This work was supported by National Institutes of Health grants HL087990 (G.L.).

道德声明:成年雄性SD大鼠（体重330-380克），在此协议中使用。该协议被批准的机构动物护理和使用委员会（IACUC）在路易斯安那州立大学健康科学中心 - 什里夫波特，是符合"指南的护理和使用实验动物"（第八版，国家科学院，2011 ）。 1，制备改性PE-50管<ol><li>保持30厘米长的PE-50管上方的气体火手，渐渐软化管。轻轻拉伸管。 </li><li>选择使用数字卡尺（ 图1）拉伸管的一个点，它标记，并削减了改性PE-50管到25厘米长的段有1厘米长的尖（外直径:0.30-0.34毫米）。 </li></ol> 2，准备同源血块<ol><li>麻醉与异氟醚（感应5％，为维护2-3％）的70％，N 2 O和30％的澳鼠&lt;子&gt; 2通过在采血前面罩。确认麻醉用脚趾捏。 </li><li>与已发表 ​​的方法进行4供体大鼠股动脉插管，并直接传输股动脉的血变成20厘米长的PE-50管。将管2小时，在室温下凝块血，然后保持该管22小时，在4℃。注:插管使用无菌技术，如步骤3.1所述进行。供体鼠被回收，用于进一步取样。 </li><li>切PE-50管成50mm的段和冲洗掉从该管的凝块到含有生理盐水的无菌培养皿中。 </li><li>传送50mm长凝块成60毫米长的PE-10管和PE-10管的每一端连接到包含有23政针生理盐水20厘米长的PE-50管连接至1ml注射器（ 图2） 。 </li><li>从一个针筒转移凝块由连续交替运动到另一个5分钟，这可以从除去血细胞凝块和最小化的脆弱血管外凝结的血块，而不破坏纤维蛋白核心。 </li><li>切血块成40毫米长的段，如在无菌条件下，在步骤1.2中所述的血块段式转换成改性PE-50导管。注:所有的PE管进行灭菌与环氧乙烷。 </li></ol> 3，栓塞大脑中动脉闭塞（图3A，B） <ol><li>通过高压灭菌（最小121℃，15磅，15分钟）灭菌所有的外科手术工具。消毒手术表，并使用70％的乙醇相关的外科手术设备。 </li><li>麻醉与异氟醚对大鼠按照步骤2.1所述。测试麻醉深度上都前后脚进行脚趾捏和观察到的任何动作（收回爪子）表示该动物是没有足够的麻醉做手术。适用于双眼少量兽医药膏，以防止干燥时在麻醉下。剃了毛的腹侧颈和头重地区码与电推剪到皮肤区域暴露。 </li><li>将大鼠在一个加热垫仰卧位。插入直肠探头，控制和使用恒温电热毯控制单元之间保持36.5-37.5℃的体温。 </li><li>消毒的剃光皮肤和周围的毛，用10％聚维酮碘接着用70％乙醇。 </li><li>请在解剖显微镜下脖子上有2厘米长的正中切口。使用牵引器，以暴露手术野和解剖右颈总动脉（CCA），颈外动脉（ECA），颈内动脉（ICA），以及翼腭动脉（PPA）的分类从周围神经和筋膜（ 图3A）。注意事项:手术前，改变无菌手套，如果没有助手帮助准备的动物。 </li><li>解剖CCA不受周围神经（不伤害迷走神经），然后将动脉下无菌5-0丝线缝合。打个活结（＃1），用一个小hemos把握缝合李永达和拉教授向身体。 </li><li>剖析非洲经济委员会和它的两个分支，枕动脉（OA）和甲状腺上动脉（STA）。使用兽药电单元凝结两个分支。将两片无菌6-0丝线缝合非洲经委会下。 </li><li>分开的两个丝的动脉，朝向头部一体成型（前端），另一个朝向身下放置。领带系得过紧（＃2）在最近的一侧头部，抓住丝绸小止血和拉教授向头部。准备一个松散的结（＃3）与其他丝绸以后使用。 </li><li>解剖ICA和PPA从周围神经（而不损害迷走神经）。领带的PPA与6-0缝线（＃4）。做一个活结与周围的ICA（＃5），6-0缝合。另外，采用微血管夹子夹在ICA。 </li><li>切一小口（半直通）到非洲经委会紧（＃2）和一个Vannas式弹簧剪刀宽松（＃3）结扎线之间。将改性PE-50桶ê含血凝块放入切口并进入文建会分叉。拧松的绑带（＃3）围绕所述内腔刚够固定保持在留置管的迁移率。 </li><li>切断非洲经委会在小孔部位释放的树桩和定位ECA和ICA分叉下方的树桩;这将允许改性管容易滑入ICA。打开ICA和轻轻推进从ECA的腔管进入ICA，直到导管的前端到达MCA的原点（〜17毫米的分叉处）。注意:在插入管的末端进入非洲经委会，管，用70％乙醇消毒外表面，然后洗净，用无菌生理盐水。 </li><li>注入通过改性PE-50导管的血块随着10微升生理盐水在10秒内用100微升Hamilton注射器（ 图3B）。 </li><li>从非洲经委会5分钟后拔出导管。配合非洲经委会并重新打开CCA。缝合切口在颈部。 </li></ol> 4，监测局部脑血流量（rCBF） <ol><li>在此之前大脑中动脉闭塞，使1.2厘米长的正中切口在头皮暴露颅骨。用牙刮和无菌棉互换取下颅骨骨组织。注意:在手术前，暴露的头皮区域和周围的毛皮被消毒，用10％聚维酮碘接着用70％乙醇。 </li><li>钻设为2mm后到5毫米的横向使用0.7mm的球形不锈钢毛刺（ 图4）的前囟1.5毫米直径的钻孔。 注意:保持硬脑膜完整。 </li><li>把探头0.5毫米硬脑膜表面之上。监测脑血流量在0（基线），5，15，30，60，90，和栓塞后120分钟，并继续在5，15，30，45，和tPA的静脉内给药后的60分钟。脑血流量的最后测量之后，对头皮切口被缝合闭合。注:在颈部切开我们测量脑血流量为马华occlus前的基​​线离子。发表大脑中动脉闭塞，我们衡量颈部切口缝合关闭后局部脑血流量。因此，颈部切口保持无菌。 </li></ol> 5，术后护理<ol><li>注入2.5的生理盐水溶液皮下注射毫升，以防止脱水，并在外科手术，并根据需要缓解疼痛的每6-12小时后，立即注入BUPRENEX（0.05毫克/千克，皮下注射）。 </li><li>停止异氟醚麻醉。将鼠在37℃兽医恢复室（让动物温暖），并保持观察。它一般需要10分钟的大鼠从麻醉中恢复。然后，把动物带进无菌笼子里，放置一些湿的食物在培养皿中，在笼子里，然后返回笼子的动物消毒室。 </li><li>中风后24小时，安乐死的大鼠用戊巴比妥钠（100毫克/千克体重，腹腔注射）过量。 </li></ol> 6，神经功能缺损评分<ol><li>之前和在栓塞后2小时进行Bederson得分。使用分级Š0-3 Cale的先前所描述的5:0，由尾提高大鼠，延伸的两个前肢在地上，没有表现出其它缺陷的动物（正常运动）; 1，通过尾部提高大鼠，弯曲对侧前肢; 2，降低抗侧推（和前肢屈曲），而不盘旋; 3，同样的行为，与盘旋2级。 注:显示大鼠在栓塞后2小时Bederson评分= 0（无赤字）被排除在进一步的研究中。 </li></ol>

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We do not have any potential conflicting interests to disclose.

在这项研究中，我们发现在大鼠，在马华的起源是由纤维蛋白凝块丰富闭塞进行栓塞缺血中风模型的标准方法。这种模式的主要优点是:MCA的用富含纤维蛋白的​​血凝块杆的咬合是类似于在人类中血栓栓塞性中风，栓塞中风模型是适合于执行溶栓治疗的临床前研究，并且该模型可以开发由民政部提供的境内可重复和可预测的梗死体积。 用于进行栓塞MCAO模型，该凝块引入，稳...

A correction was made to <a href="/video/51956">Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats</a>. The institution information was updated.
The institution "Louisiana State University Health Science Center" was changed to "Louisiana State University Health Science Center, Shreveport".

A correction was made to <a href="/video/51956">Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats</a>. The institution information was updated.

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

中风是死亡，在美国的第三大原因，但对于急性中风的治疗选择仍然有限。目前，静脉内输注重组组织型纤溶酶原激活物（tPA）的溶解血块是最有效的治疗急性缺血性中风。然而，tPA的使用是由它的治疗窗狭窄和脑出血的危险性增加的限制。因此，迫切需要行程适合溶栓研究的典范。 大脑中动脉（MCA）是最常见的吸留在中风在人类的动脉。集中在这个动脉缺血性中风的许多动物模型已经建立。目前，已开发的啮齿动物脑缺血模型通过阻断马华两大类:缝合缺血模型和栓塞性脑缺血模型。虽然经由所述管腔内缝合技术MCAO模型已被广泛地应用于机械驱动冲程的研究，这些缝合模式不模仿人的行程，因为高达80％的人的中风是由血栓或栓塞所引起的。但是，使用血栓栓塞性中风模型密切模仿人类的​​行程，被认为适合溶栓研究。大鼠被首先开发这栓塞模型Overgaard 等。1于1992年，其特征在于:张某等人在1997年第2和Dinapoli 等人于2006年3虽然栓塞缺血已经获得了越来越多的关注，也有面临技术难题许多实验室。 在本文中，我们展示了用于生产栓塞缺血与同源血块在成年大鼠，可开发马华境内可预测心肌梗死的标准方法。在这篇文章中介绍的技术将有助于研究人员克服技术难题，为建立这个模型中风的研究。

<table><tbody><tr><td>rh-tPA</td><td>Chemical</td><td>Genentech</td><td></td></tr><tr><td>2,3,5-triphenyltetrazolium chloride</td><td>Chemical</td><td>Sigma</td><td>T8877</td></tr><tr><td>Anesthesia vaporizer</td><td>Equipment</td><td>Soma Technology</td><td>Drager Vapor 19.1&amp;nbsp;&amp;nbsp;</td></tr><tr><td>Rechargeable high speed micro drill</td><td>Equipment</td><td>Fine Science Tools</td><td>18000-17</td></tr><tr><td>Curved scissors</td><td>Equipment</td><td>Fine Science Tools</td><td>14117-14</td></tr><tr><td>Dumont forceps (Medical #7)</td><td>Equipment</td><td>Fine Science Tools</td><td>11270-20</td></tr><tr><td>Dumont forceps (Medical #5)</td><td>Equipment</td><td>Fine Science Tools</td><td>11251-35</td></tr><tr><td>Vannas-style spring scissors</td><td>Equipment</td><td>Fine Science Tools</td><td>15000-03</td></tr><tr><td>Veterinary recovery chamber&amp;nbsp;</td><td>Equipment&amp;nbsp;</td><td>Peco Services&amp;nbsp;</td><td>V1200&amp;nbsp;&amp;nbsp;</td></tr><tr><td>Genie plus syringe pump</td><td>Equipment</td><td>Kent Scientific Corporation</td><td></td></tr><tr><td>Rat brain matrix</td><td>Equipment</td><td>Kent Scientific Corporation</td><td>RBMA-310C</td></tr><tr><td>Digital caliper&amp;nbsp;</td><td>Equipment</td><td>World Precision Instruments</td><td>501601</td></tr><tr><td>Dissecting microscope</td><td>Equipment</td><td>World Precision Instruments</td><td>PZMTIII-BS-LWD</td></tr><tr><td>Hamilton syringe</td><td>Equipment</td><td>Hamilton</td><td>model 710</td></tr><tr><td>Homeothermic blanket control unit</td><td>Equipment</td><td>Harvard Apparatus</td><td></td></tr><tr><td>Electric clipper</td><td>Equipment</td><td>Braintree Scientific</td><td>CLP-9931</td></tr><tr><td>Veterinary electrosurgical unit</td><td>Equipment</td><td>MACAN Manufacturing Company</td><td>MV-9</td></tr><tr><td>Blood flowmeter</td><td>Equipment</td><td>Adinstruments</td><td></td></tr><tr><td>PowerLab 4/30</td><td>Equipment</td><td>Adinstruments</td><td></td></tr><tr><td>LabChart 7.2</td><td>software</td><td>Adinstruments</td><td></td></tr><tr><td>1 ml syringe</td><td>Consumable</td><td>Becton, Dickinson and Company</td><td>309659</td></tr><tr><td>23 G needle</td><td>Consumable</td><td>Becton, Dickinson and Company</td><td>305143</td></tr><tr><td>30 G needle</td><td>Consumable</td><td>Becton, Dickinson and Company</td><td>305106</td></tr><tr><td>PE-50 tubing</td><td>Consumable</td><td>Becton, Dickinson and Company</td><td>427517</td></tr><tr><td>PE-10 tubing</td><td>Consumable</td><td>Becton, Dickinson and Company</td><td>427400</td></tr><tr><td>6-0 Silk suture</td><td>Consumable</td><td>Harvard apparatus</td><td>723287</td></tr><tr><td>5-0 Silk suture</td><td>Consumable</td><td>Harvard Apparatus</td><td>517607</td></tr></tbody></table>

embolic middle cerebral artery occlusion (mcao) for ischemic stroke with homologous blood clots in rats

在临床上，溶栓治疗用重组组织型纤溶酶原激活剂（tPA）的仍然是最有效的治疗急性缺血性中风。然而，tPA的使用是由它的治疗窗狭窄和出血性转化的危险性增加的限制。目前迫切需要开发适合中风模型来研究新的溶栓药物和战略进行治疗缺血性中风的。目前，缺血性脑卒中模型两大类，已经开发了大鼠和小鼠:线栓缺血及栓塞缺血。虽然经由所述管腔内缝合技术MCAO模型已被广泛地应用于机械驱动冲程的研究，这些缝合模型不模仿临床情况，因此并不适合用于血栓溶解研究。在这些模型中，栓塞MCAO模型密切模仿人的缺血性中风和适合溶栓治疗的临床前研究。大鼠OV最早开发此栓塞模型ergaard 等。1于1992年，其特征在于:张某等人于1997年2虽然栓塞缺血已经获得了越来越多的关注，也有面临许多实验室的技术问题。为了满足溶栓的研究日益增加的需求，我们提出栓塞缺血的高度重复性的模型大鼠，可开发马华境内可预见的梗死体积。简言之，改性PE-50管轻轻地从颈外动脉（ECA）被推进到颈内动脉（ICA），直至导管尖端的内腔到达MCA的来源。通过导管，单个同源血液凝块被放置在MCA的来源。为了找出大脑中动脉闭塞的成功，局部脑血流监测，神经功能缺损及梗死体积进行了测量。本文提出的技术将有助于研究人员克服了建立这一模式中风研究技术问题。&lt;/ P&gt;

激光多普勒血流仪（LDF）用于诱导脑缺血6,7的过程中监测脑血流量。许多实验室，包括我们的实验室一直在使用局部脑血流量来识别动物的成功大脑中动脉闭塞，但基线的阈值，这是关系到测量现场实验室之间变化。探针的LDF被定位为2mm后方和5mm横向于前囟如前面6所述。脑血流量在0监测，5，15，30，60，90，和120分钟注射凝块后。在这个数据的基础上，只有表现出局部脑血流量减?...

Watch this Scientific Journal Video about Embolic Middle Cerebral Artery Occlusion MCAO for Ischemic Stroke with Homologous Blood Clots in Rats at JoVE.com

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 ...

embolic-middle-cerebral-artery-occlusion-mcao-for-ischemic-stroke

Research

JoVE Journal

Medicine

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

27.0K 次观看.  Louisiana State University Health Science Center, Shreveport. 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. 

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

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

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

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 20091, 2. Several models of cerebral ischemia have been developed to mimic the human condition of stroke. It has been suggested that up to 80% of all strokes result from ischemic damage in the middle cerebral artery (MCA) area3. In the early 1990s, endothelin-1 (ET-1) 4 was used to induce ischemia by applying it directly adjacent to the surface of the MCA after craniotomy. Later, this model was modified 5 by using a stereotaxic injection of ET-1 adjacent to the MCA to produce focal cerebral ischemia. The main advantages of this model include the ability to perform the procedure quickly, the ability to control artery constriction by altering the dose of ET-1 delivered, no need to manipulate the extracranial vessels supplying blood to the brain as well as gradual reperfusion rates that more closely mimics the reperfusion in humans5-7. On the other hand, the ET-1 model has disadvantages that include the need for a craniotomy, as well as higher variability in stroke volume8. This variability can be reduced with the use of laser Doppler flowmetry (LDF) to verify cerebral ischemia during ET-1 infusion. Factors that affect stroke variability include precision of infusion and the batch of the ET-1 used6. Another important consideration is that although reperfusion is a common occurrence in human stroke, the duration of occlusion for ET-1 induced MCAO may not closely mimic that of human stroke where many patients have partial reperfusion over a period of hours to days following occlusion9, 10. This protocol will describe in detail the ET-1 induced MCAO model for ischemic stroke in rats. It will also draw attention to special considerations and potential drawbacks throughout the procedure.

Several animal models of cerebral ischemia have been developed to simulate the human condition of stroke. This protocol describes the endothelin-1 (ET-1) induced middle cerebral artery occlusion (MCAO) model for ischemic stroke in rats. In addition, important considerations, advantages, and shortcomings of this model are discussed.

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 ...

2-Vessel Occlusion/Hypotension: A Rat Model of Global Brain Ischemia

Cardiac arrest followed by resuscitation often results in dramatic brain damage caused by ischemia and subsequent reperfusion of the brain. Global brain ischemia produces damage to specific brain regions shown to be highly sensitive to ischemia 1. Hippocampal neurons have higher sensitivity to ischemic insults compared to other cell populations, and specifically, the CA1 region of the hippocampus is particularly vulnerable to ischemia/reperfusion 2.
The design of therapeutic interventions, or study of mechanisms involved in cerebral damage, requires a model that produces damage similar to the clinical condition and in a reproducible manner. Bilateral carotid vessel occlusion with hypotension (2VOH) is a model that produces reversible forebrain ischemia, emulating the cerebral events that can occur during cardiac arrest and resuscitation. We describe a model modified from Smith et al. (1984) 2, as first presented in its current form in Sanderson, et al. (2008) 3, which produces reproducible injury to selectively vulnerable brain regions 3-6. The reliability of this model is dictated by precise control of systemic blood pressure during applied hypotension, the duration of ischemia, close temperature control, a specific anesthesia regimen, and diligent post-operative care. An 8-minute ischemic insult produces cell death of CA1 hippocampal neurons that progresses over the course of 6 to 24 hr of reperfusion, while less vulnerable brain regions are spared. This progressive cell death is easily quantified after 7-14 days of reperfusion, as a near complete loss of CA1 neurons is evident at this time.
In addition to this brain injury model, we present a method for CA1 damage quantification using a simple, yet thorough, methodology. Importantly, quantification can be accomplished using a simple camera-mounted microscope, and a free ImageJ (NIH) software plugin, obviating the need for cost-prohibitive stereology software programs and a motorized microscopic stage for damage assessment.

Bilateral carotid occlusion coupled with systemic hypotension produces global brain ischemia in the rat, resulting in damage to the hippocampus with reproducible severity. Animal subjects are impaired with predictable patterns of brain damage, they recover expediently, and mortality rates are comparatively low.

2血管闭塞/低血压：全脑缺血模型大鼠

Cardiac arrest followed by resuscitation often results in dramatic brain damage caused by ischemia and subsequent reperfusion of the brain. Global brain ...

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 ...

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow (ischemia) 1. Middle cerebral artery (MCA) delivers a great majority of blood to the lateral surface of the cortex 2, is the most common site of human stroke 3, and ischemia within its territory can result in extensive dysfunction or death 1,4,5. Survivors of ischemic stroke often suffer loss or disruption of motor capabilities, sensory deficits, and infarct. In an effort to capture these key characteristics of stroke, and thereby develop effective treatment, a great deal of emphasis is placed upon animal models of ischemia in MCA.
Here we present a method of permanently occluding a cortical surface blood vessel. We will present this method using an example of a relevant vessel occlusion that models the most common type, location, and outcome of human stroke, permanent middle cerebral artery occlusion (pMCAO). In this model, we surgically expose MCA in the adult rat and subsequently occlude via double ligature and transection of the vessel. This pMCAO blocks the proximal cortical branch of MCA, causing ischemia in all of MCA cortical territory, a large portion of the cortex. This method of occlusion can also be used to occlude more distal portions of cortical vessels in order to achieve more focal ischemia targeting a smaller region of cortex. The primary disadvantages of pMCAO are that the surgical procedure is somewhat invasive as a small craniotomy is required to access MCA, though this results in minimal tissue damage. The primary advantages of this model, however, are: the site of occlusion is well defined, the degree of blood flow reduction is consistent, functional and neurological impairment occurs rapidly, infarct size is consistent, and the high rate of survival allows for long-term chronic assessment.

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

We describe a highly reproducible method for the permanent occlusion of a rodent major cerebral blood vessel. This technique can be accomplished with very little peripheral damage, minimal blood loss, a high rate of long-term survival, and consistent infarct volume commensurate with the human clinical population.

永久性脑血管闭塞<em&gt;通过</em&gt;双人结扎横断

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow ...

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, ...

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 ...

Middle Cerebral Artery Occlusion in Rats with Preserved Vascular Anatomy

A Rat Model of Middle Cerebral Artery Occlusion/Reperfusion Without Damaging the Anatomical Structure of Cerebral Vessels

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.

The current protocol describes a rat model of middle cerebral artery occlusion/reperfusion that preserves the anatomical structure of cerebral vessels without causing damage.

Ischemic stroke stands as the primary cause of long-term disability and mortality among adults worldwide. Animal models of ischemic stroke have ...

<meta charset="utf8"></head><body>免疫荧光染色研究大鼠 MCAO 卒中模型中的细胞死亡

Evaluating Cell Death Signaling by Immunofluorescence in a Rat Model of Ischemic Stroke

Stroke is a leading cause of death and disability worldwide. Most cases of stroke are ischemic and result from the occlusion of the middle cerebral artery (MCA). Current pharmacological approaches for the treatment of ischemic stroke are limited; therefore, novel therapies providing effective neuroprotection against ischemic injury following stroke are urgently needed. In the brain tissue following ischemic stroke, the area of the ischemic penumbra is salvageable but at risk of progressing to irreversible damage. The penumbral area surrounding the infarcted core is a conceptual target for neuroprotection. Since the blood flow is partially maintained in the penumbral area, neuronal and non-neuronal cells in this area transiently survive after stroke, and these cells that are still viable may be rescued by appropriate medical interventions. Understanding the pathophysiology of the penumbra is important in the development of neuroprotective therapies because the cell death pathways activated in the ischemic penumbra may indicate therapeutic targets, such as RUNX1 and cathepsins. These protein targets functioning as mediators of programmed cell death can be further exploited in translational research. With moderate size and similarity to the human brain, the in vivo rat model of MCA occlusion (MCAO) mimics the human ischemic stroke and offers an applicable tool to investigate the penumbral pathology, examine the cell death signaling, and evaluate the effects of potential targets in the context of MCAO. Here, we describe how to induce MCAO in rats and how to perform immunofluorescence staining for the detection of cell death signaling in the rat brain following MCAO.

<meta charset="utf8"></head><body>通过免疫荧光评估缺血性卒中大鼠模型中的细胞死亡信号

Rat in vivo models are indispensable tools to investigate the pathological mechanisms and therapeutic targets for ischemic stroke. This work will describe performing immunofluorescence staining in infarcted brain slices following middle cerebral artery occlusion (MCAO) in rats.

通过免疫荧光评估缺血性卒中大鼠模型中的细胞死亡信号

Stroke is a leading cause of death and disability worldwide. Most cases of stroke are ischemic and result from the occlusion of the middle cerebral artery ...