缺氧预处理和瞬态大脑中动脉闭塞

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week ...

Performing Permanent Distal Middle Cerebral with Common Carotid Artery Occlusion in Aged Rats to Study Cortical Ischemia with Sustained Disability

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, obesity and diabetes. Accordingly, when evaluating therapies for stroke in animals, it is important to select a model with excellent face validity. Ischemic stroke accounts for 80% of all strokes, and the majority of these occur in the territory of the middle cerebral artery (MCA), often inducing infarcts that affect the sensorimotor cortex, causing persistent plegia or paresis on the contralateral side of the body. We demonstrate in this video a method for producing ischemic stroke in elderly rats, which causes sustained sensorimotor disability and substantial cortical infarcts. Specifically, we induce permanent distal middle cerebral artery occlusion (MCAO) in elderly female rats by using diathermy forceps to occlude a short segment of this artery. The carotid artery on the ipsilateral side to the lesion was then permanently occluded and the contralateral carotid artery was transiently occluded for 60 min. We measure the infarct size using structural T2-weighted magnetic resonance imaging (MRI) at 24 hr and 8 weeks after stroke. In this study, the mean infarct volume was 4.5% &#177; 2.0% (standard deviation) of the ipsilateral hemisphere at 24 hr (corrected for brain swelling using Gerriet&#8217;s equation, n = 5). This model is feasible and clinically relevant as it permits the induction of sustained sensorimotor deficits, which is important for the elucidation of pathophysiological mechanisms and novel treatments.

Here we present a protocol to produce permanent distal middle cerebral artery occlusion in elderly female rats with simultaneous occlusion of the carotid arteries to generate large cortical infarcts and sustained deficits. We show confirmation of the lesion size using structural MRI at 24 hr and 8 weeks after stroke.

执行永久远端大脑中与老年大鼠颈总动脉闭塞研究皮层缺血具有持续残疾

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, ...

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 ...

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.

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 ...

Transient Middle Cerebral Artery Occlusion Model of Neonatal Stroke in P10 Rats

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature and mature brain. Stroke occurs commonly in the perinatal period in humans, and transient ischemia-reperfusion is the most common form of stroke in neonates. The reperfusion phase is a critical component of injury progression, which occurs over a period of days to weeks, and of the endogenous response to injury. This postnatal day 10 (p10) rat model of transient middle cerebral artery occlusion (tMCAO) creates a unilateral, non-hemorrhagic focal ischemia-reperfusion injury that can be utilized to study the mechanisms of focal injury and repair in the full-term-equivalent brain. The injury pattern that is produced by tMCAO is consistent and highly reproducible and can be confirmed with MRI or histological analyses. The severity of injury can be manipulated through changes in occlusion time and other methods that will be discussed.

Neonatal stroke is a significant cause of early brain injury requiring a translational model with consistent focal injury patterns and high reproducibility in order to enable study. This study describes the detailed surgical procedure for creating a non-hemorrhagic, unilateral focal ischemia-reperfusion injury in full-term-equivalent rodents.

在P10大鼠新生儿中风的短暂大脑中动脉栓塞模型

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature ...

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Subtitles

内皮素-1诱导大脑中动脉闭塞模型缺血性中风激光多普勒血流仪在大鼠指导