这项工作是由欧洲共同体的第七框架计划（FP7/2007-2013）根据赠款协议N ° 201024和N ° 202213（欧洲中风网络）。 Bundesministerium献给教化和Forschung（柏林中风研究中心）和德意志研究联合会（Exzellenzcluster NEUROCURE）​​的额外资金。 作者想感谢Mareike Thielke（Dep.实验神经病学，柏林Charité）在操作过程中和埃尔克路德维希（Charité视频服务）制作动画的支持。

为了保证高质量和可重复性，我们建议使用的标准操作程序（SOP）。在这个视频，发表在我们的实验室开发和使用的SOP应用1 1。大脑中动脉闭塞<ol><li>小鼠与咨询兽医人员适当的麻醉制度麻醉。 （如诱导1.5 - 2％异氟醚和维护1.0 - 1.5％异氟醚2 / 3的N2O和1 / 3的氧气使用喷雾器）。 <ol><li>小鼠的体温保持在36.5 ° C ± 0.5 ° C在一个加热板手术。强烈建议，根据鼠标的直肠温度变暖的反馈控制的加热垫。 </li><li>一个合适的代理（如70％酒精）消毒的皮肤和周围的皮毛和事后干燥。 </li></ol></li><li>中线颈部切口及软组织拉开。 </li><li>左颈总动脉（LCCA）仔细解剖周围神经（不伤及迷走神经）和结扎是使用6.0/7.0字符串。也可用于5.0字符串。 </li><li>然后分离左侧颈外动脉（莱萨）和第二个结。 </li><li>接下来是孤立的，左侧颈内动脉（LICA）和6.0长丝准备了一个结。 </li><li>左侧颈内动脉（LICA）和左侧翼腭动脉（LPA）获得良好的查看后，两动脉被裁剪，使用微血管剪辑。 </li><li>之前，分叉的莱萨和LICA在LCCA切开一个小孔。然后引入一个单丝涂有硅固化剂混合物（见下文）的8.0尼龙LICA，直到它停在剪辑。要注意支付不进入枕动脉。 （图1） </li><li>剪切的动脉被打开，而灯丝插入到LICA闭塞Willis环的原产地的LMCA。 </li><li>第三个结对LICA封闭，固定位置的灯丝。 </li><li>小鼠接受生理盐水0.5毫升皮下注射量补货。缓解疼痛，利多卡因凝胶是在伤口局部应用。 </li><li>如果再灌注的目的是，老鼠留30 - 90分钟在激烈的笼子闭塞，可以封闭伤口，用一个小的缝合片段。之后，进行第二次麻醉，结对ICA第三是瞬间打开灯丝撤回。 </li><li>剩余的缝线缩短，适应与手术缝合皮肤。 </li><li>所有的动物都收到如上所述，第二卷补货。 </li><li>动物都放在了两个小时，体温控制在激烈的笼子。 <ol><li>动物必须每天检查，手术后不适的迹象。小鼠可能会​​表现出一定的手术减肥后。他们收到捣碎食物，在培养皿，培养皿放置在地板上，鼓励进食。七天每天食物取代。 </li></ol></li></ol> 2。假手术<ol><li>深水作业的灯丝是插入到闭塞LMCA立即取消允许即时再灌注（1.8）。随后的操作是相同的，发生脑缺血（1.9 - 1.14）的动物，包括第二次麻醉。 </li></ol> 3。灯丝的制备<ol><li>应考虑灯丝不育。事后使用消毒设备，以及适当处理灯丝是无菌手术的先决条件。消毒的灯丝是困难的，因为许多常见的灭菌方法可能会恶化长丝的质量。然而，如辐射的方法，例如紫外线或γ射线，或化学灭菌，例如高活性气体，如环氧乙烷，都适用。 </li><li> 8.0锦纶长丝被切成长度为11毫米，在显微镜下</li><li>灯丝尖端必须超过8毫米的长度，完全并均匀地涂固化剂混合物的Xantopren中号黏膜和激活的NF Optosil </li></ol> 4。代表性的成果血流量限制的期限，根据不同的电机和行为缺陷的结果。两个后30和脑缺血60分钟，在大多数情况下，动物抵抗力下降，横向推盘旋在运动障碍。温和的病变表现为一个在前面的李屈位置MBS。这些很容易观察到的迹象，可以用来作为手术成功的基本得分。 形态病变可使用或者组织学或磁共振成像（MRI）进行评估。六十分钟大脑中动脉闭塞，生产面积包括纹状体和大脑皮层组织pannecrosis，而30分钟，缺血引起主要限于纹状体的神经细胞死亡。梗死体积3（图2） ，我们预期标准偏差低于30％的一组操作。死亡率取决于阻断时间约5％后，30分钟缺血和10 - 60分钟后的20％。 另一种微创的可能性是使用激光多普勒血流计（LDF）的操作过程中，允许其成功的直接控制。在一个单个的动物，减少到10 - 20％preocclusion值明确表示局灶性脑缺血的成功归纳4然而，自卫队可以不被作为个体间的比较的方法使用，因为自卫队可以只测量定量的变化血（百分比）。流内的一个小而有限的组织样本量。它不给的血流量减少的空间范围的信息。 有几种测试，以评估中风后的行为方面，包括步态分析6,7，Rotarod 8，极测试9,10，不干胶去除测试11,12，13,14楼梯测试，梯级测试15,16和Morris水迷宫17。在所有这些测试中，受到缺血的小鼠比对照组动物执行较为成功。 <img src="/files/ftp_upload/2423/2423fig1.jpg" alt="图1" /> 图1。船只提供在小鼠大脑的架构（背景描绘）的计划。不同菌株可能会出现变化，例如的枕动脉，有时叶片颈内动脉。 <img src="/files/ftp_upload/2423/2423fig2.jpg" alt="图2" /> 图2。近端MCAO模型再灌注后不同时间点的典型病变大小的示意图。在中间，功能活动和脑血流量MCAO后的典型当然是描绘。 （MCAO：大脑中动脉闭塞，LDF激光多普勒血流测量）

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M., Prass, K., Meisel, A., Dirnagl, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Central+nervous+system+injury-induced+immune+deficiency+syndrome.">Central nervous system injury-induced immune deficiency syndrome.</a> Nat Rev Neurosci. 6, 775-786 (2005).</li><li>Engel, O., Meisel, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Models+of+Infection+Before+and+After+Stroke:+Investigating+New+Targets.">Models of Infection Before and After Stroke: Investigating New Targets.</a> Infect Disord Drug Targets. 9, (2010).</li><li>Dirnagl, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bench+to+bedside:+the+quest+for+quality+in+experimental+stroke+research.">Bench to bedside: the quest for quality in experimental stroke research.</a> J Cereb Blood Flow Metab. 26, 1465-1478 (2006).</li><li>Macleod, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Good+laboratory+practice:+preventing+introduction+of+bias+at+the+bench.">Good laboratory practice: preventing introduction of bias at the bench.</a> Stroke. 40, 50-52 (2009).</li><li>Group, S. T. A. I. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recommendations+for+standards+regarding+preclinical+neuroprotective+and+restorative+drug+development.">Recommendations for standards regarding preclinical neuroprotective and restorative drug development.</a> Stroke. 30, 2752-2758 (1999).</li></ol>

动物实验Landesamt附耳Gesundheit，柏林，德国和Soziales规定的准则和法规的规定执行。

短暂的，18,19近端MCA闭塞模型，介绍了这里的模拟缺血性中风患者最常见的类型之一 。20变量灌注时间的基础上，该模型提供了不同档次的损害，从短暂性脑缺血发作（ TIA）到大不等包括主要部件的缺血半球梗死。这使研究人员研究中风后不同的病理生理机制。 20,21 手术可以在很短的时间内，生产高度重复性病变。不过，这需要彻底控制了混杂因素。22在操作技术的细微差别可能占不同的效果梗死。23,24此外，由于脑血管解剖的差异，不同的小鼠品系表现出不同的结果 。 25,26机构温度影响，低温导致较小的病灶27和热疗，更严重的赤字的神经损伤 ，28因此，温度控制和维护在这个模型是高度相关的 。29此外，血压和血气结果的重要混杂因素和需要30,31快速，微创方法的使用（非侵入性血压测量，适合和容易访问的采血网站）进行监测。建议。麻醉剂的选择也是非常重要的，因为有些人可能有保护作用，和/或血管扩张剂，例如异氟醚 32因此，接触麻醉应尽可能和规范化。我们排除经历了手术时间超过15分钟的动物。 剃须手术部位产生microabrasions和炎症，并释放头发片段。这可能会进一步加重炎症和促进当地的感染，这可能会影响中风的病理生理。住房条件，特别是使用浓缩，可能会影响中风的结果，并应标准化，并在研究报告中描述的。6,33,34使用环境的丰富和重复性其效果是一个讨论的问题。35另一个重要混杂因素是中风引起的感染的风险，特别是经过较长时间的缺血36的，这会导致额外的发病率和死亡率增加。37,38感染症状在3至5天左右，这已长期在这种模型的后续行动的重要后果。 产生结果，为发展新的治疗中风，标准化和质量控制策略有关，是非常重要的翻译中风在临床前研究39“良好实验室规范 ”40,41任务。 <ol><li>使用动物的适当和详细的描述; </li><li>样本量的计算和报告预期的效果大小; </li><li>纳入和排除标准，之前设置的研究; </li><li>随机的分配到组; </li><li>分配隐蔽性调查; </li><li>报告从分析中排除的动物; </li><li>盲法评估的结果; </li><li>报告潜在的利益冲突和研究经费。 </li></ol>

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		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Surgical scissors (skin cut)</td>
<td>&nbsp;</td>
<td>FST</td>
<td>14028-10</td>
<td>&nbsp;</td>
<tr>
<td>2 Dumont forceps (Medical #5 straight tip)</td>
<td>&nbsp;</td>
<td>FST</td>
<td>11253-20</td>
<td>&nbsp;</td>
<tr>
<td>Spring scissors (according to Vannas)</td>
<td>&nbsp;</td>
<td>FST</td>
<td>15000-00</td>
<td>&nbsp;</td>
<tr>
<td>Applying forceps for micro clamps</td>
<td>&nbsp;</td>
<td>FST</td>
<td>00072-14</td>
<td>&nbsp;</td>
<tr>
<td>Micro vascular clamp (e.g. S&amp;G B1-V)</td>
<td>&nbsp;</td>
<td>FST</td>
<td>00396-01</td>
<td>Also Serrefine possible</td>
</tr>
<tr>
<td>2 Hemostats according to Hartmann</td>
<td>&nbsp;</td>
<td>FST</td>
<td>13002-10</td>
<td>&nbsp;</td>
<tr>
<td>Needle holder (according Olsen-Hegar or other)</td>
<td>&nbsp;</td>
<td>FST</td>
<td>12002-14</td>
<td>&nbsp;</td>
<tr>
<td>Standard anatomical forceps (for wound closure)</td>
<td>&nbsp;</td>
<td>FST</td>
<td>11000-12</td>
<td>&nbsp;</td>
<tr>
<td>5/0 sutures </td>
<td>&nbsp;</td>
<td>Suprama</td>
<td>&nbsp;</td>
<td>for vascular ligatures</td>
</tr>
<tr>
<td>6/0 sutures </td>
<td>&nbsp;</td>
<td>Suprama</td>
<td>&nbsp;</td>
<td>for skin suture, 5/0 also possible</td>
</tr>
<tr>
<td>Lidocaine (e.g. Xylocain Gel 2%)</td>
<td>&nbsp;</td>
<td>Astra Zeneca</td>
<td>&nbsp;</td>
<td>or other local pain relief</td>
</tr>
<tr>
<td>Dissecting microscope (stereo microscope), magnification 6x to max. 40x</td>
<td>&nbsp;</td>
<td>Leica Zeiss</td>
<td>MZ6 Stemi2000C</td>
<td>&nbsp;</td>
<tr>
<td>Cold light source</td>
<td>&nbsp;</td>
<td>Leica</td>
<td>KL1500</td>
<td>&nbsp;</td>
<tr>
<td>Temperature feedback controlled heating pad system</td>
<td>&nbsp;</td>
<td>FST</td>
<td>21052-00</td>
<td>With mouse pad and small probe</td>
</tr>
<tr>
<td>Anaethesia system for isoflurane</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Isoflurane</td>
<td>&nbsp;</td>
<td>Abott</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Veterinary Recovery Chamber</td>
<td>&nbsp;</td>
<td>Peco Services</td>
<td>V1200</td>
<td>Heated cage</td>
</tr>
<tr>
<td>8.0 nylon filament</td>
<td>&nbsp;</td>
<td>Suprama</td>
<td>TEL181005</td>
<td>for coating</td>
</tr>
<tr>
<td>Scalpel</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>For cutting filament in pieces</td>
</tr>
<tr>
<td>Ruler</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>To cut correct length of filament</td>
</tr>
<tr>
<td>Xantopren M Mucosa</td>
<td>&nbsp;</td>
<td>Heraeus Kulzer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Activator Universal Plus</td>
<td>&nbsp;</td>
<td>Heraeus Kulzer</td>
<td>&nbsp;</td>
<td>Optosil - Xantopren</td>
</tr>
<tr>
<td>2 Dumont forceps (Medical #5 straight tip)</td>
<td>&nbsp;</td>
<td>FST</td>
<td>11253-20</td>
<td>&nbsp;</td>
<tr>
<td>A soft underlay for storing and grasping the uncoated filaments</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>e.g. swabs, foam, &#x2026;</td>
</tr>
<tr>
<td>Receptacle for new build filaments</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>e.g. petri dish, flexible foam,&#x2026;</td>
</tr>		</tbody>
		</table>
		</div>

modeling stroke in mice - middle cerebral artery occlusion with the filament model

中风是之间的死亡和残疾成人，尤其是在高度发达的国家，最常见的原因。然而，迄今为止的治疗方案是非常有限的的。为了满足新的治疗方法需要，中风的实验研究经常采用局灶性脑缺血啮齿动物模型。大多数研究人员使用的小鼠或大鼠大脑中动脉（MCA）的永久或暂时闭塞。 通过腔内缝合技术（所谓长丝或缝合模型）（MCA）的大脑中动脉近端闭塞可能是实验性中风的研究中最常用的模型。腔内MCAO模型提供了在一个相对非侵入性的方式重现短暂或永久MCA的领土缺血诱导的优势。腔内的方法中断的整个领土的这个动脉的血流量。因此，长丝闭塞逮捕流近端lenticulo纹动脉，供应基底节。长丝闭塞，在皮层和纹状体的重现性病变MCA的结果可以是永久或短暂。相比之下，MCA闭塞模型，诱导远端（lenticulo纹动脉分支）通常备用纹状体和主要涉及的大脑皮层。此外，这些模型需要去骨瓣。本文演示模型，硅涂层长丝引入左颈总动脉和先进的威利斯，它阻断大脑中动脉的起源圈沿颈内动脉。大脑中动脉闭塞的患者，是缺血性中风的最常见的原因。由于不同缺血时间间隔可自由选择在这个模型再灌注时间点上，可以产生不同程度的缺血性病变。再灌注去除咬合长丝至少部分模型自发或治疗（TPA）在人类的血栓栓塞血块溶解后恢复血液流动。 在这个视频中，我们将目前的基本技术以及主要缺陷和混杂因素，这可能会限制这个模型的预测值。

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Modeling Stroke in Mice - Middle Cerebral Artery Occlusion with the Filament Model

丝状的大脑中动脉闭塞，是一个研究小鼠缺血性中风的常见模型。

建模在小鼠体内的行程 - 中东与灯丝模型大脑中动脉闭塞

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are ...

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90.9K 次观看.  Center for Stroke Research Berlin, Charité Universitätsmedizin. 丝状的大脑中动脉闭塞，是一个研究小鼠缺血性中风的常见模型。