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>

<ol>
	<li>Overgaard, K., Sereghy, T., Boysen, G., Pedersen, H., H&#248;yer, S., Diemer, N. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rat+model+of+reproducible+cerebral+infarction+using+thrombotic+blood+clot+emboli.">A rat model of reproducible cerebral infarction using thrombotic blood clot emboli.</a> J Cereb Blood Flow Metab. 12 (3), 484-490 (1992).</li><li>Zhang, R. L., Chopp, M., Zhang, Z. G., Jiang, Q., Ewing, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rat+model+of+focal+embolic+cerebral+ischemia.">A rat model of focal embolic cerebral ischemia.</a> Brain Res. 766 (1-2), 83-92 (1997).</li><li>Dinapoli, V. A., Rosen, C. L., Nagamine, T., Crocco, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+MCA+occlusion:+a+precise+embolic+stroke+model.">Selective MCA occlusion: a precise embolic stroke model.</a> J. Neurosci. Methods. 154 (1-2), 233-238 (2006).</li><li>Jespersen, B., Knupp, L., Northcott, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Femoral+arterial+and+venous+catheterization+for+blood+sampling,+drug+administration+and+conscious+blood+pressure+and+heart+rate+measurements.">Femoral arterial and venous catheterization for blood sampling, drug administration and conscious blood pressure and heart rate measurements.</a> J Vis Exp. 59 (3496), (2012).</li><li>Bederson, J. B., Pitts, L. H., Tsuji, M., Nishimura, M. C., Davis, R. L., Bartkowski, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+middle+cerebral+artery+occlusion:+evaluation+of+the+model+and+development+of+a+neurologic+examination.">Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination.</a> Stroke. 17 (3), 472-476 (1986).</li><li>Henninger, N., Bouley, J., Br&#229;tane, B. T., Bastan, B., Shea, M., Fisher, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser+Doppler+flowmetry+predicts+occlusion+but+not+tPA-mediated+reperfusion+success+after+rat+embolic+stroke.">Laser Doppler flowmetry predicts occlusion but not tPA-mediated reperfusion success after rat embolic stroke.</a> Exp Neurol. 215 (2), 290-297 (2009).</li><li>Zhu, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Annexin+A2+combined+with+low-dose+tPA+improves+thrombolytic+therapy+in+a+rat+model+of+focal+embolic+stroke.">Annexin A2 combined with low-dose tPA improves thrombolytic therapy in a rat model of focal embolic stroke.</a> J Cereb Blood Flow Metab. 30 (6), 1137-1146 (2010).</li><li>Cheng, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activated+protein+C+inhibits+tissue+plasminogen+activator-induced+brain+hemorrhage.">Activated protein C inhibits tissue plasminogen activator-induced brain hemorrhage.</a> Nat Med. 2 (11), 1278-1285 (2006).</li><li>Taniguchi, H., Andreasson, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+hypoxic-ischemic+encephalopathy+model+of+perinatal+ischemia.">The hypoxic-ischemic encephalopathy model of perinatal ischemia.</a> J Vis Exp. 21 (955), (2008).</li><li>Sicard, K. M., Fisher, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+focal+brain+ischemia.">Animal models of focal brain ischemia.</a> Exp Transl Stroke Med. 1 (7), (2009).</li><li>Florian, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+hypothermia+reduces+infarct+volume+in+aged+rats+after+focal+ischemia.">Long-term hypothermia reduces infarct volume in aged rats after focal ischemia.</a> Neurosci Lett. 438 (2), 180-185 (2008).</li><li>Noor, R., Wang, C. X., Shuaib, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+hyperthermia+on+infarct+volume+in+focal+embolic+model+of+cerebral+ischemia+in+rats.">Effects of hyperthermia on infarct volume in focal embolic model of cerebral ischemia in rats.</a> Neurosci Lett. 349 (2), 130-132 (2003).</li><li>Westendorp, W. F., Nederkoorn, P. J., Vermeij, J. D., Dijkgraaf, M. G., van de Beek, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Post-stroke+infection:+a+systematic+review+and+meta-analysis.">Post-stroke infection: a systematic review and meta-analysis.</a> BMC Neurol. 11 (110), (2011).</li></ol>

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，稳定性及递交难以管理，这导致变化的梗塞面积和受影响的大脑区域2,3。以往的研究表明，可重复的缺血性病变，在马华领土时，阻碍血栓被卡在马华2,3的干，就必须做到。为了准确地提出了血块和产生一个可重复栓塞性中风的模式，在这项研究中，我们将演示如何准备改性PE-50管，以及如何修改的管尖介绍给马华的起源，以及如何为纤维蛋白凝块丰富注入马华。一致的与前一报告2，注入的血液凝块被容易地在MCA的所有盐水处理的大鼠的原点可视化（N 9 =），但主要是溶解在所有的tPA治疗的大鼠组（n = 12）在24小时后栓塞。 来评价MCAO的成功，该脑血流量，神经功能障碍，并且图案和脑损伤的分布进行评估。血块注射后，脑血流量减少至基线的30％，这个减小的rCBF的持续至少2小时，栓塞后，与先前的报道6,7一致。使用tPA治疗，在栓塞后2小时后，脑血流量水平的tPA治疗30分钟后恢复到接近基准线。神经功能评分，测定10分钟之前使用修改后的Bederson得分帮助评估缺血的成功戒毒。这个神经系统的得分是一个简单而快速的方法来检测全球神经功能在中风急性期。动物显示出正常的行为（得分= 0）被排除在药物治疗和进一步分析。此外，我们证明了组织病变产生多为内马华领土在新皮层和纹状体区域看到，和TPA处理（在2小时）显著缩小梗死面积，在中风后24小时。总之，我们的数据显示，在这项工作中提出的栓塞性中风模型可以开发马华境内可预见的梗死体积。 最后，我们注意到，有几个技术问题，可能会影响栓塞MCAO模型的成功。在执行栓塞缺血模型中遇到的一个常见问题是栓塞后早期自然灌注。自发性再灌注í的次数那么很有可能要与脆弱血管外凝结的血块和用于闭塞MCA 2,3,10凝块的长度有关。我们相信，我们描述制备血液凝块的方法（步骤2）将最小化的脆弱血管外凝结的血块。单血块用于MCA闭塞的长度从实验室到不同实验室之间25毫米至50毫米的长度2,3,6-8,10。我们发现，使用了35-40毫米长的血栓闭塞理想马华和生产的高重现性梗死体积。修改PE-50管，0.30-0.34毫米之间的尖端直径。如果尖端直径&gt;0.34毫米，它不能达到的MCA的来源。如果尖端直径为&lt;0.30毫米，改性PE-50管内的凝块是难以通过尖。死亡率密切相关，严重脑肿胀，脑出血。在这项工作中，24小时死亡率为16％（4月25日，见代表性的结果），和所有死老鼠表现出严重的脑肿胀荷兰国际集团虽然我们没有看到脑出血（可能是由于样本量有限）。可控死亡率的因素是手术过程的体温。在控制从手术开始至36.5-37.5℃的体温，直到从麻醉中完全恢复。体温显著影响的脑组织损伤的程度。体温降低，体温过高会增加脑梗死体积11,12。在进行外科手术中，根据以产生高度可重复的梗死面积的协议以短时间（大约20-25分钟）。感染性并发症是导致死亡的主要原因在急性脑卒中患者13。以减少感染并发症的发生率和提高中风后的存活率，无菌技术应该在手术过程中被使用。 总之:在这篇文章中介绍的技术将有助于研究人员克服了建立个技术问题是型号为中风研究。

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 border="0" cellpadding="0" cellspacing="0" width="739">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:232px;">rh-tPA</td>
			<td style="width:135px;">Chemical</td>
			<td style="width:237px;">Genentech</td>
			<td style="width:135px;"></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:232px;">2,3,5-triphenyltetrazolium chloride</td>
			<td style="width:135px;">Chemical</td>
			<td style="width:237px;">Sigma</td>
			<td style="width:135px;">T8877</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:232px;">Anesthesia vaporizer</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">Soma Technology</td>
			<td style="width:135px;">Drager Vapor 19.1&#160;&#160;</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:232px;">Rechargeable high speed micro drill</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">Fine Science Tools</td>
			<td style="width:135px;">18000-17</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Curved scissors</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">Fine Science Tools</td>
			<td style="width:135px;">14117-14</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:232px;">Dumont forceps(Medical #7)</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">Fine Science Tools</td>
			<td style="width:135px;">11270-20</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:232px;">Dumont forceps(Medical #5)</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">Fine Science Tools</td>
			<td style="width:135px;">11251-35</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:232px;">Vannas-style spring scissors</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">Fine Science Tools</td>
			<td>15000-03</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:232px;">Veterinary recovery chamber&#160;</td>
			<td style="width:135px;">Equipment&#160;</td>
			<td style="width:237px;">Peco Services&#160;</td>
			<td style="width:135px;">V1200&#160;&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:232px;">Genie plus syringe pump</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">Kent Scientific Corporation</td>
			<td style="width:135px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:232px;">Rat brain matrix</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">Kent Scientific Corporation</td>
			<td style="width:135px;">RBMA-310C</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:232px;">Digital caliper&#160;</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">World Precision Instruments</td>
			<td style="width:135px;">501601</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Dissecting microscope</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">World Precision Instruments</td>
			<td style="width:135px;">PZMTIII-BS-LWD</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:232px;">Hamilton syringe</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">Hamilton</td>
			<td style="width:135px;">model 710</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:232px;">Homeothermic blanket control unit</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">Harvard Apparatus</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:232px;">Electric clipper</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">Braintree Scientific</td>
			<td style="width:135px;">CLP-9931</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:232px;">Veterinary electrosurgical unit</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">MACAN Manufacturing Company</td>
			<td style="width:135px;">MV-9</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:232px;">Blood flowmeter</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">Adinstruments</td>
			<td style="width:135px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:232px;">PowerLab 4/30</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">Adinstruments</td>
			<td style="width:135px;"></td>
		</tr>
		<tr height="18">
			<td height="18" style="height:18px;width:232px;">LabChart 7.2</td>
			<td style="width:135px;">software</td>
			<td style="width:237px;">Adinstruments</td>
			<td style="width:135px;"></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:232px;">1ml syringe</td>
			<td style="width:135px;">Consumable</td>
			<td style="width:237px;">Becton, Dickinson and Company</td>
			<td style="width:135px;">309659</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:232px;">23G needle</td>
			<td style="width:135px;">Consumable</td>
			<td style="width:237px;">Becton, Dickinson and Company</td>
			<td style="width:135px;">305143</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:232px;">30G needle</td>
			<td style="width:135px;">Consumable</td>
			<td style="width:237px;">Becton, Dickinson and Company</td>
			<td style="width:135px;">305106</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:232px;">PE-50 tubing</td>
			<td style="width:135px;">Consumable</td>
			<td style="width:237px;">Becton, Dickinson and Company</td>
			<td style="width:135px;">427517</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:232px;">PE-10 tubing</td>
			<td style="width:135px;">Consumable</td>
			<td style="width:237px;">Becton, Dickinson and Company</td>
			<td style="width:135px;">427400</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">6-0 Silk suture</td>
			<td style="width:135px;">Consumable</td>
			<td>Harvard apparatus</td>
			<td style="width:135px;">723287</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:232px;">5-0 Silk suture</td>
			<td style="width:135px;">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少动物&gt;基线的70％被认为是马华的成功栓塞性闭塞（ 图5）。在注射血块后2小时，太平资产（10毫克/千克）7,8的标准大鼠剂量使用注射输液泵7,8静脉注射了10％，并推注90％持续输注30分钟。我们观察到，局部脑血流量水平逐步increas版内的tPA治疗的30分钟的基线的&gt; 70％（ 图5）。 在栓塞后24小时，将动物安乐死，transcardially灌注用200ml PBS中以除去血管内的血液。以往的研究2,3,7,8和我们的数据（ 图6）显示，在栓塞后24小时产生大得多的组织梗死。大脑收集并合影留念。在生理盐水处理组中，血液凝块被容易地可视化在MCA和ACA的起源，但是血块几乎完全溶解在tPA的治疗组（ 图6A）。在这之后，大脑切成7与冰大鼠脑基质2毫米冠状切片。孵育在2％2,3,5 -氯化三苯基四氮唑（TTC）的脑切片，在37℃下进行30分钟9,10。 TTC染色后，在冠状切片被放置在平板上并拍照（ 图6B）。 inf文件的区域<html> arction在每个切片是由图像分析系统（医学图像的全国学院）测定，平均梗塞体积进行了计算和乘法部之间的距离。这栓塞性中风模型制作马华境内组织梗死作为新皮层和纹状体区域（ 图6B）所示。早期再灌注，建立了tPA的静脉给药，在栓塞后2小时，脑梗死体积均与生理盐水治疗组（394.2±68.2毫米相比的tPA治疗组（229.1±45.7毫米3，N = 12）显著减少3，N = 9），（P &lt;0.01）。 24小时死亡率为16％（4月25日）。 <img alt="图1" src="/files/ftp_upload/51956/51956fig1highres.jpg" /> 图1的测量用数字游标卡尺的改性PE-50管的外径。 NT"FO:保together.within页="总是"&gt; <img alt="图2" src="/files/ftp_upload/51956/51956fig2highres.jpg" /> 图2洗涤血液凝块在PE-10管的血液凝块洗涤通过替代推动两个注射器相连的PE-50管的端部。 <img alt="图3" fo:content-width="5in" src="/files/ftp_upload/51956/51956fig3highres.jpg" width="600" /> 图3（一）简化大鼠计划右脑动脉孤立出连续缝合，以准备采用的改性PE-50管。在大鼠大脑中动脉架构（B）计划，并从非洲经委会以先进的导管ICA的大鼠。一个单一的血液凝块（黑色）中的包含在管。 <img alt="图4" src="/files/ftp_upload/51956/51956fig4highres.jpg" /> 图4卜位于为2mm后方和5mm RR孔（直径为1.5mm）横向于前囟。 <img alt="图5" src="/files/ftp_upload/51956/51956fig5highres.jpg" /> 图5局部脑血流量（rCBF），使用激光多普勒血流仪（LDF）测定。血栓注射导致&gt; 70％脑血流量减少的基线值。 tPA的（10毫克/千克）在凝块喷射恢复的rCBF接近基线的tPA治疗的30分钟后2小时后处理。数据表示为平均值±标准差。 <img alt="图6" src="/files/ftp_upload/51956/51956fig6highres.jpg" /> 图6（A）中示出的血液凝块（黑色箭头），在大脑中动脉（MCA）的原点，并在24小时中风后大脑前动脉（ACA）的代表性图片。左，盐水处理的大鼠;右，tPA的治疗的大鼠，酒吧= 5毫米。 <strong&gt;的（b）显示，在中风后24小时TTC染色脑切片代表性的图像。左，盐水处理的大鼠;右，tPA的治疗的大鼠，酒吧= 5毫米。

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

Nanyang Technological University

Research

JoVE Journal

Medicine

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

26.9K 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

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

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are very limited. To meet the need for novel therapeutic approaches, experimental stroke research frequently employs rodent models of focal cerebral ischaemia. Most researchers use permanent or transient occlusion of the middle cerebral artery (MCA) in mice or rats.
Proximal occlusion of the middle cerebral artery (MCA) via the intraluminal suture technique (so called filament or suture model) is probably the most frequently used model in experimental stroke research. The intraluminal MCAO model offers the advantage of inducing reproducible transient or permanent ischaemia of the MCA territory in a relatively non-invasive manner. Intraluminal approaches interrupt the blood flow of the entire territory of this artery. Filament occlusion thus arrests flow proximal to the lenticulo-striate arteries, which supply the basal ganglia. Filament occlusion of the MCA results in reproducible lesions in the cortex and striatum and can be either permanent or transient. In contrast, models inducing distal (to the branching of the lenticulo-striate arteries) MCA occlusion typically spare the striatum and primarily involve the neocortex. In addition these models do require craniectomy. In the model demonstrated in this article, a silicon coated filament is introduced into the common carotid artery and advanced along the internal carotid artery into the Circle of Willis, where it blocks the origin of the middle cerebral artery. In patients, occlusions of the middle cerebral artery are among the most common causes of ischaemic stroke. Since varying ischemic intervals can be chosen freely in this model depending on the time point of reperfusion, ischaemic lesions with varying degrees of severity can be produced. Reperfusion by removal of the occluding filament at least partially models the restoration of blood flow after spontaneous or therapeutic (tPA) lysis of a thromboembolic clot in humans.
In this video we will present the basic technique as well as the major pitfalls and confounders which may limit the predictive value of this model.

Filamentous occlusion of the Middle cerebral artery is a common model for studying ischemic stroke in mice.

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 ...

科研

医学

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 ...

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 ...

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 ...

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 ...

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 ...

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, ...