作者感谢劳里·冯·Melchner，伯尔尼大学医院的神经外科，伯尔尼，瑞士，校对和编辑稿件和Paskus耶利米，波士顿儿童医院，波士顿校对的初稿。我们赞赏动物护理，麻醉，并从丹尼尔·梅特勒，数字电压表，缪勒，数字电压表，丹尼尔Zalokar和Olgica Beslac，实验外科研究所临床研究部，伯尔尼大学，瑞士伯尔尼手术援助熟练的管理。我们感谢迈克尔·伦施，研究部主管护士，重症监护医学系，伯尔尼大学医院和伯尔尼大学，瑞士伯尔尼，对于生理参数的实时数据监控和后期处理。我们感谢艾丁Nevzati，卡尔·室井和莎乐美ERHARDT，其优秀的实验室技术和手术援助。 这项工作得到了强化班部支持ê病急救医学，伯尔尼大学医院和伯尔尼，瑞士伯尔尼，临床研究部，伯尔尼，瑞士伯尔尼大学的大学，并从Kantonsspital阿劳，阿劳，瑞士的研究基金。我们感谢爱思唯尔，对于图1和2转载的许可。

三个月大的雌性新西兰兔体重2.5 - 3.5千克的用于该过程。按照卫生指引，国家研究院​​实验动物的管理和使用，并与瑞士伯尔尼（批准105＃/ 13）的广的动物护理委员会的批准进行这项研究。所有的手术在无菌条件下，在临床研究部的实验外科研究所在伯尔尼大学医院在伯尔尼，瑞士被执行。兽医麻醉医师在手术和恢复整个监测的动物。 1，动物准备，定位和锁骨下动脉插管<ol><li>诱导全身麻醉的兔肌内注射氯胺酮（30毫克/千克;的Ketalar，50毫克/毫升）和甲苯噻嗪（6毫克/公斤; Xylapan 20毫克/毫升）和麻醉的控制深度通过检查兔子的响应有毒stimulat离子（ 如脚趾捏）。看到1.7的情况下积极响应。 </li><li>拉下两只眼睛的下眼睑，并涂抹少量药膏给眼皮，防止干燥和进一步的刺激。 </li><li>导尿侧耳静脉用20 Gbutterfly venflon（20克静脉插管），用胶布固定，并连接到含有0.9％氯化钠重力袋（500毫升/ 24小时）和氯胺酮（40毫克/千克/小时） /甲苯噻嗪（4毫克/千克/小时）的连续静脉注射（IV）麻醉。管理额外的止痛药，每15分钟静脉注射（芬太尼，1微克/千克）。注意:避免挥发性气体麻醉剂，其具有降低的CPP关联，增加脑血流量，降低脑代谢速率为氧气11静脉注射麻醉剂通过维持脑血流和脑血管收缩，12研究脑时这是最上层的重要性提供了神经外科麻醉更理想的特性。血管痉挛。另外，虽然死亡率在SPONT增加aneously呼吸的动物，它可以更好地模仿急性蛛网膜下腔出血的人的情况。 </li><li>提供氧气（1 - 2升/分钟）通过呼吸面罩，使潮气末二氧化碳（ETCO 2）监控。 </li><li>在兔子的腹侧面的三角排列安装一个3通道的心电图（ECG）.Place 3皮下电极;具体地，将一个电极在右midthoracic区域（用距离到无菌剃字段用于锁骨下动脉插管）和两个电极中的分布在两肢的下腹部。 </li><li>通过下面的呼吸频率，从所述ECG信号监测心脏速率（HR），而反应对伤害性刺激监测麻醉深度的手术过程中，每隔15分钟。 </li><li>如果对伤害性刺激（趾捏）积极响应，通过推注氯胺酮（6毫克/千克）和静脉推注甲苯噻嗪（0.05毫克/千克），IV和/或额外的镇痛推注芬太尼（1 MC调整麻醉深度克/公斤）四</li><li>修复兔中对身体加温板仰卧位置，倾斜头部20°下，使之略微对侧的侧面上的锁骨下动脉将被暴露。 </li><li>申请眼膏和剃头发上绕锁骨的中间三分之一的右胸肌准备区进行手术，并在frontal-，parietal-和枕骨，颈部，并在右股总动脉。 </li><li>皮肤消毒3分钟与广谱防腐剂， 如 ，聚维酮碘。 </li><li>盖兔用无菌床单。进行无菌条件下所有进一步的程序和频繁应用4％罂粟碱盐酸和抗生素溶液（硫酸新霉素5毫克/毫升）局部给药，以防止动脉血管痉挛由船只操纵和局部感染。 </li><li>渗入胸肌与局部麻醉剂（1％利多卡因最大6毫克/千克）。使胸骨旁切开皮肤和准备胸肌。使用显微镜，解剖锁骨下动脉和近端和远端结扎（4-0 polyfilament缝合线）在外露的末端固定。保留一条绑带靠近近端控制到位，以固定导管和结扎血管远侧。 </li><li>通过切割与弯曲microscissor动脉执行在锁骨下动脉的壁上的动脉切开术和导管插入锁骨下动脉逆行用小血管内三通活栓。固定导管由面向远端结扎双结结扎，以防止动脉扭曲或动脉的近侧部分的弯曲，并避免滑动或大​​量出血。 </li></ol> 2，血压和动脉血液气体监测<ol><li>连接3通止水栓，以便i）的血管内插管用于动脉血气（ABG）分析，pH值， 二氧化碳分压， 氧分压 ，碳酸氢盐，碱过量和SO 2，ⅱ）侵入动脉血压力测量装置，以及iii）在分流装置。 </li><li>收集血液样品用于ABG状态（ 二氧化碳分压， 氧分压 ）和连续监测标准心血管和呼吸参数（血压，心率，心电图，呼吸率和潮气末CO 2）和传输经模拟输出接口数据到的模拟电源数字转换器/数据记录器和存储。 注:压力会在心脏水平在每次会议后归零，该系列开始之前的模拟/数字转换器和数据记录系统的压力校准会做一次。 </li></ol> 3，基准线数字减影血管造影<ol><li>放置一个外部尺寸测量装置（小球）上既下颚角以校准血管造影。 注:这允许事后测量基线的精确比较及跟进血管直径。 </li><li>逆行内-A进行数字减影血管造影（DSA）rterial弹丸注射非离子型碘帕醇（0.6毫升/千克，为5毫升/秒，2秒）通过管状动脉，并为了防止后者的闭塞弹丸注射用生理盐水后立即冲洗套管。 </li><li>在5°左斜前方位置获得使用快速连续造影记录的椎基底动脉系统的图像（14秒7的图像）。 </li><li>渗透到右股总动脉与局部麻醉剂（利多卡因1％，最大6毫克/千克）周围的区域。做一个小腹股沟皮肤切口。使用显微镜进行可视化，解剖股总动脉和近端和远端结扎（4-0 polyfilament缝合线）在外露的末端固定。 </li><li>动脉切开后，导管插入股动脉与5-F护套。冲洗用生理盐水护套的侧端口。 </li><li>提前5-F导管插入通过透视下的套头臂动脉。创建一个路线图，然后推进导丝为thê椎基底动脉系统。注入非离子型碘帕醇的丸（0.6毫升/千克，为5毫升/秒2秒）的DSA的步骤3.2所述。 </li></ol> 4，旋转俯卧位<ol><li>以下基准DSA，重新兔子从仰卧到俯卧位。要小心，不要操作或放置动脉导管的位置。 </li><li>定位头的头架，在30°角，面向低着头。 </li></ol> 5，小脑延髓池穿刺<ol><li>皮肤消毒过的头部和颈部用聚维酮碘3次，每次1分钟，并盖上无菌片的外科手术区域。 </li><li>插入一个22克×40mm的小儿脊柱接近针经皮枕大池没有任何事先切开皮肤或肌肉移位。 </li><li>确认该动物是通过确保缺少脚趾捏响应沿骨枕外隆凸滑针下来才完全麻醉直到检测到一个间隙;针任何进一步的不推。 </li><li>通过观察脑脊液与兔子的头部倾斜向下以20°角为几分钟自发滴落确认针的正确位置。 </li></ol> 6，安装颅内压和脑血流监测<ol><li>后正中线皮肤和帽状切口，插入一个小的手术牵开。 </li><li>使三个圆形截骨（2毫米直径）使用根据颅骨外标头骨的额骨部分高速微型钻头（ 图1）9， 即在嗅球和双侧额叶的神经监测设备，如果放置有必要的。用毫米刻度标尺来确定钻孔位置的坐标如下:颅内压（ICP）监测的midpupillary线，一条从正中矢状线两条毫米;脑实质激光多普勒探头四点五十六mm前部和侧到前囟门（ 图1）。 </li><li>可视化的硬脑膜，并进行细致的止血:用骨蜡止血骨凭借其填塞作用，并使用硬脑膜双极电凝进行局部止血。 </li><li>将脑实质的颅内压（ICP）的显示器前端插入右侧嗅球至2mm的深度，然后进行校准。 </li><li>使用外部夹紧系统放置在两个激光多普勒血流仪微细针的探头，并将它们插入对应的毛刺的孔在两个左，右额叶脑半球外侧到脑室系统， 即 ，在正中线，以避免与脑脊液干扰。放置针探针，以2.5mm的深度。 </li><li>的神经监测探针放置后，密封所有毛刺的孔用厚插头骨蜡，以保持所述颅流体密封。 </li><li>测量的平均动脉血压的基准参数（MAP），ICP和使用多参数监测和四通道激光多普勒组织血流灌注监测脑血流量（CBF）。 </li></ol> 7，分流诱导<ol><li>在小脑延髓池连接脊椎接入针经血液充满压力监测管先前插管锁骨下动脉。使用上述活栓的血压测量和血液取样口3路。 注:蛛网膜下腔出血的严重程度取决于血液的量，并且可以通过蛛网膜下腔血块在脑收获5,11时的广泛性粗略估计。 </li><li>连续监测血压，心率，心电图，呼吸频率和潮气末CO 2在1赫兹的从6分钟之前的采样率，直到SAH后至少20分钟。 </li><li>确认该动物是通过确保缺少脚趾捏响应打开锁骨下动脉和小脑延髓池诱发蛛网膜下腔出血的压力gradie之间的分流器连接之前完全麻醉NT。 注:（ 例如 ，在ICP的期望水平的）的对照蛛网膜下腔出血可以通过关闭分流在任何时间点来实现。 </li><li>在大约15分钟的时间内录得稳定状态值。 </li><li>粤ICP备后达到高峰，保持脊椎的访问针到位，直到ICP备返回到稳定状态接近基准值。如果ICP备高原保持超过10秒，或者ICP备自发降低，关闭分流。 </li><li>除去CBF细针探针和ICP探针，插入颅骨钻孔用骨蜡，除去所有的导管（包括锁骨下导管，由于导管操作与连续出血是与高发病率和死亡率相关，并增加感染发生率），进行严格的伤口灌溉用硫酸新霉素和缝合皮肤。 </li></ol> 8，术后管理<ol><li>该过程持续约2小时。由于半衰期氯胺酮和甲苯噻嗪，阴性特质的恢复时间的l为很短 - 大约1小时。该动物被关在恢复过程中的加热灯。不提供附加的流体。在此初始术后恢复阶段，申请丁丙诺啡0.02毫克/千克SC每8小时24小时。 </li><li>应用芬太尼透皮贴剂基质，在未来72小时释放出12.5微克/小时的动物进行有效的镇痛剃光颈部。 </li><li>不要让动物无人看管，直到它已经恢复了足够的意识来维持胸骨斜卧。 </li><li>不要将已动过手术在其他动物的公司，直到完全康复的动物。 </li></ol> 9跟进数字减影血管造影评估DCVS 3日<ol><li>如上所述执行步骤1.1到3.6。 </li><li>通过动脉内快速浓注的钠thiopenthal（40毫克/千克）安乐死的动物（硫喷妥钠，Ospedalia AG，Hünenberg，瑞士）。在情况下，组织学和immunohistochemistry是必要的，在室温下以100厘米的H 2 O的灌注压进行心内灌注固定</li></ol>

<ol>
	<li>Taylor, T. N., 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=Lifetime+cost+of+stroke+in+the+United+States.">Lifetime cost of stroke in the United States.</a> Stroke; a journal of cerebral circulation. 27, 1459-1466 (1996).</li><li>Kikkawa, Y., Kameda, K., Hirano, M., Sasaki, T., Hirano, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+feedback+regulation+of+the+receptor+activity+and+the+myofilament+Ca2++sensitivity+contributes+to+increased+vascular+reactiveness+after+subarachnoid+hemorrhage.">Impaired feedback regulation of the receptor activity and the myofilament Ca2+ sensitivity contributes to increased vascular reactiveness after subarachnoid hemorrhage.</a> Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism. 30, 1637-1650 (2010).</li><li>Marbacher, S., Fandino, J., Kitchen, N. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Standard+intracranial+in+vivo+animal+models+of+delayed+cerebral+vasospasm.">Standard intracranial in vivo animal models of delayed cerebral vasospasm.</a> British journal of neurosurgery. 24, 415-434 (2010).</li><li>Marbacher, S., Neuschmelting, V., Graupner, T., Jakob, S. M., Fandino, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevention+of+delayed+cerebral+vasospasm+by+continuous+intrathecal+infusion+of+glyceroltrinitrate+and+nimodipine+in+the+rabbit+model+in+vivo.">Prevention of delayed cerebral vasospasm by continuous intrathecal infusion of glyceroltrinitrate and nimodipine in the rabbit model in vivo.</a> Intensive care medicine. 34, 932-938 (2008).</li><li>Zhou, M. L., 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=Comparison+between+one-+and+two-hemorrhage+models+of+cerebral+vasospasm+in+rabbits.">Comparison between one- and two-hemorrhage models of cerebral vasospasm in rabbits.</a> Journal of neuroscience. 159, 318-324 (2007).</li><li>Vatter, 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=Time+course+in+the+development+of+cerebral+vasospasm+after+experimental+subarachnoid+hemorrhage:+clinical+and+neuroradiological+assessment+of+the+rat+double+hemorrhage+model.">Time course in the development of cerebral vasospasm after experimental subarachnoid hemorrhage: clinical and neuroradiological assessment of the rat double hemorrhage model.</a> Neurosurgery. 58, 1190-1197 (2006).</li><li>Lee, J. Y., Sagher, O., Keep, R., Hua, Y., Xi, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+experimental+rat+models+of+early+brain+injury+after+subarachnoid+hemorrhage.">Comparison of experimental rat models of early brain injury after subarachnoid hemorrhage.</a> Neurosurgery. 65, 331-343 (2009).</li><li>Marbacher, S., 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=A+new+rabbit+model+for+the+study+of+early+brain+injury+after+subarachnoid+hemorrhage.">A new rabbit model for the study of early brain injury after subarachnoid hemorrhage.</a> Journal of neuroscience. 208, 138-145 (2012).</li><li>Marbacher, S., 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=Outer+skull+landmark-based+coordinates+for+measurement+of+cerebral+blood+flow+and+intracranial+pressure+in+rabbits.">Outer skull landmark-based coordinates for measurement of cerebral blood flow and intracranial pressure in rabbits.</a> Journal of neuroscience methods. 201, 322-326 (2011).</li><li>Marbacher, S., 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=Extra-intracranial+blood+shunt+mimicking+aneurysm+rupture:+intracranial-pressure-controlled+rabbit+subarachnoid+hemorrhage+model.">Extra-intracranial blood shunt mimicking aneurysm rupture: intracranial-pressure-controlled rabbit subarachnoid hemorrhage model.</a> Journal of neuroscience. 191, 227-233 (2010).</li><li>Sugawara, T., Ayer, R., Jadhav, V., Zhang, J. 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+new+grading+system+evaluating+bleeding+scale+in+filament+perforation+subarachnoid+hemorrhage+rat+model.">A new grading system evaluating bleeding scale in filament perforation subarachnoid hemorrhage rat model.</a> J Neurosci Methods. 167, 327-334 (2008).</li><li>Macdonald, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delayed+neurological+deterioration+after+subarachnoid+haemorrhage.">Delayed neurological deterioration after subarachnoid haemorrhage.</a> Nature reviews. Neurology. 10, 44-58 (2014).</li><li>Zhang, Z. W., 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=Platelet-derived+growth+factor-induced+severe+and+chronic+vasoconstriction+of+cerebral+arteries:+proposed+growth+factor+explanation+of+cerebral+vasospasm.">Platelet-derived growth factor-induced severe and chronic vasoconstriction of cerebral arteries: proposed growth factor explanation of cerebral vasospasm.</a> Neurosurgery. 66, 728-735 (2010).</li><li>Laslo, A. M., Eastwood, J. D., Chen, F. X., Lee, T. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+CT+perfusion+imaging+in+subarachnoid+hemorrhage-related+vasospasm.">Dynamic CT perfusion imaging in subarachnoid hemorrhage-related vasospasm.</a> AJNR. American journal of neuroradiology. 27, 624-631 (2006).</li><li>Shao, Z., 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=Effects+of+tetramethylpyrazine+on+nitric+oxide/cGMP+signaling+after+cerebral+vasospasm+in+rabbits.">Effects of tetramethylpyrazine on nitric oxide/cGMP signaling after cerebral vasospasm in rabbits.</a> Brain research. 1361, 67-75 (2010).</li><li>Bederson, J. B., Germano, I. M., Guarino, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cortical+blood+flow+and+cerebral+perfusion+pressure+in+a+new+noncraniotomy+model+of+subarachnoid+hemorrhage+in+the+rat.">Cortical blood flow and cerebral perfusion pressure in a new noncraniotomy model of subarachnoid hemorrhage in the rat.</a> Stroke; a journal of cerebral circulation. 26, 1086-1091 (1995).</li><li>Veelken, J. A., Laing, R. J., Jakubowski, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Sheffield+model+of+subarachnoid+hemorrhage+in+rats.">The Sheffield model of subarachnoid hemorrhage in rats.</a> Stroke; a journal of cerebral circulation. 26, 1279-1283 (1995).</li><li>Zakhartchenko, V., 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=Cell-mediated+transgenesis+in+rabbits:+chimeric+and+nuclear+transfer+animals.">Cell-mediated transgenesis in rabbits: chimeric and nuclear transfer animals.</a> Biology of reproduction. 84, 229-237 (2011).</li><li>Capecchi, 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=Gene+targeting+in+mice:+functional+analysis+of+the+mammalian+genome+for+the+twenty-first+century.">Gene targeting in mice: functional analysis of the mammalian genome for the twenty-first century.</a> Nature reviews. Genetics. 6, 507-512 (2005).</li><li>Flisikowska, 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=Efficient+immunoglobulin+gene+disruption+and+targeted+replacement+in+rabbit+using+zinc+finger+nucleases.">Efficient immunoglobulin gene disruption and targeted replacement in rabbit using zinc finger nucleases.</a> PloS one. 6, e21045 (2011).</li><li>Nakajima, M., 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=Effects+of+aging+on+cerebral+vasospasm+after+subarachnoid+hemorrhage+in+rabbits.">Effects of aging on cerebral vasospasm after subarachnoid hemorrhage in rabbits.</a> Stroke. 32, 620-628 (2001).</li></ol>

None. The authors have no financial or commercial interest in any of the drugs, materials, or equipment used. No specific funding was received for this work. The authors are solely responsible for the design and conduct of the presented study and report no conflict of interest concerning the materials and methods used in this study or the findings specified in the paper. They confirm the adherence of ethical standards. The study was performed in accordance with the National Institutes of Health guidelines for the care and use of experimental animals and with the approval of the Animal Care Committee of the Canton of Bern, Switzerland (approval #109/07 and #107/09).

分路模型产生病理类似于在急性SAH 3,8,10后人类观察。它已被提出，EBI可能会加剧，维持，甚至触发DCVS 12，因此该模型可在调查两种早期和晚期DCVS阶段，包括以下SAH EBI和DCVS相互作用帮助。特别的，可重复的体内 DCVS的监测技术包括DSA 13，计算机断层扫描血管造影14，和经颅多普勒15顷兔多readibly施加比在较小的实验室动物。除了让考官独立自发蛛网膜下腔出血，该模型允许用户控制对不同严重程度（血液或ICP备量，并在脑灌注压随之变化）。 最重要的是，这种模式会导致一个非常一致的，可重复的，而病理生理适应急性蛛网膜下腔出血后的事件过程。再出血<html> ING是不存在的，比较急性蛛网膜下腔出血16,17等型号的死亡率相对较低。在该分流系统进一步的流探头的插入使出血10中SAH体积的实时评估。然而，随着大型动物的程序相关联的成本是显著大于较常用的小型实验室动物，和兔的遗传操作是令人望而却步具有挑战性的，尤其是基因上SAH从而限制了研究成果18-20。 为确保SAH本模型中，我们建议考虑以下的一般准则和关键的手术步骤的一致性和准确性: 3至四个月大的兔子相比，较旧的兔子，和ii）延伸于老年兔（20至40个月），21中的血管痉挛期间进行实验，因为i）该死亡率SAH诱导后显着降低。 帐篷"&gt;与外部设备的大小（小球）的血管造影的校准允许血管直径的评估及低变异性和精确的校准基线采用盲法和随访血管造影，测量血管的直径从三次（沿着预定的长度建议在基底动脉的使用自动测量分析软件来确定平均值（在协议中步骤3）的前端部）。 锁骨下动脉的安全的结扎线，以防止扭曲或锁骨下动脉的弯曲从仰卧重新定位，以俯卧位（在协议中的步骤4）中。 密封所有毛刺的孔用厚插头骨蜡之前，蛛网膜下腔出血诱导的，以保持所述颅流体密封和ICP，CPP的主要稳态和MAP（在协议第6步）。 保持脊椎接入针到位，直到ICP备返回到基线值。脊柱接入网元的错位edle可导致显著发病率（在协议第7步）。 EBI和DCVS，这两者在很大程度上有助于不利结果和死亡率SAH后，可以使用SAH的血液分流模型进行研究。的各个技术细节的意识，保证成功实施这一模式，并允许潜在的治疗模式蛛网膜下腔出血后遗症和筛选的评价。

蛛网膜下腔出血（SAH）是最威胁生命的病理条件下，经常导致永久性神经损伤或死亡1之一。过去的研究主要集中于迟发性脑血管痉挛（DCVS）为蛛网膜下腔出血2相关的神经功能障碍的主要病因。但是，患者在治疗血管痉挛后，从患蛛网膜下腔出血的普遍较差的临床结果，导致了研究的重点扩展到包括早期脑损伤（EBI），蛛网膜下腔出血3后的效果。两个EBI和DCVS在SAH后造成不良的临床结果的意义更多的理解是更有效的治疗策略的发展是必不可少的。 截至目前，单双自体血液注射到小脑延髓池一直是蛛网膜下腔出血诱导的DCVS 2-6的研究的标准方法。尽管在以前的研究中常用的这种模式很可能不会重现蛛网膜下腔出血相关的病理变化主要诱发EBI 7。与此相反，腔内穿孔已知产生严重的急性病理生理改变部分地模仿EBI 7的症状。 这份报告描述蛛网膜下腔出血的一种新的动物模型，旨在使这两个EBI和DCVS的调查，从而使蛛网膜下腔出血引起的病理8-10更准确的描述。随着所描述的技术，标准小脑延髓池模式适于通过连接动脉系统锁骨下动脉和小脑延髓池通过体外分流。的血流量，从而与兔子的生理及由动脉血和颅内压之间的压力梯度驱动。出血停止时脑压（ICP）等于舒张期血压和血液中的分流系统凝固。利用主机＆＃8217;的生理减少考官依赖SAH诱导，导致蛛网膜下腔出血比较一致的模型，可靠地产生两个EBI和DCVS表型3,8-10。

<table><tbody><tr><td>Equipment</td><td></td><td></td><td></td></tr><tr><td>Operation microscope</td><td>Zeiss, Jena, Germany</td><td></td><td>Zeiss, OPMI-MD surgical microscope</td></tr><tr><td>Surgical equipment</td><td>B. Braun, Germany</td><td></td><td>Forceps medical no. 5; vessel scissors 8 cm; microclip 4 mm</td></tr><tr><td>Respirator</td><td>Hugo Sachs</td><td></td><td></td></tr><tr><td>Hair clipper</td><td>3M Surgical Clipper&amp;nbsp;</td><td></td><td>Starter Kit 9667A</td></tr><tr><td>Body warm plate</td><td>FHC</td><td></td><td></td></tr><tr><td>Blood gas analyzer</td><td>Radiometer, Copenhagen, Denmark</td><td>ABL 725</td><td></td></tr><tr><td>Cardiac monitoring</td><td>Camino Multi-Parameter Monitor, Integra, Plainsboro, NJ, US</td><td>AP-05</td><td></td></tr><tr><td>Software analysis</td><td>BIOPAC Systems, Inc., Goleta, CA, USA</td><td></td><td>Biopac MP100 and acqKnowledge software,version 3.8.1</td></tr><tr><td>Software analysis</td><td>ImagePro Discovery, MediaCybernetics, Silver Spring, MD, USA</td><td></td><td>Image-Pro Plus version&amp;nbsp;</td></tr><tr><td>Angiography apparatus</td><td>DFP 2000 A-Toshiba</td><td>MIIXR0001EAA</td><td></td></tr><tr><td>ICP monitor</td><td>Camino Laboratories, San Diego, CA, USA</td><td></td><td>ICP monitor, Model 110-4B</td></tr><tr><td>Blood flow monitor</td><td>Oxford Optronix Ltd., Oxford, UK</td><td></td><td>CAL KIT microsphere solution</td></tr><tr><td>Laser-Doppler flowmetry fine needle probes</td><td>Oxford Optronix Ltd., Oxford, UK</td><td></td><td>MNP110XP, 0.48 mm diameter</td></tr><tr><td>Pressure tube</td><td>B. Braun, Germay</td><td></td><td>PE 1.0 mm &amp;times; 2.0 mm</td></tr><tr><td>Anesthesia monitor</td><td>GE Medical Systems, Switzerland&amp;nbsp;</td><td></td><td>Datex S5 Monitor</td></tr><tr><td>Material</td><td></td><td></td><td></td></tr><tr><td>20 G vascular catheter</td><td>Smiths Medical</td><td></td><td>Jelco i.v. catheter, REF 4057</td></tr><tr><td>5.5 F three-lumen central venous catheter&amp;nbsp;</td><td>Connectors, Tagelswangen, Switzerland</td><td></td><td>Silicone catheter STH-C040</td></tr><tr><td>22 G x 40 mm needle&amp;nbsp;</td><td>Emergo Group Inc., Netherlands</td><td></td><td></td></tr><tr><td>High-speed microdrill</td><td>Stryker, Solothurn, Switzerland</td><td>5400-15&amp;nbsp;</td><td></td></tr><tr><td>Bone wax</td><td>Ethicon, Johnson &amp;amp; Johnson,NJ, USA</td><td>ETHW31G</td><td></td></tr><tr><td>Bipolar forceps</td><td>Aesculap, Inc., PA, US</td><td>US349SP&amp;nbsp;</td><td></td></tr><tr><td>Ketamin</td><td></td><td></td><td>Any generic product</td></tr><tr><td>Xylazine</td><td></td><td></td><td>Any generic product</td></tr><tr><td>Buprenorphine</td><td></td><td></td><td>Any generic product</td></tr><tr><td>Fentanyl</td><td></td><td></td><td>Any generic product</td></tr><tr><td>Transdermal fentanyl matrix patches&amp;nbsp;</td><td></td><td></td><td>Any generic product</td></tr><tr><td>Lidocaine 1%&amp;nbsp;</td><td></td><td></td><td>Any generic product</td></tr><tr><td>4% papaverin HCl&amp;nbsp;</td><td></td><td></td><td>Any generic product</td></tr><tr><td>Neomycin sulfate&amp;nbsp;</td><td>Research Organics Inc., OH, USA</td><td></td><td>Any generic product</td></tr><tr><td>Povidone-iodine&amp;nbsp;</td><td></td><td></td><td>Any generic product</td></tr><tr><td>0.9% sodium chloride</td><td></td><td></td><td>Any generic product</td></tr><tr><td>Iopamidol&amp;nbsp;</td><td>Abott Laboratories, IL, USA</td><td></td><td>Any generic product</td></tr><tr><td>3-0 resorbable suture</td><td>Ethicon Inc., USA</td><td>VCP824G</td><td></td></tr><tr><td>5-0 non absorbable suture</td><td>Ethicon Inc., USA</td><td>8618G</td><td></td></tr><tr><td>4-0 polyfilament sutures</td><td>Ethicon Inc., USA</td><td>VCP284G</td><td></td></tr></tbody></table>

the rabbit blood-shunt model for the study of acute and late sequelae of subarachnoid hemorrhage: technical aspects

早期脑损伤和迟发性脑血管痉挛既有助于蛛网膜下腔出血（SAH）后的不良后果。在模拟两个条件重现性和可控性的动物模型是目前少见。因此，需要以模仿从蛛网膜下腔出血导致人体的病理生理条件的新车型。 这份报告描述了兔子的血液分流蛛网膜下腔出血模型，使颅内压（ICP）控制技术的细微差别。体外分流放置在动脉系统和蛛网膜下腔，它使检查者无关SAH在一个封闭的颅骨之间。一步一步的程序指令和必要的设备描述，以及技术方面的考虑，生产模式以最小的死亡率和发病率。需要手术成功创造这一功能强大，简单和一致ICP备控制蛛网膜下腔出血动物模型的重要细节进行了描述。

<html>蛛网膜下腔出血的兔血分流模式本报告中所述生产的海马EBI（ 图2A，B），基础皮层（ 图2A，B）和脑血管（ 图2C）的伤害，早在24小时后，显示出的特征血液分布（ 图2D）8。此外，该模型触发中度至第三天严重度DCVS的SAH诱导（ 图3）10后。死亡率为20 - 30％，因呼吸骤停或严重心动过缓急性蛛网膜下腔出血的时间。几乎所有的兔子表现出神经系统缺陷，从1天逐步恶化- 3 10在那个时间点上的动物是在完全麻醉。从技术上看，血液分流模式允许两个考官独立可控性SAH诱导（ 图4）。 <html>再1"FO:内容宽度="6英寸"WIDTH ="600"SRC ="/文件/ ftp_upload / 52132 / 52132fig1highres.jpg"/&gt; 图1:外颅骨地标CBF和ICP探针 （转载来自神经科学方法的杂志许可201:322 - 326）9示意图描述了在额叶及右侧嗅球安置实质内脑血流和ICP探头。 。脑血流量探头放置4 - 5 mm产品前，矢状窦旁的前囟门。从正中矢状线2毫米 - ICP探头以1:1的距离放置在尾，喙方向midpupillary线。脑实质内脑血流（图A和B）和ICP（面板C和D）的探针显示在T2加权的MRI矢状面和冠状面。注意它们之间的关系的心室。 CBF1启动=左额叶burrhole脑血流探测。 CBFR =右额颅骨钻孔脑血流探测。钻孔颅内压力传感器的ICP =位置。 *前囟门; **拉姆达。F ="htt​​ps://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52132/52132fig1highres.jpg"目标="_ blank将"&gt;请点击这里查看该图的放大版本。 <img alt="图2" fo:content-width="6in" width="600" src="/files/ftp_upload/52132/52132fig2highres.jpg" /> 图2:蛛网膜下腔出血后早期脑损伤 （从神经科学杂志方法208，138本文转载- 145 8和神经科学方法191的日志:227 - 233），10。 TUNEL染色显示细胞凋亡中的海马（A）和蛛网膜下腔出血动物的基础皮层（B）。神经退行性疾病是由FJB（氟玉染色）共定位DAPI染色阳性的细胞进行分析。共定位用DAPI（左栏）透露，TUNEL阳性染色（中间栏）中的局部存在于细胞核中（右栏）。打开箭头显示的DAPI阳性细胞核染色。实线箭头表示TUNEL阳性或FJB-positivE细胞。比例尺= 50微米（C）细胞凋亡和神经退行性疾病被描述24小时蛛网膜下腔出血后基底动脉内皮细胞。充满箭头表示TUNEL-凋亡阳性，肿胀，超然的内皮细胞。比例尺=50μm以下。大脑（D）总检查显示在脑的腹侧和背侧表面和基底池SAH诱导后24小时，相比于假手术动物延长血液分布。 * =基底池; * = prepontine蓄水池; *** =小脑延髓池;从围手术期ICP显示器尖端（**** = ICP探头病变）右额叶皮层病变。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52132/52132fig2highres.jpg" target="_blank">请点击这里查看该图的放大版本。</a> <img alt="图3" fo:content-width="4in" width="400" src="/files/ftp_upload/52132/52132fig3highres.jpg" /> 图3。椎基底动脉造影之前（A1）和后（A2）SAH诱导。基线血管造影（A1）示出了脊椎和基底动脉的正常血管直径。蛛网膜下腔出血感应（A2），三天后，血管造影显示基底动脉（箭头）和椎动脉弥漫性狭窄。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52132/52132fig3highres.jpg" target="_blank">请点击这里查看该图的放大版本。</a> <img alt="图4" fo:content-width="6in" width="600" src="/files/ftp_upload/52132/52132fig4highres.jpg" /> 图4。ICP控感应不同程度蛛网膜下腔出血的。兔血液分流模型的一个关键因素是控制了不同程度的蛛网膜下腔出血的严重程度，包括出血量，增加颅内压（ICP）的能力，或在脑灌注压（CPP）的降低， 图4示出了ICP followi的时间过程纳克感应蛛网膜下腔出血（箭头=开放的分流）。曲线级数依赖于兔的生理学，主要ICP和平均动脉血压之间的压力梯度。如果ICP达到一个值接近舒张期血压，在分流的流动停止。在该时间点或者ICP开始降低或ICP值保持在一个平台期。如果在高原停留时间超过10秒的分流是封闭的。控制蛛网膜下腔出血可以在任何层次ICP备通过自发血栓形成（X =封闭分流）前分流封闭进行。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52132/52132fig4highres.jpg" target="_blank">请点击这里查看这个数字的放大版本。</a>

Watch this Scientific Journal Video about The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects at JoVE.com

The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures &#8212; subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

蛛网膜下腔出血急性和后遗症的研究兔血分流型号:技术问题

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and ...

the-rabbit-blood-shunt-model-for-study-acute-late-sequelae

Universidad San Sebastian

Research

JoVE Journal

Medicine

13.6K 次观看.  University and Bern University Hospital (Inselspital). The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures — subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

视频: The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

Autologous Blood Injection to Model Spontaneous Intracerebral Hemorrhage in Mice

Investigation of the pathophysiology of injury after intracerebral
hemorrhage (ICH) requires a reproducible animal model. While ICH
accounts for 10-15% of all strokes, there remains no specific
effective therapy. The autologous blood injection model in mice
involves the stereotaxic injection of arterial blood into the basal
ganglia mimicking a spontaneous hypertensive hemorrhage in man. The
response to hemorrhage can then be studied in vivo and the
neurobehavioral deficits quantified, allowing for description of the
ensuing pathology and the testing of potential therapeutic agents. The
procedure described in this protocol uses the double injection
technique to minimize risk of blood reflux up the needle track, no
anticoagulants in the pumping system, and eliminates all dead space
and expandable tubing in the system.

The autologous blood injection model of intracerebral hemorrhage in mice described in this protocol uses the double injection technique to minimize risk of blood reflux up the needle track, no anticoagulants in the pumping system, and eliminates all dead space and expandable tubing in the system.

自体血液的注入模型小鼠自发性脑出血

Investigation of the pathophysiology of injury after intracerebral
hemorrhage (ICH) requires a reproducible animal model.  While ICH
accounts for 10-15% ...

科研

神经科学

A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage

Objective: To characterize and establish a reproducible model that demonstrates delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH) in rats, in order to identify the initiating events, pathophysiological changes and potential targets for treatment.
Methods: Twenty-eight male Sprague-Dawley rats (250 - 300 g) were arbitrarily assigned to one of two groups - SAH or saline control. Rat subarachnoid hemorrhage in the SAH group (n=15) was induced by double injection of autologous blood, 48 hr apart, into the cisterna magna. Similarly, normal saline (n=13) was injected into the cisterna magna of the saline control group. Rats were sacrificed on day five after the second blood injection and the brains were preserved for histological analysis. The degree of vasospasm was measured using sections of the basilar artery, by measuring the internal luminal cross sectional area using NIH Image-J software. The significance was tested using Tukey/Kramer's statistical analysis.
Results: After analysis of histological sections, basilar artery luminal cross sectional area were smaller in the SAH than in the saline group, consistent with cerebral vasospasm in the former group. In the SAH group, basilar artery internal area (.056 &mu;m &plusmn; 3) were significantly smaller from vasospasm five days after the second blood injection (seven days after the initial blood injection), compared to the saline control group with internal area (.069 &plusmn; 3; p=0.004). There were no mortalities from cerebral vasospasm.
Conclusion: The rat double SAH model induces a mild, survivable, basilar artery vasospasm that can be used to study the pathophysiological mechanisms of cerebral vasospasm in a small animal model. A low and acceptable mortality rate is a significant criterion to be satisfied for an ideal SAH animal model so that the mechanisms of vasospasm can be elucidated 7, 8. Further modifications of the model can be made to adjust for increased severity of vasospasm and neurological exams.

Aneurysmal subarachnoid hemorrhage (SAH) is bleeding that occurs into the subarachnoid space when an aneurysm ruptures. While the morbidity and mortality from this event has been on a decline due to improved treatment approaches, the risk of vasospasm after subarachnoid hemorrhage continues to be the same as it was several years ago. The importance of establishing a comprehensive and reproducible animal model to identify initiating events of cerebral vasospasm has been the focus of research since the first use of rats in an experimental vasospasm model in 1979 by Barry et al. Early work in rats demonstrated that a single injection of autologous blood into the cisterna magna led to acute (within minutes) but not delayed cerebral vasospasm 3, 6, 14. Here we characterize a low mortality SAH rat model that results in reproducible delayed vasospasm.

死亡率低大鼠模型的实验性蛛网膜下腔出血后迟发性脑血管痉挛评估

Objective: To characterize and establish a reproducible model that demonstrates delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH) ...

医学

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

蛛网膜下腔出血的小鼠模型

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

A Volumetric Method for Quantification of Cerebral Vasospasm in a Murine Model of Subarachnoid Hemorrhage

Subarachnoid hemorrhage (SAH) is a subtype of hemorrhagic stroke. Cerebral vasospasm that occurs in the aftermath of the bleeding is an important factor determining patient outcome and is therefore frequently taken as a study endpoint. However, in small animal studies on SAH, quantification of cerebral vasospasm is a major challenge. Here, an ex vivo method is presented that allows quantification of volumes of entire vessel segments, which can be used as an objective measure to quantify cerebral vasospasm. In a first step, endovascular casting of the cerebral vasculature is performed using a radiopaque casting agent. Then, cross-sectional imaging data are acquired by micro computed tomography. The final step involves 3-dimensional reconstruction of the virtual vascular tree, followed by an algorithm to calculate center lines and volumes of the selected vessel segments. The method resulted in a highly accurate virtual reconstruction of the cerebrovascular tree shown by a diameter-based comparison of anatomical samples with their virtual reconstructions. Compared with vessel diameters alone, the vessel volumes highlight the differences between vasospastic and non-vasospastic vessels shown in a series of SAH and sham-operated mice.

The goal of this article is to present a method that allows a 3-dimensional reconstruction of the cerebrovascular tree in mice after micro computed tomography and determination of volumes of entire vessel segments that can be used to quantify cerebral vasospasm in murine models of subarachnoid hemorrhage.

小鼠蛛网膜下腔出血模型脑血管痉挛量化的容积法

Subarachnoid hemorrhage (SAH) is a subtype of hemorrhagic stroke. Cerebral vasospasm that occurs in the aftermath of the bleeding is an important factor ...

Double Direct Injection of Blood into the Cisterna Magna as a Model of Subarachnoid Hemorrhage

Among strokes, subarachnoid hemorrhage (SAH) consecutive to the rupture of a cerebral arterial aneurysm represents 5-9% but is responsible for about 30% of the total stroke-related mortality with an important morbidity in terms of neurological outcome. A delayed cerebral vasospasm (CVS) may occur most often in association with a delayed cerebral ischemia. Different animal models of SAH are now being used including endovascular perforation and direct injection of blood into the cisterna magna or even the prechiasmatic cistern, each exhibiting distinct advantages and disadvantages. In this article, a standardized mouse model of SAH by double direct injection of determined volumes of autologous whole blood into the cisterna magna is presented. Briefly, mice were weighed and then anesthetized by isoflurane inhalation. Then, the animal was placed in a reclining position on a heated blanket maintaining a rectal temperature of 37 &#176;C and positioned in a stereotactic frame with a cervical bend of about 30&#176;. Once in place, the tip of an elongated glass micropipette filled with the homologous arterial blood taken from carotid artery of another mouse of the same age and gender (C57Bl/6J) was positioned at a right angle in contact with the atlanto-occipital membrane by means of a micromanipulator. Then 60 &#181;L of blood was injected in the cisterna magna followed by a 30&#176; downward tilt of the animal for 2 minutes. The second infusion of 30 &#181;L of blood into the cisterna magna was performed 24 h after the first one. The individual follow-up of each animal is carried out daily (careful evaluation of weight and well-being). This procedure allows a predictable and highly reproducible distribution of blood, likely accompanied by intracranial pressure elevation that can be mimicked by an equivalent injection of an artificial cerebral spinal fluid (CSF), and represents an acute to mild-model of SAH inducing low mortality.

We described in this protocol a standardized subarachnoid hemorrhage (SAH) mouse model by a double injection of autologous whole-blood into the cisterna magna. The high degree of standardization of the double-injection procedure represents a middle-to-acute model of SAH with relative safety regarding mortality.

双直接注射血液到西斯特纳麦格作为苏巴拉奇内德出血的模型

Among strokes, subarachnoid hemorrhage (SAH) consecutive to the rupture of a cerebral arterial aneurysm represents 5-9% but is responsible for about 30% ...

Analysis of Cerebral Vasospasm in a Murine Model of Subarachnoid Hemorrhage with High Frequency Transcranial Duplex Ultrasound

Cerebral vasospasm that occurs in the weeks after subarachnoid hemorrhage, a type of hemorrhagic stroke, contributes to delayed cerebral ischemia. A problem encountered in experimental studies using murine models of SAH is that methods for in vivo monitoring of cerebral vasospasm in mice are lacking. Here, we demonstrate the application of high frequency ultrasound to perform transcranial Duplex sonography examinations on mice. Using the method, the internal carotid arteries (ICA) could be identified. The blood flow velocities in the intracranial ICAs were accelerated significantly after induction of SAH, while blood flow velocities in the extracranial ICAs remained low, indicating cerebral vasospasm. In conclusion, the method demonstrated here allows functional, noninvasive in vivo monitoring of cerebral vasospasm in a murine SAH model.

The aim of this manuscript is to present a sonography-based method that allows in vivo imaging of blood flow in cerebral arteries in mice. We demonstrate its application to determine changes in blood flow velocities associated with vasospasm in murine models of subarachnoid hemorrhage (SAH).

高频颅内双体超声波的苏巴拉赫诺德出血的穆林模型脑血管痉挛分析

Cerebral vasospasm that occurs in the weeks after subarachnoid hemorrhage, a type of hemorrhagic stroke, contributes to delayed cerebral ischemia. A ...

Pre-Chiasmatic, Single Injection of Autologous Blood to Induce Experimental Subarachnoid Hemorrhage in a Rat Model

Despite advances in treatment over the last decades, subarachnoid hemorrhage (SAH) continues to carry a high burden of morbidity and mortality, largely afflicting a fairly young population. Several animal models of SAH have been developed to investigate the pathophysiological mechanisms behind SAH and to test pharmacological interventions. The pre-chiasmatic, single injection model in the rat presented in this article is an experimental model of SAH with a predetermined blood volume. Briefly, the animal is anesthetized, intubated, and kept under mechanical ventilation. Temperature is regulated with a heating pad. A catheter is placed in the tail artery, enabling continuous blood pressure measurement as well as blood sampling. The atlantooccipital membrane is incised and a catheter for pressure recording is placed in the cisterna magna to enable intracerebral pressure measurement. This catheter can also be used for intrathecal therapeutic interventions. The rat is placed in a stereotaxic frame, a burr hole is drilled anteriorly to the bregma, and a catheter is inserted through the burr hole and placed just anterior to the optic chiasm. Autologous blood (0.3 mL) is withdrawn from the tail catheter and manually injected. This results in a rise of intracerebral pressure and a decrease of cerebral blood flow. The animal is kept sedated for 30 min and given subcutaneous saline and analgesics. The animal is extubated and returned to its cage. The pre-chiasmatic model has a high reproducibility rate and limited variation between animals due to the pre-determined blood volume. It mimics SAH in humans making it a relevant model for SAH research.

Subarachnoid hemorrhage continues to carry a high burden of mortality and morbidity in man. To facilitate further research into the condition and its pathophysiology, a pre-chiasmatic, single injection model is presented.

视交叉前，单次注射自体血以诱导大鼠模型中实验性蛛网膜下腔出血

Despite advances in treatment over the last decades, subarachnoid hemorrhage (SAH) continues to carry a high burden of morbidity and mortality, largely ...

Endovascular Perforation Model for Subarachnoid Hemorrhage Combined with Magnetic Resonance Imaging (MRI)

The endovascular filament perforation model to mimic subarachnoid hemorrhage (SAH) is a commonly used model - however, the technique can cause a high mortality rate as well as an uncontrollable volume of SAH and other intracranial complications such as stroke or intracranial hemorrhage. In this protocol, a standardized SAH mouse model is presented, induced by endovascular filament perforation, combined with magnetic resonance imaging (MRI) 24 h after operation to ensure the correct bleeding site and exclude other relevant intracranial pathologies. Briefly, C57BL/6J mice are anesthetized with an intraperitoneal ketamine/xylazine (70 mg/16 mg/kg body weight) injection and placed in a supine position. After midline neck incision, the common carotid artery (CCA) and carotid bifurcation are exposed, and a 5-0 non-absorbable monofilament polypropylene suture is inserted in a retrograde fashion into the external carotid artery (ECA) and advanced into the common carotid artery. Then, the filament is invaginated into the internal carotid artery (ICA) and pushed forward to perforate the anterior cerebral artery (ACA). After recovery from surgery, mice undergo a 7.0 T MRI 24 h later. The volume of bleeding can be quantified and graded via postoperative MRI, enabling a robust experimental SAH group with the option to perform further subgroup analyses based on blood quantity.

Endovascular Perforation Model for Subarachnoid Hemorrhage Combined with Magnetic Resonance Imaging MRI

Here we present a standardized SAH mouse model, induced by endovascular filament perforation, combined with magnetic resonance imaging (MRI) 24 h after operation to ensure the correct bleeding site and exclude other relevant intracranial pathologies.

蛛网膜下腔出血结合磁共振成像（MRI）的血管内穿孔模型

The endovascular filament perforation model to mimic subarachnoid hemorrhage (SAH) is a commonly used model - however, the technique can cause a high ...

Establishing a Subarachnoid Hemorrhage Rat Model Induced by Autologous Blood Injection

To place the laser Doppler probe, make a 15-millimeter incision caudally in the midline, starting just anterior to the eyes. After removing the connective tissue and the muscles with forceps, use the end of a sterile cotton swab as a regime to identify the bregma and the coronal sutures.
After placing the armed retractor, place a 25-gauge spinal needle in the stereotaxic frame exactly on the bregma and note the position. Next, remove the needle from the bregma. Move the frame anteriorly by 6.5 millimeters, then replace the needle in the midline to mark the side of drilling.
Drill until the dura mater is identified below the bone. Using straight forceps, gently remove the bone fragments, then fill the cavity with bone wax. Next, 3 to 4 millimeters lateral to the right of the bregma and just anterior to the coronal suture for the laser Doppler, drill another hole taking care not to penetrate the dura mater.
Look for the vessels where the laser Doppler can measure the blood flow. Place the laser Doppler and check the values. A minimum value of 100 flux units is required. If the values remain acceptable after removing the microscope, add 1 drop of glue to fix the probe. Recheck to confirm whether the value is above 80 flux units.
To induce subarachnoid hemorrhage, insert the needle gently through the skull in the midline between the hemispheres until resistance is felt due to the base of the skull. Retract the needle by 1 millimeter to ensure correct placement just anteriorly to the optic chiasm. To ensure the most homogeneous result when injecting the blood, turn the needle clockwise by 90 degrees such that the needle tip points to the right. Then, remove the stiletto.
After a 15-minute equilibration, adjust the level of anesthesia to obtain a mean arterial blood pressure in the range 80 to 100 millimeters of mercury. Then, perform a blood gas analysis to confirm that the pH, partial pressure of carbon dioxide and partial pressure of oxygen are within physiological parameters.
Next, using a 1-milliliter syringe with a blunt 23-gauge needle, withdraw 500 microliters of blood from the tail catheter. To avoid injection of air, fill the dead space of the spinal needle chamber with blood. After adjusting the volume of the blood in the syringe to 300 microliters, connect the syringe to the spinal needle. Then, grasping firmly around the syringe, inject the blood manually to surpass mean arterial pressure. Observe a steep rise in the intracerebral pressure and a concurrent steep fall in the cerebral blood flow.