这项工作是由美国国立卫生研究院[格兰特没有支持。 R01NS069726]和美国心脏协会的资助以资助格兰特[格兰特没有。 13GRNT17020004]到SD。

注:小鼠（C57BL / 6J，男）岁 10 - 19周后在该研究中使用。所有的程序均按照美国国立卫生研究院指南实验动物的护理和使用执行和批准了密苏里州机构动物护理和使用委员会的大学（IACUC）。 1.手术前<ol><li>手术前的高压釜一天消毒所有的手术器械。裹在121℃下的仪器和高压灭菌15磅30分钟，接着干燥（121℃，15磅，30/30循环）的30分钟。将在清洁无菌的环境仪器，直到进一步使用。 </li><li>准备新鲜的玫瑰红（RB）溶液（20毫克/毫升无菌生理盐水）在手术前的每一次。以完全溶解的RB，涡流管再接着超声处理5分钟，在50/60赫兹为19 W.裹管在铝箔和保护其不受光输出功率，直至河畔期间进一步使用盖瑞。 </li><li>制备氯胺酮/甲苯​​噻嗪的混合物在无菌盐水中。添加125微升甲苯噻嗪（原液浓度:20毫克/毫升）和325微升ketmaine的（原液浓度:100毫克/毫升）和550微升无菌生理盐水，使1毫升麻醉剂混合物的最终体积。 </li><li>预暖恒温加热垫。 </li><li>预温的金属卤化物灯（光源FN1落射荧光显微镜）处理30分钟，以稳定的灯功率。 </li><li>调整照明区域的大小的直径的使用10X物镜和一个分划板通过调整视场光阑的直立FN1落射荧光显微镜1毫米。 </li></ol> 2.手术过程<ol><li>麻醉小鼠用氯胺酮（130毫克/千克B.重量）和甲苯噻嗪的剂量（10毫克/千克B.重量）。基于鸡尾酒在1.3，每克鼠标重量B. 4微升。是必要的。消毒注射部位用酒精棉球，并通过帧内施用麻醉剂-peritoneal（IP）的路由。小心不要将麻醉药注入到血管或肌肉，因为这会耽误动物的诱导和恢复。 </li><li>应用人工泪液软膏鼠标的双眼，以防止干燥，然后将动物在加热垫，以防止体温过低。 </li><li>动物的制备<ol><li>通过脚趾捏响应检查动物进行适当的手术麻醉的水平。 </li><li>一旦动物已经达到麻醉手术水平，使用的是电动毛发修剪器夹上围绕动物的中线的背侧表面上的毛发。擦洗手术部位用70％乙醇，随后通过优碘溶液三次。遮盖的部位用无菌手术悬垂至下一个步骤。 </li></ol></li><li>手术瘦骨暴露脊髓<ol><li>鼠标放置在通过外科手术平台（ 图1A）上的恒温加热垫俯卧姿势。正确保护鼠标使用吻钳姿势维持一个细长颈区（ 图1A，B）。 </li><li>做一个切口（约1厘米长）使用沿着从胸椎到T9 T12延长背中线手术剪。走到一边的皮肤暴露手术区。 </li><li>使用手术刀，仔细清除肌肉，露出背刺在T9 - T12椎体。停在每一步运用所用无菌棉签轻压止血。单独T10 - T12椎骨从周围的肌肉和使用椎夹具来稳定和防止任何移动它们固定（ 图1A，B）。 </li><li>使用高速钻骨抛光钻头，小心翼翼地轻轻薄T10 T11或脊椎的背部表面可视化的SC（ 图1C）的背水面后脊柱静脉和其他小型船只。 </li><li>为了防止热损伤是由于减薄过程期间产生的热，应用生理盐水的温柔络绎不绝随着不断的吸取出碎片。 </li><li>使用解剖刀小心平滑骨表面，直到主容器是清晰可见。注意不要在此过程中损伤脊髓。 </li><li>一旦血管被可视化，管理的RB在通过眶后窦路线使用胰岛素注射器的剂量为30毫克/公斤（体重）。 </li><li>使用激光多普勒流量计3分钟后以下的RB注射测量血流量，如果必要（ 图2A，B）。在保持整个过程的无菌。 </li></ol></li></ol> 3.感应PT的<ol><li>将动物在XY位置可调级过一个实验室千斤顶可调节高度。调整鼠标的位置，从而T11脊髓的暴露区域是直属FN1落射荧光显微镜（ 图3A）的10倍的目标。 </li><li>设置里的力量向右源在12％，并与在减薄脊髓的中间直径为0.75mm照射T11区域（注:此区域包括后脊柱静脉和其他毛细血管）与绿色光（波长540 - 580纳米，这是通过在显微镜过滤立方体），通过10倍的目标2分钟完成。拍摄图像的开头和照射端（ 图3B，C）和在此点记录实验的时间。 </li><li>再次测量血流量为10分钟，如果必要通过将激光多普勒探针的相同位置的脊髓上面在2.4.8（ 图2A，B）。 </li><li>照射检查是否有出血及后若没有发现进行缝合动物。缝合浅筋膜连同使用可吸收的缝合线或4-0大小丝线对脊髓的任一侧上的肌肉。小心不要破坏暴露SC。缝合用4-0丝线缝合皮肤。应用聚维酮碘或碘到缝合后皮肤的边缘。 </li></ol> 4.手术后护理<ol><li>缝合后，放置在加热垫进行恢复的动物。恢复后通过观察两个后肢肢体运动检查动物的神经功能缺损的迹象。不要让动物无人看管，直到它已经恢复了意识足以维持胸骨斜卧。 </li><li>将动物转移到饲养笼。不返回已经历手术给其他动物的公司，直到完全恢复的动物。 </li><li>检查动物定期。在情况严重的神经功能障碍，提供适当的照顾就像疏散膀胱，止痛药的管理（丁丙诺啡，0.05 - 0.1毫克/千克）。检查脱水和管理生理盐水皮下在严重的情况。一般，丁丙诺啡（0.1毫克/千克）将缝合，以减轻疼痛，在外科手术部位后给药。 </li><li>如果动物是不能立即手术后安乐死，我们会放在笼地板含水量高的饮食，让动物能轻易达到的食物。 </li></ol> 5.穿心灌注，尼氏染色和免疫染色<ol><li> Transcardially灌注动物如前面所述17-20。 <ol><li>麻醉动物在协议中前面描述和用磷酸盐缓冲盐水（PBS，pH 7.4）中，随后冰冷的4％多聚甲醛（PFA）的PBS transcardially灌注。 </li><li>灌注后，取出脊髓（SC）和后修复它在4％PFA的PBS在4°CO / N。传送固定​​的SC到PBS中，用30％蔗糖，并保持它为2 - 3天，直到它下沉到管的底部。 </li><li>使用低温恒温器切断脊髓成30微米厚的切片并顺序将它们放置在明胶包被的载玻片或在一个48孔板用0.01M PBS中。 </li></ol></li><li>尼氏染色:要检查所造成的PT伤害进行尼氏staini纳克上脊髓切片如前所述17-20。 <ol><li>简单地说，收集玻片五分之一的脊髓切片和染色用0.25％甲酚紫。取染色的切片（ 图4）的图像。 </li></ol></li><li>免疫组化:正如先前使用浮动部分的方法17,18,20描述。 <ol><li>兔抗的NeuN抗体（1:300），和兔抗:简单地说，通过将O / N在4℃下用兔抗神经胶质纤维酸性蛋白（GFAP）多克隆抗体（300 1）染色脊髓切片Iba1抗体（1:500），随后加入驴抗兔的Alexa 568缀合的抗体（1:400），用于在室温下4小时的二次抗体。采取用荧光显微镜（ 图5）的图像。 </li></ol></li></ol>

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The authors have nothing to disclose.

在这项研究中，我们描述了SC缺血的光化学模型。由于进展遗传工程一直存在于市售的转基因小鼠的激增已经使得有可能研究涉及在SC缺血病理生理学特异性基因的影响。这项研究的目的是开发脊髓缺血可重现小鼠模型。在这里，我们适应皮质PT型号诱导脊髓损伤的小鼠。手术后脊柱静脉和毛细血管对小鼠的背侧处的T11胸椎水平被暴露。然后RB，市售的光敏染料，通过眶后窦路线被注入...

外伤性脊髓损伤（SCI）是影响SC的感觉和植物神经功能毁灭性的临床情况。患者存活SCI往往留下衰弱截瘫的显著影响他们的日常活动和生活1的质量。实验SCI车型已经在科学考察中不可或缺的工具来了解脊髓损伤的病理生理学和相关的神经修复过程。这些模型也被用于测试的目的是在功能恢复各种实验神经保护干预的临床前功效。目前，大部分车型SCI在实践应用中使用的物理钝力机械破坏和伤害的SC。这些方法包括SC 2挫伤，压缩，错位和横断。有人建议，该一次侧机械损伤的缺血集形式的继发性损伤在受伤的SC后<html> 3,4。继发缺血的病因包括广泛组织变性，脑实质出血，有时被阻塞的血管通过组织水肿5-7。作为继发性损伤的SC的完整性被进一步影响的结果，神经元和胶质细胞严重受损功能和活力和凋亡导致在损伤的慢性阶段对梗死增长，类似于缺血半影中风后的生长8,9。几种机制一样兴奋性中毒，自由基的产生，和炎症已经报道负责缺血性细胞死亡以下脊髓损伤10,11。此外，SC缺血是胸腹主动脉瘤修复手术，往往导致截瘫的病人12,13的严重并发症。尽管有如此高的临床影响，目前可脊髓缺血具有高重复性非常少的机型。 核苷酸"&gt; Photothrombosis（PT）是一种常用的方法，在大脑中14-20局灶性缺血的诱导。该技术是相当的非侵入性的，高度可再现的，并产生一个精确的局灶性缺血病灶在脑17的露出面积-21。这是通过光敏染料像玫瑰红（RB）16-20,22或赤藓红的B全身给药达到23接着血管适当光源局部照射。染料的光敏化引起的自由基的生成而扰乱平滑血管内皮的完整性，并引起血小板聚集，随后形成血栓。血流的阻塞由血栓导致在由容器24供给的区域的梗塞。由于减轻控制对强度和辐照的持续时间这一步骤产生高度均匀和可再现的梗塞。此外，这种方法可用于诱导梗死T对于不同的解剖位置，使缺血的作用空间（ 例如 ，灰质与白质）的理解。 目前研究的目的是开发SC缺血小鼠容易和高度可再现的模式。我们描述SC缺血的小鼠模型的PT的过程。结果，从组织学和免疫证明，PT能有效诱导SC梗死，神经元缺失和胶质增生反应。

<table border="0" cellpadding="0" cellspacing="0" width="1050">
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		<col />
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	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">&#160;Rose Bengal</td>
			<td style="width:233px;">Sigma-Aldrich</td>
			<td style="width:180px;">330000</td>
			<td style="width:421px;">20 mg/ml in sterile saline</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">C57Bl6/J</td>
			<td>Jackson lab</td>
			<td>664</td>
			<td>22-25g</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ketamine&#160;</td>
			<td>VEDCO</td>
			<td>NDC-50989-996-06</td>
			<td>100 mg/ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Xylazine&#160;</td>
			<td>VEDCO</td>
			<td>NDC-50989-234-11</td>
			<td>100 mg/ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Betadine solution</td>
			<td>Purdue</td>
			<td>NDC-67618-150-01</td>
			<td>10% povidone iodine topical solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Normal saline</td>
			<td>Abott Laboratories</td>
			<td>04930-04-10</td>
			<td>For diluting RB, anaesthesia and for preventing tissue from drying</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Artificial tears ointment&#160;</td>
			<td>Rugby</td>
			<td>NDC-0536-6550-91</td>
			<td>83% white petrolatum</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ethanol</td>
			<td>Decon labs.Inc</td>
			<td>2716</td>
			<td>70% ethanol for disinfection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Metal halide lamp</td>
			<td>EXFO, Canada</td>
			<td>X-Cite 120 PC&#160;</td>
			<td>Set power at 12%</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Spring scissors&#160;</td>
			<td>Fine Science Tool</td>
			<td>15000-10</td>
			<td>for minor dissection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scissors (angled to side)</td>
			<td>Fine Science Tool</td>
			<td>14063-011</td>
			<td>No. 3 handle</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Standard scalpel</td>
			<td>Fine Science Tool</td>
			<td>10003-12</td>
			<td>for removing muscle</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scalpel blade</td>
			<td>Feather</td>
			<td>2976</td>
			<td>No. 10</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Forceps (curved)</td>
			<td>Fine Science Tool</td>
			<td>11150-10</td>
			<td>for holding tissue</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Forceps (straight)</td>
			<td>Fine Science Tool</td>
			<td>11151-10</td>
			<td>for holding tissue</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Needle holder&#160;</td>
			<td>Fine Science Tool</td>
			<td>12002-12</td>
			<td>for suturing</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tissue adhesive glue</td>
			<td>3M Vetbond</td>
			<td>1469SB</td>
			<td>to adhere to edges of the cut skin</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Monofilament polypropylene&#160;</td>
			<td>USSC Sutures</td>
			<td>VP-521</td>
			<td>Size = 4-0 (for fascia)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Perma-hand silk</td>
			<td>Ethicon</td>
			<td>683G</td>
			<td>Size = 4-0 (for skin)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro drill</td>
			<td>Roboz Surgical Instrument Co. Inc.</td>
			<td>RS-6300</td>
			<td>with bone polishing drill bit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Laser doppler flowmeter</td>
			<td>Moor Instruments</td>
			<td>moorVMS-LDF1</td>
			<td>for monitoring change in blood flow</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Heating pad</td>
			<td>Fine Science Tool</td>
			<td>21052-00</td>
			<td>to prevent hypothermia</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lab-Jack</td>
			<td>Fisher scientific&#160;</td>
			<td>14-673-50</td>
			<td>4x4 in plate to adjust the height of the animal</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">X-Y gliding stage&#160;</td>
			<td>Amscope</td>
			<td>GT100</td>
			<td>for positioning the animal under microscope&#160;&#160;</td>
		</tr>
	</tbody>
</table>

photothrombosis-induced focal ischemia as a model of spinal cord injury in mice

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) below the site of injury. The secondary ischemia that develops following the initial mechanical insult is a serious complication of the SCI and severely impairs the function and viability of surviving neuronal and non-neuronal cells in the SC. In addition, ischemia is also responsible for the growth of lesion during chronic phase of injury and interferes with the cellular repair and healing processes. Thus there is a need to develop a spinal cord ischemia model for studying the mechanisms of ischemia-induced pathology. Focal ischemia induced by photothrombosis (PT) is a minimally invasive and very well established procedure used to investigate the pathology of ischemia-induced cell death in the brain. Here, we describe the use of PT to induce an ischemic lesion in the spinal cord of mice. Following retro-orbital sinus injection of Rose Bengal, the posterior spinal vein and other capillaries on the dorsal surface of SC were irradiated with a green light resulting in the formation of a thrombus and thus ischemia in the affected region. Results from histology and immunochemistry studies show that PT-induced ischemia caused spinal cord infarction, loss of neurons and reactive gliosis. Using this technique a highly reproducible and relatively easy model of SCI in mice can be achieved that would serve the purpose of scientific investigations into the mechanisms of ischemia induced cell death as well as the efficacy of neuroprotective drugs. This model will also allow exploration of the pathological changes that occur following SCI in live mice like axonal degeneration and regeneration, neuronal and astrocytic Ca2+ signaling using two-photon microscopy.

本研究的目的是为了产生在使用PT模型小鼠脊髓缺血。骨脊髓（T10 - T12）以上的所希望的区域之后被减薄，玫瑰红通过眶后窦途径注射，和局部缺血诱导的PT 图1A，B示出的鼠标放置在一个特制的手术。在手术过程中的平台。将小鼠保持在适当位置由口鼻部夹钳和两个可调脊椎动物夹具来稳定脊髓图1C示出了减薄窗口T10的脊髓以上- T12。主要血管及其分支可以清楚地显现。以...

Watch this Scientific Journal Video about Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice at JoVE.com

Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice

Photothrombosis is a minimally invasive and highly reproducible procedure to induce focal ischemia in the spinal cord and serves as a model of spinal cord injury in mice.

Photothrombosis引起的局部脑缺血脊髓损伤的小鼠模型

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) ...

photothrombosis-induced-focal-ischemia-as-model-spinal-cord-injury

Research

JoVE Journal

Medicine

13.2K 次观看.  University of Missouri. Photothrombosis is a minimally invasive and highly reproducible procedure to induce focal ischemia in the spinal cord and serves as a model of spinal cord injury in mice. 

视频: Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

在大鼠脊髓损伤后的急性和慢性触觉测试

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

科研

医学

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

控制宫颈裂伤损伤小鼠

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

A Contusion Model of Severe Spinal Cord Injury in Rats

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is described to obtain a consistent model of severe SCI. Use of a stereotactic frame and computer controlled impactor allows for creation of reproducible injury. Hypothermia and urinary tract infection pose significant challenges in the post-operative period. Careful monitoring of animals with daily weight recording and bladder expression allows for early detection of post-operative complications. The functional results of this contusion model are equivalent to transection models. The contusion model can be utilized to evaluate the efficacy of both neuroprotective and neuroregenerative approaches.

A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

挫伤模型脊髓严重损伤的保护作用

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is ...

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often unreported mortality rates that may confound analyses. Bilateral renal pedicle clamping is commonly used to induce IR-AKI, but differences between effective clamp pressures and/or renal responses to ischemia between kidneys often lead to more variable results. In addition, shorter clamp times are known to induce more variable tubular injury, and while mice undergoing bilateral injury with longer clamp times develop more consistent tubular injury, they often die within the first 3 days after injury due to severe renal insufficiency. To improve post-injury survival and obtain more consistent and predictable results, we have developed two models of unilateral ischemia-reperfusion injury followed by contralateral nephrectomy. Both surgeries are performed using a dorsal approach, reducing surgical stress resulting from ventral laparotomy, commonly used for mouse IR-AKI surgeries. For induction of moderate injury BALB/c mice undergo unilateral clamping of the renal pedicle for 26 min and also undergo simultaneous contralateral nephrectomy. Using this approach, 50-60% of mice develop moderate AKI 24 hr after injury but 90-100% of mice survive. To induce more severe AKI, BALB/c mice undergo renal pedicle clamping for 30 min followed by contralateral nephrectomy 8 days after injury. This allows functional assessment of renal recovery after injury with 90-100% survival. Early post-injury tubular damage as well as post injury fibrosis are highly consistent using this model.

We describe models of moderate and severe ischemia-reperfusion-induced kidney injury in which the mice undergo unilateral renal pedicle clamping followed by simultaneous or delayed contralateral nephrectomy, respectively. These models consistently give rise to renal dysfunction and post-injury fibrosis, but injury severity and survival are dependent on mouse background, age and surgical equipment.

急性肾损伤和伤后纤维化小鼠缺血再灌注模型

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

Reproducable Paraplegia by Thoracic Aortic Occlusion in a Murine Model of Spinal Cord Ischemia-reperfusion

Background 
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered advancements to decrease the occurrence this injury.&#160;The current model demonstrates reproducible lower&#160;extremity paralysis following thoracic aortic occlusion.
Methods 
Adult male C57BL6 mice were anesthetized with isoflurane. Through a cervicosternal incision the aorta was exposed. The descending thoracic aorta and left subclavian arteries were identified without entrance into pleural space. Skeletonization of these arteries was followed by immediate closure (Sham) or occlusion for 4 min (moderate ischemia) or 8 min (prolonged ischemia). The sternotomy and skin were closed and the mouse was transferred to warming bed for recovery.&#160; Following recovery, functional analysis was obtained at 12 hr intervals until 48 hr.
Results 
Mice that underwent sham surgery showed no observable hind&#160;limb deficit. Mice subjected to moderate ischemia for 4 min had minimal functional deficit at 12 hr followed by progression to complete paralysis at 48 hr. Mice subjected to prolonged ischemia had an immediate paralysis with no observable hind-limb movement at any point in the postoperative period. There was no observed intraoperative or post&#160;operative mortality.
Conclusion 
Reproducible lower extremity paralysis whether immediate or delayed can be achieved in a murine model. Additionally, by using a median sternotomy and careful dissection, high survival rates, and reproducibility can be achieved.

The lack of mechanistic understanding of spinal cord ischemia-reperfusion injury has hindered further adjuncts to prevent paraplegia following high risk aortic operations. Thus, the development of animal models is imperative. This manuscript demonstrates reproducible lower extremity paralysis following thoracic aortic occlusion in a murine model.

可重放截瘫胸主动脉阻断脊髓缺血再灌注的小鼠模型

Background
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered ...

Controlled Cortical Impact Model for Traumatic Brain Injury

Every year over a million Americans suffer a traumatic brain injury (TBI). Combined with the incidence of TBIs worldwide, the physical, emotional, social, and economical effects are staggering. Therefore, further research into the effects of TBI and effective treatments is necessary. The controlled cortical impact (CCI) model induces traumatic brain injuries ranging from mild to severe. This method uses a rigid impactor to deliver mechanical energy to an intact dura exposed following a craniectomy. Impact is made under precise parameters at a set velocity to achieve a pre-determined deformation depth. Although other TBI models, such as weight drop and fluid percussion, exist, CCI is more accurate, easier to control, and most importantly, produces traumatic brain injuries similar to those seen in humans. However, no TBI model is currently able to reproduce pathological changes identical to those seen in human patients. The CCI model allows investigation into the short-term and long-term effects of TBI, such as neuronal death, memory deficits, and cerebral edema, as well as potential therapeutic treatments for TBI.

Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be performed.

控制皮层影响模型创伤性脑损伤

Every year over a million Americans suffer a traumatic brain injury (TBI).  Combined with the incidence of TBIs worldwide, the physical, emotional, ...

A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter connected to a radio transmitter into the femoral artery of rats that underwent a T4 spinal cord transection with or without grafting of embryonic brainstem-derived neural stem cells expressing green fluorescent protein. Compared to other methods such as cannula insertion or tail-cuff, telemetry is advantageous to continuously monitor blood pressure and heart rate in freely moving animals. It is also capable of long term multiple data acquisitions. In spinal cord injured rats, basal cardiovascular data under unrestrained condition and autonomic dysreflexia in response to colorectal distension were successfully recorded. In addition, cardiovascular parameters before and after SCI can be compared in the same rat if a transmitter is implanted before a spinal cord transection. One limitation of the described telemetry procedure is that implantation in the femoral artery may influence the blood supply to the ipsilateral hindlimb.

We present a protocol for using a radio-telemetric system to record cardiovascular parameters in T4 spinal cord transected rats eight weeks after embryonic brainstem neural stem cell grafting into the lesion site. Telemetry is an advanced technique to accurately evaluate cardiovascular function in conscious freely moving spinal cord injured rats.

无线电遥测系统，以监测大鼠脊髓横断和胚胎神经干细胞移植在心血管功能

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter ...

Neural Stem Cell Transplantation in Experimental Contusive Model of Spinal Cord Injury

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can be used both to investigate the biological responses to injury and to test potential therapies. Contusion or compression injury delivered to the surgically exposed spinal cord are the most widely used models of the pathology. In this report the experimental contusion is performed by using the Infinite Horizon (IH) Impactor device, which allows the creation of a reproducible injury animal model through definition of specific injury parameters. Stem cell transplantation is commonly considered a potentially useful strategy for curing this debilitating condition. Numerous studies have evaluated the effects of transplanting a variety of stem cells. Here we demonstrate an adapted method for spinal cord injury followed by tail vein injection of cells in CD1 mice. In short, we provide procedures for: i) cell labeling with a vital tracer, ii) pre-operative care of mice, iii) execution of a contusive spinal cord injury, and iv) intravenous administration of post mortem neural precursors. This contusion model can be utilized to evaluate the efficacy and safety of stem cell transplantation in a regenerative medicine approach.

Spinal cord injury is a traumatic condition that causes severe morbidity and high mortality. In this work we describe in detail a contusion model of spinal cord injury in mice followed by a transplantation of neural stem cells.

在脊髓损伤的实验挫伤型号神经干细胞移植

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can ...

Calibrated Forceps Model of Spinal Cord Compression Injury

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The calibrated forceps model of compression injury is a convenient, low cost, and very reproducible animal model for SCI. We used a pair of modified forceps in accordance with the method published by Plemel et al. (2008) to laterally compress the spinal cord to a distance of 0.35 mm. In this video, we will demonstrate a dorsal laminectomy to expose the spinal cord, followed by compression of the spinal cord with the modified forceps. In the video, we will also address issues related to the care of paraplegic laboratory animals. This injury model produces mice that exhibit impairment in sensation, as well as impaired hindlimb locomotor function. Furthermore, this method of injury produces consistent aberrations in the pathology of the SCI, as determined by immunohistochemical methods. After watching this video, viewers should be able to determine the necessary supplies and methods for producing SCI of various severities in the mouse for studies on SCI and/or treatments designed to mitigate impairment after injury.

Spinal cord injury models should be highly reproducible. We demonstrate that the calibrated forceps compression model of spinal cord injury is an easy to use surgical method for generating reproducible injuries to the murine spinal cord.

脊髓压迫损伤的校准钳型

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The ...

A Neonatal Mouse Spinal Cord Compression Injury Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal cord. A major current area of research is focused on the mechanisms of adaptive plasticity that underlie spontaneous or induced functional recovery following SCI. Spontaneous functional recovery is reported to be greater early in life, raising interesting questions about how adaptive plasticity changes as the spinal cord develops. To facilitate investigation of this dynamic, we have developed a SCI model in the neonatal mouse. The model has relevance for pediatric SCI, which is too little studied. Because neural plasticity in the adult involves some of the same mechanisms as neural plasticity in early life1, this model may potentially have some relevance also for adult SCI. Here we describe the entire procedure for generating a reproducible spinal cord compression (SCC) injury in the neonatal mouse as early as postnatal (P) day 1. SCC is achieved by performing a laminectomy at a given spinal level (here described at thoracic levels 9-11) and then using a modified Yasargil aneurysm mini-clip to rapidly compress and decompress the spinal cord. As previously described, the injured neonatal mice can be tested for behavioral deficits or sacrificed for ex vivo physiological analysis of synaptic connectivity using electrophysiological and high-throughput optical recording techniques1. Earlier and ongoing studies using behavioral and physiological assessment have demonstrated a dramatic, acute impairment of hindlimb motility followed by a complete functional recovery within 2 weeks, and the first evidence of changes in functional circuitry at the level of identified descending synaptic connections1.

This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

新生儿小鼠脊髓压迫损伤模型

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...