Name: continuous iv infusion is the choice treatment route for arginine-vasopressin receptor blocker conivaptan in mice to study stroke-evoked brain edema
Uploaded: 2016-09-01T14:00:00.000+00:00
Duration: 8 min 44 s
Description: Watch this Scientific Journal Video about Continuous IV Infusion is the Choice Treatment Route for Arginine-vasopressin Receptor Blocker Conivaptan in Mice to Study Stroke-evoked Brain Edema at JoVE.c

我们感谢瑞典医疗中心提供资金和设施。我们也感谢克雷格医院的大量使用的实验室空间。

实验是按照美国国立卫生研究院的照顾和使用动物研究的指导方针进行，由瑞典医学中心的动物护理和使用委员会批准。所有的程序都用适当的无菌技术进行的。这项研究使用的实验动物均为男性，3个月大，体重野生型C57小鼠25日至27克 1. 在体内诱导中风<ol><li>预涂层与树脂牙灯丝此前的手术。 <ol><li>剪切从7-0尼龙线12毫米长片丝。混合2份该树脂与硬化剂的1份，然后立即浸长丝的前端到混合物，以覆盖整个长度的约1/3。快速清除并验证它覆盖的牙科树脂的表面光滑的外套。 </li><li>风干灯丝在使用前2小时。 </li></ol></li><li>将鼠标进入Ñ​​麻醉诱导室（2L体积），并设置在2升/分钟进入腔室1.5％异氟烷在25％的富氧室内空气的流动。 </li><li>允许小鼠留在感应室中直至完全麻醉，8由缺乏响应于尾捏确定。 </li><li>将鼠标走上手术台用加热垫，并设定1％的异氟醚浓度维持通过鼻锥自然娩出麻醉。 </li><li>夹上的前部和颈部的背毛，并用酒精喷颈部。在两只眼睛应用兽医眼药膏。 </li><li>连续监测体温，用在手术过程中连接到数字显示直肠温度探头。 </li><li>鼠标放置在仰卧位，应用碘伏10％溶液擦洗消毒，用无菌悬垂盖，然后使沿颈部正中切口，大小10手术刀刀片。 </li> &lt;利&gt;暂时结扎左颈总动脉（CCA）以6.0缝合丝线。 </li><li>放置在左侧颈外动脉（ECA）的另一个领带。 </li><li>放置在颈内动脉（ICA）微血管剪辑。 </li><li>削减与微血管剪刀颈外动脉一小的孔。插入来自涂上树脂牙将在ECA开幕7-0尼龙线做灯丝，并cephalically推进它，同时消除夹，直到感到有阻力（从CCA分叉处约5-7毫米）。 </li><li>放置一个临时粘结到所述内颈动脉，以确保灯丝中的位置。参考下图， 图1A。收用丝线缝合皮肤，并渗透在颈部的前部的伤口用0.2ml 0.5％布比卡因，如在步骤2.6。 </li><li>让鼠标从麻醉中苏醒并恢复意识在恢复室中60分钟。不要离开无人看管的动物，直到它已经恢复足够的意识，以保持胸骨靠着。 </li><li>测试鼠标的神经功能缺损评分（NDS）如下:0 =运动功能正常，躯干和尾板对侧前肢1 =前屈，2 =盘旋至对侧，但在休息，3正常坐姿=偏向对侧侧静止的，4 =无自发运动活性。4,9 <ol><li>对于NDS测试，尾巴和地点在一个平面上进行观察（2-3分钟）抬起鼠标。另外，观察在恢复室​​老鼠。排除表现出比NDS 2从实验降低因MCA闭塞不足小鼠。 注意:激光多普勒血流仪技术可以被用来确认MCA闭塞9-11。 </li></ol></li><li>重新麻醉如上所述动物后在感应腔室恢复60分钟，再消毒用10％聚维酮碘溶液擦洗先前切开皮肤，并重新打开在手术伤口通过切割和去除丝线缝合颈部的前面。 </li><li>从ECA取出长丝执行以下步骤: <ol><li>广场上的ICA微血管剪辑。解开对ICA的缝线拉出长丝的同时抱夹打开。 </li><li>关闭剪辑和近端放置领带到非洲经委会晋级。 </li><li>除去夹子和从CCA除去结扎以恢复血流到大脑。参考下图， 图1B。 </li></ol></li></ol> 2.安装静脉导管进入颈静脉连续48小时处理注:步骤1.16.3没有鼠标醒后立即进行安装导管。 <ol><li>使用柔性管的两种不同的尺寸:插入所述内部管道（外径0.94毫米;​​以包含药物被注入）到外管（外径3.18毫米;对于内管的保护）。 </li><li> P花边鼠标在俯卧位，适用10％聚维酮碘溶液擦洗消毒，并在脖子上的，大小10手术刀刀片后面有1厘米的切口。转小鼠成仰卧位，重新消毒用10％聚维酮碘溶液擦洗颈部的前部，并重新打开在颈部的前部的外科手术伤口。隧道从背面到颈部的前所作的切口在皮肤下的管道，并从打开的伤口1厘米exteriorize它。 </li><li>从颈部的前部横向约1cm距离中线的左边除去皮下脂肪组织。找到左侧颈内静脉，将两条领带沿静脉相距5mm，并且略微拉长静脉。就微血管剪刀两个关系之间的颈静脉一个小孔;插入内管5 mm深尾端尖（朝心脏）。 </li><li>固定管与两个联系的颈静脉。关闭皮肤在颈部的前端与3-0丝线缝合。参考下图， 图1C。 </li><li>固定油管的颈部与缝合背面的皮肤，并通过一个旋转连接管连接到一个微注射静脉泵。旋转允许在笼子里面有完全访问食物和水的小鼠自由流动。 </li><li>渗透前面的皮肤切口和颈部用0.2ml 0.5％布比卡因防止手术后疼痛的背面。 注:根据机构特定动物的协议，动物可能与阿片类药物，NSAIDs类药物或抗生素前和手术后进行治疗。然而，提及的治疗可能会改变缺血性脑卒中结局结果 （见讨论部分 ）11-14 </li><li>让动物在回收室唤醒，然后安置动物在一个单独的笼子，直到实验结束点。不要让动物无人看管，直到它已经恢复了足够的意识，以保持胸骨靠着。 </li><li>以1.5ml / kg /小时，4设置为连续48小时静脉治疗考尼伐坦的（0.14毫克/毫升，在5％右旋糖）或车辆输注速度和开始输注。 注意:四输注考尼伐坦的总量应该大约为0.2毫克/小鼠/天，1.44毫升总体积。 注意:为0.2毫克总量在1.44毫升5％右旋糖，每天两次执行考尼伐坦腹腔（IP）注射。实现IP注射如前所述。15通过抓住主体和尾部的后表面的皮肤抑制的小鼠。注入考尼伐坦或车辆进入使用26 G / 1 /寸毫针腹部的左下腹。15 </li><li>观察动物每天两次的实验（48小时）的整个持续时间。让动物充分获得食物和水。如果动物表现出痛苦或呼吸困难的迹象，请教兽医。如果安乐死是必要的严重DIST动物RESS参考以下步骤3.1，并排除在研究动物。 </li></ol>在终点中风诱发脑水肿3.评估<ol><li>通过用5％异氟醚的过麻醉牺牲鼠标。 </li><li>使用手术刀，使过沿中线颅骨切开皮肤，并缩回皮以暴露在大脑中的整个颅骨。在枕骨大孔开始，切开颅骨沿两侧大脑的周边横向，然后小心地抬起而不触及它下面的大脑。切断从脊柱脑和附着在颅骨的基础上的神经。 </li><li>取出大脑，它从嗅球和小脑分开，并解剖沿纵裂大脑进入缺血和非缺血大脑半球，如前所述。9 </li><li>通过如前面描述的4,9比较湿到干比（WDR）评估脑水肿。权衡组织之前和在烘箱中在100℃下干燥3天。计算脑含水量（BWC）的％H 2 O =（1-干重/湿重）×100％。 </li></ol>

<ol>
	<li>Gueniau, C., Oberlander, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+kappa+opioid+agonist+niravoline+decreases+brain+edema+in+the+mouse+middle+cerebral+artery+occlusion+model+of+stroke.">The kappa opioid agonist niravoline decreases brain edema in the mouse middle cerebral artery occlusion model of stroke.</a> J Pharmacol Exp Ther. 282, 1-6 (1997).</li><li>Krafft, P. R., 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=Etiology+of+stroke+and+choice+of+models.">Etiology of stroke and choice of models.</a> Int J Stroke. 7, 398-406 (2012).</li><li>Vakili, A., Kataoka, H., Plesnila, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+arginine+vasopressin+V1+and+V2+receptors+for+brain+damage+after+transient+focal+cerebral+ischemia.">Role of arginine vasopressin V1 and V2 receptors for brain damage after transient focal cerebral ischemia.</a> J Cereb Blood Flow Metab. 25, 1012-1019 (2005).</li><li>Zeynalov, E., Jones, S. M., Seo, J. W., Snell, L. D., Elliott, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arginine-Vasopressin+Receptor+Blocker+Conivaptan+Reduces+Brain+Edema+and+Blood-Brain+Barrier+Disruption+after+Experimental+Stroke+in+Mice.">Arginine-Vasopressin Receptor Blocker Conivaptan Reduces Brain Edema and Blood-Brain Barrier Disruption after Experimental Stroke in Mice.</a> PloS one. 10, e0136121 (2015).</li><li>Med Lett Drugs Ther.. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conivaptan+(Vaprisol)+for+hyponatremia.">Conivaptan (Vaprisol) for hyponatremia.</a> The Medical letter on drugs and therapeutics. 48, 51-52 (2006).</li><li>Zhao, X. Y., 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=Effect+of+arginine+vasopressin+on+the+cortex+edema+in+the+ischemic+stroke+of+Mongolian+gerbils.">Effect of arginine vasopressin on the cortex edema in the ischemic stroke of Mongolian gerbils.</a> Neuropeptides. 51, 55-62 (2015).</li><li>Manaenko, A., Chen, H., Kammer, J., Zhang, J. H., Tang, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+Evans+Blue+injection+routes:+Intravenous+versus+intraperitoneal,+for+measurement+of+blood-brain+barrier+in+a+mice+hemorrhage+model.">Comparison Evans Blue injection routes: Intravenous versus intraperitoneal, for measurement of blood-brain barrier in a mice hemorrhage model.</a> J Neurosci Methods. 195, 206-210 (2011).</li><li>Adams, S., Pacharinsak, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+anesthesia+and+analgesia.">Mouse anesthesia and analgesia.</a> Curr Protoc Mouse Biol. 5, 51-63 (2015).</li><li>Zeynalov, E., 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=The+perivascular+pool+of+aquaporin-4+mediates+the+effect+of+osmotherapy+in+postischemic+cerebral+edema.">The perivascular pool of aquaporin-4 mediates the effect of osmotherapy in postischemic cerebral edema.</a> Crit Care Med. 36, 2634-2640 (2008).</li><li>Zeynalov, E., Dore, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low+doses+of+carbon+monoxide+protect+against+experimental+focal+brain+ischemia.">Low doses of carbon monoxide protect against experimental focal brain ischemia.</a> Neurotox Res. 15, 133-137 (2009).</li><li>Zeynalov, E., Nemoto, M., Hurn, P. D., Koehler, R. C., Bhardwaj, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuroprotective+effect+of+selective+kappa+opioid+receptor+agonist+is+gender+specific+and+linked+to+reduced+neuronal+nitric+oxide.">Neuroprotective effect of selective kappa opioid receptor agonist is gender specific and linked to reduced neuronal nitric oxide.</a> J Cereb Blood Flow Metab. 26, 414-420 (2006).</li><li>Ma, M. C., Qian, H., Ghassemi, F., Zhao, P., Xia, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oxygen-sensitive+{delta}-opioid+receptor-regulated+survival+and+death+signals:+novel+insights+into+neuronal+preconditioning+and+protection.">Oxygen-sensitive {delta}-opioid receptor-regulated survival and death signals: novel insights into neuronal preconditioning and protection.</a> J Biol Chem. 280, 16208-16218 (2005).</li><li>Ahmad, M., Zhang, Y., Liu, H., Rose, M. E., Graham, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prolonged+opportunity+for+neuroprotection+in+experimental+stroke+with+selective+blockade+of+cyclooxygenase-2+activity.">Prolonged opportunity for neuroprotection in experimental stroke with selective blockade of cyclooxygenase-2 activity.</a> Brain Res. 1279, 168-173 (2009).</li><li>Meisel, C., 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=Preventive+antibacterial+treatment+improves+the+general+medical+and+neurological+outcome+in+a+mouse+model+of+stroke.">Preventive antibacterial treatment improves the general medical and neurological outcome in a mouse model of stroke.</a> Stroke. 35, 2-6 (2004).</li><li>Miner, N. A., Koehler, J., Greenaway, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraperitoneal+injection+of+mice.">Intraperitoneal injection of mice.</a> Appl Microbiol. 17, 250-251 (1969).</li><li>Adis International Limited. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conivaptan:+YM+087.">Conivaptan: YM 087.</a> Drugs in R&amp;D. 5, 94-97 (2004).</li><li>Murphy, T., Dhar, R., Diringer, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conivaptan+bolus+dosing+for+the+correction+of+hyponatremia+in+the+neurointensive+care+unit.">Conivaptan bolus dosing for the correction of hyponatremia in the neurointensive care unit.</a> Neurocrit Care. 11, 14-19 (2009).</li><li>Liu, X., Nakayama, S., Amiry-Moghaddam, M., Ottersen, O. P., Bhardwaj, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arginine-vasopressin+V1+but+not+V2+receptor+antagonism+modulates+infarct+volume,+brain+water+content,+and+aquaporin-4+expression+following+experimental+stroke.">Arginine-vasopressin V1 but not V2 receptor antagonism modulates infarct volume, brain water content, and aquaporin-4 expression following experimental stroke.</a> Neurocrit Care. 12, 124-131 (2010).</li><li>Wallace, B. K., Jelks, K. A., O'Donnell, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ischemia-induced+stimulation+of+cerebral+microvascular+endothelial+cell+Na-K-Cl+cotransport+involves+p38+and+JNK+MAP+kinases.">Ischemia-induced stimulation of cerebral microvascular endothelial cell Na-K-Cl cotransport involves p38 and JNK MAP kinases.</a> Am J Physiol Cell Physiol. 302, C505-C517 (2012).</li><li>O'Donnell, M. E., 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=Intravenous+HOE-642+reduces+brain+edema+and+Na+uptake+in+the+rat+permanent+middle+cerebral+artery+occlusion+model+of+stroke:+evidence+for+participation+of+the+blood-brain+barrier+Na/H+exchanger.">Intravenous HOE-642 reduces brain edema and Na uptake in the rat permanent middle cerebral artery occlusion model of stroke: evidence for participation of the blood-brain barrier Na/H exchanger.</a> J Cereb Blood Flow Metab. 33, 225-234 (2013).</li><li>Walcott, B. P., Kahle, K. T., Simard, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+treatment+targets+for+cerebral+edema.">Novel treatment targets for cerebral edema.</a> Neurotherapeutics. 9, 65-72 (2012).</li><li>Shen, 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=Inhibition+of+G+protein-coupled+receptor+81+(GPR81)+protects+against+ischemic+brain+injury.">Inhibition of G protein-coupled receptor 81 (GPR81) protects against ischemic brain injury.</a> CNS Neurosci Ther. 21, 271-279 (2015).</li></ol>

作者什么都没有透露。

这项研究有临床前研究中风重要的价值。这项研究表明，小鼠实验中风后考尼伐坦（0.2毫克/天）的持续静脉注射，有效降低治疗后的48小时脑水肿。 IP注射相同剂量对脑水肿考尼伐坦的效果也进行了研究。考尼伐坦治疗由双方IV和IP路由产生aquaresis由所示小鼠:1）血浆渗透压略高于生理水平的提高;和2）在尿渗透压降低由于在肾水再吸收的堵塞。因此，基于对治疗结果，我们报告中指出，四交付方?...

Stroke continues to be an enormous burden for patients and clinicians. Animal stroke models have been used in the laboratory setting for nearly two decades.1 Nevertheless, experimental treatments that work in animals often fail in humans.2 This discrepancy in treatment outcomes may be due to various factors, such as administration routes for drugs used in animal research, drug metabolism and elimination rate, and many other aspects. One of the major complications of stroke, brain edema, is a focus of current research in neuroscience. Mechanisms of brain edema formation involve disturbances in water and electrolyte balance triggered by the arginine-vasopressin (AVP) response to ischemic brain injury.3 Therefore, AVP and its receptors (V1a and V2) are a major focus of research studies of post-ischemic brain edema formation.3
We have developed a methodology to study the effects of mixed arginine-vasopressin (V1a and V2) receptor blocker conivaptan on post-ischemic brain edema in mice.4 Molecular targets of conivaptan5 make the drug a suitable candidate for exploration of its properties in alleviation of brain edema. Furthermore, conivaptan needs to be studied in the context of pathophysiological events of stroke.6 In designing this study, we considered comparing treatment outcomes using two different routes of administration for conivaptan: intravenous (IV)4 and intraperitoneal (IP).7 Effects of the treatments on stroke-induced brain edema were evaluated. Here detailed protocols are described for surgical induction of experimental stroke by middle cerebral artery occlusion (MCAO), and followed by continuous conivaptan treatment using the installation of a jugular IV catheter. After induction of stroke, animals were randomly allocated into the following groups: vehicle or conivaptan (0.2 mg/day) treated IV or IP.

<table><tbody><tr><td>Heated Pad</td><td>K&amp;amp;H Manufacturing Inc</td><td>1060</td><td></td></tr><tr><td>Temperature Monitor with Rectal Probe</td><td>Physitemp</td><td>7029</td><td></td></tr><tr><td>Silk Suture Spool, 6-0</td><td>Surgical Specialties Corporation</td><td>SP114</td><td></td></tr><tr><td>Silk Suture on a Needle, 3-0</td><td>Ethicon</td><td>1684G</td><td></td></tr><tr><td>Nylon Suture, 7-0</td><td>Ethicon</td><td>1696G</td><td></td></tr><tr><td>Dental Resin Polysiloxane with Hardener</td><td>Heraeus Kulzer</td><td>65817930</td><td></td></tr><tr><td>Microinfusion IV Pump</td><td>Kent Scietific</td><td>GT0897</td><td></td></tr><tr><td>Swivel 22GA</td><td>Instech</td><td>375/22PS</td><td></td></tr><tr><td>Laboratory Tubing, 0.94 mm x 0.51 mm</td><td>Dow Corning</td><td>508-002</td><td></td></tr><tr><td>Laboratory Tubing, 3.18 mm x 1.98 mm</td><td>Dow Corning</td><td>508-009</td><td></td></tr></tbody></table>

continuous iv infusion is the choice treatment route for arginine-vasopressin receptor blocker conivaptan in mice to study stroke-evoked brain edema

中风是在世界发病和死亡的主要原因之一。中风是脑水肿等病理生理事件复杂化。间在中风诱发的脑水肿的发展和演变的最重要的球员是激素精氨酸加压素及其受体，V1a和V2。最近，V1a和V2受体阻滞剂考尼伐坦受到关注作为潜在的药物，以减少中风后的脑水肿。然而，这在中风研究涉及考尼伐坦应用动物模型需要根据给药可行途径进行修改。在这里，48小时连续静脉注射（IV）的结果与小鼠实验中风后腹腔内（IP）考尼伐坦治疗相比。我们开发了在大脑中动脉闭塞与导管安装组合为IV治疗考尼伐坦（0.2毫克）或车辆的颈静脉的协议。动物的不同队列，用0.2治疗考尼伐坦或车辆的IP毫克，每天丸。实验中风诱发脑水肿连续IV和IP处理后的小鼠进行了评估。的结果比较，可知考尼伐坦的连续IV施用减轻小鼠局部缺血后的脑水肿，与考尼伐坦的IP施用。我们得出结论，我们的模型，可用于在中风和脑水肿的上下文考尼伐坦应用今后的研究。

动物的体温是在生理范围内，并且在整个冲程感应的手术过程稳定。两个小鼠表现出NDS低于2 MCAO从该研究中排除之后。 在缺血小鼠48小时生产同侧半球梗死体积。的TTC染色切片的评价表明，半球的约50％是由缺血（ 图1D）后梗塞的影响，如通过Zeynalov，E。和多尔，S。，2009年10脑缺血性中风的发病引起严重?...

Watch this Scientific Journal Video about Continuous IV Infusion is the Choice Treatment Route for Arginine-vasopressin Receptor Blocker Conivaptan in Mice to Study Stroke-evoked Brain Edema at JoVE.c

Continuous IV Infusion is the Choice Treatment Route for Arginine-vasopressin Receptor Blocker Conivaptan in Mice to Study Stroke-evoked Brain Edema

Our studies have revealed that the beneficial effects of conivaptan are dependent on the method of delivery after experimental stroke in mice. We have developed a research protocol for delivery of the receptor blocker via IV catheter on stroke-evoked brain edema formation in mice.

持续静脉注射是精氨酸加压素受体拮抗剂考尼伐坦的选择治疗途径小鼠来研究中风诱发脑水肿

Stroke is one of the major causes of morbidity and mortality in the world. Stroke is complicated by brain edema and other pathophysiological events. Among ...

continuous-iv-infusion-is-choice-treatment-route-for-arginine

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

8.8K 次观看.  Swedish Medical Center. Our studies have revealed that the beneficial effects of conivaptan are dependent on the method of delivery after experimental stroke in mice. We have developed a research protocol for delivery of the receptor blocker via IV catheter on stroke-evoked brain edema formation in mice. 

视频: Continuous IV Infusion is the Choice Treatment Route for Arginine-vasopressin Receptor Blocker Conivaptan in Mice to Study Stroke-evoked Brain Edema

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) ...

科研

医学

Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. Intra-tracheal inoculation of a known amount of bacteria has been successfully used to reproduce the pathogenesis of pneumonia which then develops into sepsis. Monitoring hemodynamic parameters and providing standard clinical treatment including infusion of antibiotics, fluids and drugs to maintain blood pressure is critical to simulate routine supportive care in this model but to do so requires both arterial and venous vascular access. The video details the surgical technique for implanting carotid artery and common jugular vein catheters in an anesthetized rat. Following a 72 hr recovery period, the animals will be re-anesthetized and connected to a tether and swivel setup attached to the rodent housing which connects the implanted catheters to the hemodynamic monitoring system. This setup allows free movement of the rat during the study while continuously monitoring pressures, infusing fluids and drugs (antibiotics, vasopressors) and performing blood sampling.

Vascular accesses to measure hemodynamics, provide fluids and perform blood sampling are important to any small animal model study. We present a technique for implanting catheters into the carotid artery and the common jugular vein in an anesthetized rat for connecting to a system to perform monitoring, infusions and sampling.

颈动脉和颈静脉导管在有意识大鼠模型进行血流动力学措施，输液和血液采样

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. ...

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 ...

An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke

Ischemic stroke leads to vasogenic cerebral edema and subsequent primary brain injury, which is mediated through destruction of the blood-brain barrier (BBB). Rats with induced ischemic stroke were established and used as in vivo models to investigate the functional integrity of the BBB. Spectrophotometric detection of Evans blue (EB) in the brain samples with ischemic injury could provide reliable justification for the research and development of novel therapeutic modalities. This method generates reproducible results, and is applicable in any laboratory without a need for special equipment. Here, we present a visualized and technical guideline on the detection of the extravasation of EB following induction of ischemic stroke in rats.

An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke

The overall goal of this procedure is to provide a highly reproducible technique for in vivo assessment of the blood-brain barrier disruption in rat models of ischemic stroke.

一种体内评价大鼠缺血性中风模型中血脑屏障的破坏

Ischemic stroke leads to vasogenic cerebral edema and subsequent primary brain injury, which is mediated through destruction of the blood-brain barrier ...

神经科学

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 ...

Combined Infusion and Stimulation with Fast-Scan Cyclic Voltammetry (CIS-FSCV) to Assess Ventral Tegmental Area Receptor Regulation of Phasic Dopamine

Phasic dopamine (DA) release from the ventral tegmental area (VTA) to the nucleus accumbens plays a pivotal role in reward processing and reinforcement learning. Understanding how the diverse neuronal inputs into the VTA control phasic DA release can provide a better picture of the circuitry that controls reward processing and reinforcement learning. Here, we describe a method that combines intra-VTA cannula infusions of pharmacological agonists and antagonists with stimulation-evoked phasic DA release (combined infusion and stimulation, or CIS) as measured by in vivo fast-scan cyclic voltammetry (FSCV). Using CIS-FSCV in anesthetized rats, a phasic DA response can be evoked by electrically stimulating the VTA with a bipolar electrode fitted with a cannula while recording in the nucleus accumbens core. Pharmacological agonists or antagonists can be infused directly at the stimulation site to investigate specific VTA receptors&#39; roles in driving phasic DA release. A major benefit of CIS-FSCV is that VTA receptor function can be studied in vivo, building on in vitro studies.

Combined Infusion and Stimulation with Fast-Scan Cyclic Voltammetry CIS-FSCV to Assess Ventral Tegmental Area Receptor Regulation of Phasic Dopamine

The goal of this protocol is to directly manipulate ventral tegmental area receptors to study their contribution to subsecond dopamine release.

联合输注和刺激与快速扫描循环伏安法（CIS-FSCV）评估腹侧被盖区域受体对相位多巴胺的调节

Phasic dopamine (DA) release from the ventral tegmental area (VTA) to the nucleus accumbens plays a pivotal role in reward processing and reinforcement ...

Unraveling the Role of Discrete Areas of the Rat Brain in the Regulation of Ovulation through Reversible Inactivation by Tetrodotoxin Microinjections

Many experimental approaches have been used for studying the role of the brain in the regulation of ovulation. Examples include the lesion and deafferentation of neuronal groups, which are both invasive methods that permanently impair the integrity of the target area. These methods are accompanied by collateral effects that can affect the analysis of acute and temporal regulatory mechanisms. The stereotaxic implantation of guide cannulas aimed at specific brain regions, followed by a recovery period, allows researchers to microinject different drugs after the disappearance of the undesired effects of the surgery. Tetrodotoxin has been used to determine the roles of several brain areas in diverse physiological processes because it transiently inhibits the sodium-dependent action potentials, thus blocking all neural activity in the target region. This protocol combines this method with strategies for the assessment of the estrous cycle and ovulation to reveal the role of discrete brain regions in the regulation of ovulation at particular times of any given stage of the estrous cycle. Awake and unrestrained rats (Rattus norvegicus) were used to avoid the blocking effects that anesthetics and stress hormones exert on ovulation. This protocol can be easily adapted to other species, brain targets and pharmacological agents to study different physiological processes. Future improvements to this method include the design of a microinjection system using glass capillaries of small diameter instead of guide cannulas. This will reduce the amount of tissue damaged during the implantation and decrease the spread of the infused drugs outside the target area.

This protocol describes the construction of a low-cost microinjection system, its stereotaxic implantation into deep-brain structures and the procedure for timed microinjections of tetrodotoxin in awake and unrestrained rats. The goal is to reveal the participation of hypothalamic structures in the regulation of ovulation by inhibiting their neural activity.

通过特洛多毒素微注射可逆灭活，揭开鼠脑离散区在排卵调节中的作用

Many experimental approaches have been used for studying the role of the brain in the regulation of ovulation. Examples include the lesion and ...

Intracranial Pressure Monitoring In Nontraumatic Intraventricular Hemorrhage Rodent Model

Survivors of intraventricular hemorrhage are often left with significant long-term memory impairment; thus, research utilizing intraventricular hemorrhage animal models is essential. In this study, we sought out ways to measure intracranial pressure, mean arterial pressure, and cerebral perfusion pressure during nontraumatic intraventricular hemorrhage in rats. The experimental design included three Sprague Dawley groups: sham, standard 200 &#181;l intraventricular hemorrhage, and vehicle control groups. By introducing an intraparenchymal fiberoptic pressure sensor, precise intracranial pressure measurements were obtained in all groups. Cerebral perfusion pressures were calculated with the knowledge of intracranial pressure and mean arterial pressure values. As expected, the intraventricular hemorrhage and vehicle control groups both experienced a rise in the intracranial pressure and subsequent decline in cerebral perfusion pressure during intraventricular injection of autologous blood and artificial cerebrospinal fluid, respectively. The addition of an intraparenchymal fiberoptic pressure sensor is beneficial in monitoring precise intracranial pressure changes.

Monitoring intracranial pressure in rodent&#160;models of nontraumatic intraventricular hemorrhage is not common in the current literature. Herein, we demonstrate a technique for measuring intracranial pressure, mean arterial pressure, and cerebral perfusion pressure during intraventricular hemorrhage in a rat&#160;animal model.

非创伤性脑室内出血啮齿动物模型中的颅内压监测

Survivors of intraventricular hemorrhage are often left with significant long-term memory impairment; thus, research utilizing intraventricular hemorrhage ...

Intraventricular Drug Delivery and Sampling for Pharmacokinetics and Pharmacodynamics Study

Although the blood-brain barrier (BBB) protects the brain from foreign entities, it also prevents some therapeutics from crossing into the central nervous system (CNS) to ameliorate diseases or infections. Drugs are administered directly into the CNS in animals and humans to circumvent the BBB. The present protocol describes a unique way of treating brain infections through intraventricular delivery of antibiotics, i.e., polymyxins, the last-line antibiotics to treat multi-drug resistant Gram-negative bacteria. A straightforward stereotaxic surgery protocol was developed to implant a guide cannula reaching into the lateral ventricle in rats. After a recovery period of 24 h, rats can be injected consciously and repeatedly through a cannula that is fitted to the guide. Injections can be delivered manually as a bolus or infusion using a microinjection pump to obtain a slow and controlled flow rate. The intraventricular injection was successfully confirmed with Evans Blue dye. Cerebrospinal fluid (CSF) can be drained, and the brain and other organs can be collected. This approach is highly amenable for studies involving drug delivery to the CNS and subsequent assessment of pharmacokinetic and pharmacodynamic activity.

<meta charset="utf8"></head><body>用于药代动力学和药效学研究的脑室内药物输送和取样

Delivery of therapeutics directly into the central nervous system is one way of circumventing the blood-brain barrier. The present protocol demonstrates intracerebroventricular injection for subsequent collection of cerebrospinal fluid and bodily organs. This facilitates the investigation of drug pharmacokinetics and pharmacodynamics in animal models for developing new treatments.

用于药代动力学和药效学研究的脑室内药物输送和取样

Although the blood-brain barrier (BBB) protects the brain from foreign entities, it also prevents some therapeutics from crossing into the central nervous ...

持续静脉注射是精氨酸加压素受体拮抗剂考尼伐坦的选择治疗途径小鼠来研究中风诱发脑水肿

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此视频中的章节

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

颈动脉和颈静脉导管在有意识大鼠模型进行血流动力学措施，输液和血液采样

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

一种体内评价大鼠缺血性中风模型中血脑屏障的破坏

小鼠蛛网膜下腔出血模型脑血管痉挛量化的容积法

联合输注和刺激与快速扫描循环伏安法（CIS-FSCV）评估腹侧被盖区域受体对相位多巴胺的调节

通过特洛多毒素微注射可逆灭活，揭开鼠脑离散区在排卵调节中的作用

非创伤性脑室内出血啮齿动物模型中的颅内压监测

用于药代动力学和药效学研究的脑室内药物输送和取样

用于药代动力学和药效学研究的脑室内药物输送和取样