我们感谢埃默里大学部兽医照顾动物资源的工作人员。资金来源包括：ALS协会，国防部，Neuralstem公司

1。动物用已批准程序演示本文由埃默里大学实验动物护理和使用委员会（IACUC）。女哥廷根小型猪，体重约15-20公斤。 2。麻醉动物约12小时，在手术前禁食。动物镇静和麻醉诱导肌注氯氨酮（35毫克/千克），乙酰丙嗪（1.1毫克/公斤），与阿托品（0.04毫克/千克）的鸡尾酒。动物的氧和1-3％的异氟醚全身麻醉气管插管和维护。在这一点上，每个动物的背部和头部被剃光。麻醉深度监测的兽医人员。缺乏叉，角膜和眼睑反射消失，以及心脏和呼吸频率，脉搏血氧饱和度，直接/间接血压，呼气末二氧化碳测量，肌张力/对有害刺激的反应是用来监测深度<html>麻醉。 3。定位动物被带到手术室，置于俯卧在一个框架定制设计的模仿患者定位在杰克逊脊柱外科表。帧利用可调吊索，划归的胸部和骨盆的动物，使腹部挂自由，因此，尽量减少腹部和胸部，和随之而来的的硬膜外静脉出血（ 图1）上的压力。该框架还提供了外固定脊柱的步骤1。 此外，动物被放置在温水重新循环垫，以保持身体的温度和耳缘静脉插管输液和任何手术过程中提供必要的药物。最后，外科领域prepped用酒精和Chlorhexadine或聚维酮碘溶液，手术布被放置在外科领域。 文件“&gt; 4。椎板切除术约10-15厘米的皮肤切口和剥离椎旁肌肉组织的脊柱双边。接着，背侧多级椎板切除术进行。三椎覆C3-C5，L2-L4段的椎板和棘突咬骨钳和手术钻去除。 5。放置在脊髓德里克 - 我们称之为脊柱井架设备的设计，交付（基因和细胞）治疗脊髓2-5。此设备的设计和改进的详细讨论可以找到Riley 等人 ，2011。 为了确保移动设备以病人，经皮员额放置通过1厘米皮肤切口的上方和下方的主切口和安装在上面和下面的主切口椎板。 接着，两个集成的牵开器被连接到了聚焦回合经皮职位上方和下方的切口部位的脊柱的区域已经经历椎板切除术暴露。 6。硬脑膜开幕与援助的的伍德森牙科工具和一个11刀片服务器，一个2.5厘米的切口通过硬脑膜，暴露脊髓。硬膜反射离开的脑膜层使用4-0 Nurolon的缝合，固定深椎旁肌肉组织。 外科手术的肉饼被放置在喙和尾部的极端开幕。这些提供了部分障碍，脑脊液流动的外科医生，不要损坏电线将吸盘还提供了一个安全的目标。在人类中，在手术显微镜下放大，软膜表面解剖在这一点上。由于技术上的限制，这个过程并不需要或可行的动物。 7。脊髓注射和侧向位移的脊髓德里克额外注资小号紧接注射，静脉推注甲基强的松龙（125毫克，IV），以防止脊髓肿胀。 在这一点上，该平台轨道系统的连接和调整，以适应相应的长度的侧轨。 Gondola的 顶部加载到2个酒吧和Z驱动器被安装在万向接头。接着，被加载的插管放置到微硬盘。显微注射平台上使用的万向接头，冠状面和矢状面的角度进行调整，以确保脊髓的注射剂按照安置在插管的表面正交的轨迹。针定位相应的内侧背根进入区（DREZ）。 DREZ被确定在3.5倍手术放大镜的放大倍率，渗透在一个点线面垂直轨迹&lt;1 mm的内侧。 在人类中，手术前的MRI提供了一个基线评估Øf的脊髓尺寸为手术计划。此外，测量厚度的脊髓腹侧角来确定的目标深度。 将悬浮液注入在从软膜接触的深度为4毫米。的凸缘由ULTEM塑料制成的作为软表面上的挡块，以防止针推进深度超过所需。一旦被定位在目标的针尖，刚性的金属外套筒被拉升，留下暴露的柔性管。注射完毕后，将针留在原地，一个额外的1分钟，以防止细胞回流上升插管注入道。 并小心避免表面血管的微硬盘无论是偏侧或rostro的，尾部略作调整。从普及率的网站中有一些出血现象可能会出现。当遇到这种出血，微肉饼被放置在穿刺部位出血和吸力被施加到它们以毛细血满分插管penetrATION的网站，防止积聚在电源线。这可靠地允许血液凝结。烧灼避免直接的压力。 针去除后，立体定位仪沿rostro尾椎轴迁移到下一个目标网站，由2个或4毫米或必要的，以避免可见的血管，脊髓的背侧面分开。这个过程被重复多次，在一个给定的研究提出。 8。浮动套管使用一个自定义的输液套管的直径缩小为注射剂。所述套管包括30隔距柔性可变长度的硅橡胶管连接到固定长度的30隔距的斜口针。的前端部嵌合，其被安装到一个微量注射器泵与一个Hamilton鲁尔锁定。近端的硅橡胶管ensheathed内24隔距刚性的外套管，座椅上的注射针的近端凸缘。该法兰盘座位外套管和作为用于注射针的深度停止。为每次注射，输注的治疗的悬浮液的适当体积的由使用一个预先校准MINJ-PD的微量注射器泵（的Tritech研究公司，洛杉矶，CA）的在5微升每分钟的速率。 9。关闭一旦所有的注射剂，轻轻地去除脊髓井架和关闭切口在四层。关闭硬脑膜使用一个4.0 Nurolon的针，水密性方式。 0薇乔缝线用于深达肌层。 0薇乔缝线也水密性方式，筋膜，然后关闭。真皮层最终收报2.0薇乔，与正在运行的线迹。皮肤缝合完成后，用3-0尼龙线。 10。恢复和疼痛管理动物被拔除气管插管和监测麻醉后2小时恢复。接着，动物被转移到各个C年龄和用于食品消费，排便，排尿，每天至少一次监测。 疼痛管理，经皮芬太尼贴剂（75微克）装订在动物背部手术后镇痛三天。此外，也可以给予丁丙诺啡（0.05毫克/千克，BID，IM），最多至三天后可操作。 11。结果有代表性的结果临床和行为观察手术前，然后记录在1〜7天，每周根据研究设计，直到终点。行为数据被收集评估神经系统的发病率，如先前描述的6。感觉功能的评估是通过存在或不存在的戒断反应机械刺激的前部和后肢脚趾。运动功能如下Tarlov评分（ 表1）：0 -麻痹，没有动静; 1 -能感觉到强直性后肢轻微米; 2 - ovement后肢的运动，但无法坐或站，3 - 站立和行走，但共济失调和短期内的能力; 4 - 完全恢复，运动功能正常。 被确定的程序的安全能力的动物返回到手术前的基线。瞬态的神经功能缺损应主要解决手术后1和7天之间，取决于动物的品种和程序（注射数的一些变体中，除其他的参数）。永久的发病率被定义为持久的神经功能障碍，不解决的时候，动物达到的IACUC默认端点（ 图2）。

<ol>
	<li>Usvald, D., 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=Analysis+of+dosing+regimen+and+reproducibility+of+intraspinal+grafting+of+human+spinal+stem+cells+in+immunosuppressed+minipigs.">Analysis of dosing regimen and reproducibility of intraspinal grafting of human spinal stem cells in immunosuppressed minipigs.</a> Cell Transplant. 19, 1103-1122 (2010).</li><li>Riley, J., 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=Targeted+spinal+cord+therapeutics+delivery:+stabilized+platform+and+microelectrode+recording+guidance+validation.">Targeted spinal cord therapeutics delivery: stabilized platform and microelectrode recording guidance validation.</a> Stereotact. Funct. Neurosurg. 86, 67-74 (2008).</li><li>Federici, T., Riley, J., Park, J., Bain, M., Boulis, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preclinical+safety+validation+of+a+stabilized+viral+vector+direct+injection+approach+to+the+cervical+spinal+cord.">Preclinical safety validation of a stabilized viral vector direct injection approach to the cervical spinal cord.</a> Clin. Transl. Sci. 2, 165-167 (2009).</li><li>Riley, J. P., Raore, B., Taub, J. S., Federici, T., Boulis, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Platform+and+Cannula+Design+Improvements+for+Spinal+Cord+Therapeutics+Delivery.">Platform and Cannula Design Improvements for Spinal Cord Therapeutics Delivery.</a> Neurosurgery. 69, 147-154 (2011).</li><li>Riley, J., 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=Cervical+spinal+cord+therapeutics+delivery:+preclinical+safety+validation+of+a+stabilized+microinjection+platform.">Cervical spinal cord therapeutics delivery: preclinical safety validation of a stabilized microinjection platform.</a> Neurosurgery. 65, 754-761 (2009).</li><li>Raore, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cervical+multilevel+intraspinal+stem+cell+therapy:+assessment+of+surgical+risks+in+Gottingen+minipigs.">Cervical multilevel intraspinal stem cell therapy: assessment of surgical risks in Gottingen minipigs.</a> Spine. 36, 164-171 (2011).</li><li>Boulis, N. 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=Translational+stem+cell+therapy+for+amyotrophic+lateral+sclerosis.">Translational stem cell therapy for amyotrophic lateral sclerosis.</a> Nat. Rev. Neurol. , (2011).</li><li>Riley, J., 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=Intraspinal+Stem+Cell+Transplantation+in+ALS:+A+Phase+I+Safety+Trial,+Technical+Note+&#38;+Interim+Safety+Outcomes.">Intraspinal Stem Cell Transplantation in ALS: A Phase I Safety Trial, Technical Note &#38; Interim Safety Outcomes.</a> Neurosurgery. 71 (2), 405-416 (2012).</li><li>Boulis, N. 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=Lumbar+intraspinal+injection+of+neural+stem+cells+in+patients+with+ALS:+results+of+a+Phase+I+trial+in+12+patients.">Lumbar intraspinal injection of neural stem cells in patients with ALS: results of a Phase I trial in 12 patients.</a> Stem Cells. 30 (6), 1144-1151 (2012).</li></ol>

Boulis是博士发明的设备，使安全，准确的人类脊髓注射。 Neuralstem，公司已经购买了这项技术的独家授权。 Boulis的清查博士占这项费用和有权利分配这项技术的专利使用费。其他作者有没有透露。

尽管人类的7-9所描述的技术进行批准，为了脊髓治疗成功的关键问题仍有待回答。 脊髓耐受性脑实质内注射需要一个严格的理解，使开发疗法的试验性脱髓鞘性，变性，外伤性脊髓病的规划和执行。目前，还没有清晰的认识注射的数量，大型哺乳动物的脊髓可以承受的，没有暂时性和永久性的发病率。同样，注射的间隔可能是影响发病率。此外，宏观的（ 例如 ，通风相关?...

<table border="1"><tbody><tr><td> 的设备名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td></tr><tr><td>脊柱德里克 - </td><td> Neuralstem公司</td><td></td><td> Neuralstem，公司已经购买了这项技术的独家授权</td></tr><tr><td> MINJ-PD微量注射器泵</td><td> ，Tritech公司研究公司</td><td></td><td>这个特定的应用程序定制</td></tr><tr><td>浮动套管</td><td> Neuralstem公司</td><td></td><td>自定义设计的这个特定的应用程序的</td></tr></tbody></table>

surgical technique for spinal cord delivery of therapies: demonstration of procedure in gottingen minipigs

这是一个紧凑的可视描述的手术技术和装置，用于提供到脊髓（基因和细胞）疗法的组合。虽然该技术被证明在动物体内，该过程是FDA批准的，目前用于ALS患者的脊髓干细胞移植到。虽然FDA已确认疗效高度特征的啮齿动物模型中证明的原则上的数据，大型动物的使用被认为是关键的，用于验证的组合的外科手术过程中，移动设备，和供人类使用的安全性的最终治疗。的大小，解剖学和一般的脆弱性脊椎和脊椎神经的猪被确认为更好的模型的人。而且从人类的揭露和处理以及封闭伤口在猪脊髓，手术过程中几乎没有区别。我们相信，健康的猪模型是一个关键校准第一步在研究程序安全。

<table border="1"><tbody><tr><td colspan="2"> Tarlov评分</td></tr><tr><td> 0 </td><td>瘫痪，无运动</td></tr><tr><td> 1 </td><td>能感觉到的强直性在后肢，轻微的运动</td></tr><tr><td> 2 </td><td>在后肢的运动，但不能坐或站</td></tr><tr><td> 3 </td><td>站立和行走的能力，但共济失调，短期内</td></tr><tr><td> 4 </td><td>完全恢复，运动功能正常; </td></tr></tbody></table> ?...

Watch this Scientific Journal Video about Surgical Technique for Spinal Cord Delivery of Therapies: Demonstration of Procedure in Gottingen Minipigs at JoVE.com

Surgical Technique for Spinal Cord Delivery of Therapies: Demonstration of Procedure in Gottingen Minipigs

短期视觉描述的手术技术和设备用于交付（基因和细胞）治疗脊髓。中的动物，但该技术被证明是完全平移和当前正用于人类应用。

外科技术治疗脊髓交付：在哥廷根小型猪演示程序

This is a  compact visual description of a combination of surgical technique and device  for the  delivery of (gene and cell) therapies into the spinal ...

surgical-technique-for-spinal-cord-delivery-therapies-demonstration

Research

JoVE Journal

Medicine

15.2K 次观看.  Emory University. 短期视觉描述的手术技术和设备用于交付（基因和细胞）治疗脊髓。中的动物，但该技术被证明是完全平移和当前正用于人类应用。

视频: Surgical Technique for Spinal Cord Delivery of Therapies: Demonstration of Procedure in Gottingen Minipigs

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

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 ...

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 ...

Minimal Invasive Surgical Procedure of Inducing Myocardial Infarction in Mice

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last decades. For clarification of the underlying mechanisms and the development of new therapeutic strategies, the availability of valid animal models are mandatory. Since we need new insights into pathomechanisms of cardiovascular diseases under in vivo conditions to combat myocardial infarction, the validity of the animal model is a crucial aspect. However, protection of animals are highly relevant in this context. Therefore, we establish a minimally invasive and simple model of myocardial infarction in mice, which assures a high reproducibility and survival rate of animals. Thus, this models fulfils the requirements of the 3R principle (Replacement, Refinement and Reduction) for animal experiments and assure the scientific information needed for further developing of therapeutical strategies for cardiovascular diseases.

A highly reproducible model for myocardial infarction in mice with minimal invasive manipulations is described. The model can be easily performed, resulting in a high reproducibility and survival rate. Thus, the described model will reduce the number of required animals as requested by the 3R principle (Replacement, Refinement and Reduction).

在诱导小鼠心肌梗死的微创手术过程

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last ...

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 ...

Contrast Enhanced Ultrasound Imaging for Assessment of Spinal Cord Blood Flow in Experimental Spinal Cord Injury

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an important target for neuroprotective therapies. Although several techniques have been described to assess SCBF, they all have significant limitations. To overcome the latter, we propose the use of real-time contrast enhanced ultrasound imaging (CEU). Here we describe the application of this technique in a rat contusion model of SCI. A jugular catheter is first implanted for the repeated injection of contrast agent, a sodium chloride solution of sulphur hexafluoride encapsulated microbubbles. The spine is then stabilized with a custom-made 3D-frame and the spinal cord dura mater is exposed by a laminectomy at ThIX-ThXII. The ultrasound probe is then positioned at the posterior aspect of the dura mater (coated with ultrasound gel). To assess baseline SCBF, a single intravenous injection (400 &#181;l) of contrast agent is applied to record its passage through the intact spinal cord microvasculature. A weight-drop device is subsequently used to generate a reproducible experimental contusion model of SCI. Contrast agent is re-injected 15 min following the injury to assess post-SCI SCBF changes. CEU allows for real time and in-vivo assessment of SCBF changes following SCI. In the uninjured animal, ultrasound imaging showed uneven blood flow along the intact spinal cord. Furthermore, 15 min post-SCI, there was critical ischemia at the level of the epicenter while SCBF remained preserved in the more remote intact areas. In the regions adjacent to the epicenter (both rostral and caudal), SCBF was significantly reduced. This corresponds to the previously described &#8220;ischemic penumbra zone&#8221;. This tool is of major interest for assessing the effects of therapies aimed at limiting ischemia and the resulting tissue necrosis subsequent to SCI.

Contrast Enhanced Ultrasound imaging is a reliable in-vivo tool for quantifying spinal cord blood flow in an experimental rat spinal cord injury model. This paper contains a comprehensive protocol for application of this technique in association with a contusion model of thoracic spinal cord injury.

超声造影成像脊髓血流量的评估实验脊髓损伤

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an ...

The Mouse Round-window Approach for Ototoxic Agent Delivery: A Rapid and Reliable Technique for Inducing Cochlear Cell Degeneration

Investigators have utilized a wide array of animal models and investigative techniques to study the mammalian auditory system. Much of the basic research involving the cochlea and its associated neural pathways entails exposure of model cochleae to a variety of ototoxic agents. This allows investigators to study the effects of targeted damage to cochlear structures, and in some cases, the self-repair or regeneration of those structures. Various techniques exist for delivery of ototoxic agents to the cochlea. When selecting a particular technique, investigators must consider a number of factors, including the induction of inadvertent systemic toxicity, the amount of cochlear damage produced by the surgical procedure itself, the type of lesion desired, animal survivability, and reproducibility/reliability of results. Currently established techniques include parenteral injection, intra-peritoneal injection, trans-tympanic injection, endolymphatic sac injection, and cochleostomy with perilymphatic perfusion. Each of these methods has been successfully utilized and is well described in the literature; yet, each has various shortcomings. Here, we present a technique for topical application of ototoxic agents directly to the round window niche. This technique is non-invasive to inner ear structures, produces rapid onset of reliably targeted lesions, avoids systemic toxicity, and allows for an intra-animal control (the contra-lateral ear). Results stemming from this approach have helped deeper understanding of auditory pathophysiology, cochlear cell degeneration, and regenerative capacity in response to an acute injury. Future investigations may use this method to conduct interventional studies involving gene therapy and stem cell transplantation to combat hearing loss.

Various methods exist for introducing ototoxic agents to the cochleae of animal models. Presented is a surgical protocol for delivery of ototoxic agents to the round window niche. The procedure is reliable, creates targeted intra-cochlear lesions, and avoids mechanical damage to the microarchitecture. Examination of cochlear self-repair/regeneration is possible.

老鼠圆窗途径耳毒性剂递送：一个快速和可靠的技术诱导耳蜗细胞变性

Investigators have utilized a wide array of animal models and investigative techniques to study the mammalian auditory system. Much of the basic research ...

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 ...

Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via recanalization of the coronary artery is the most effective strategy for reducing the size of ischemic myocardium. The coronary microvasculature cannot be visualized and imaged&#160;in vivo, but there are several invasive and noninvasive techniques that can be used to assess parameters which depend directly on coronary microvascular function. The endothelial function after ischemia reperfusion can be assessed also at the level of the coronary circulation via the coronary flow reserve (CFR).&#160;In this study, peak velocity of left anterior descending (LAD) coronary arteries was measured in rats in vivo via Transthoracic Doppler Echocardiography during resting and stress challenge (induced by Dobutamine). A normal heart can increase its coronary blood flow up to four times above the resting values during stress induction. Following ischemia reperfusion, we found a significantly diminished CFR, which can be used as a marker of coronary microvascular dysfunction. CFR has opened a window on the importance of microvascular dysfunction and has been shown to predict cardiovascular risk independent of whether the severe obstructive disease is present.

Coronary flow reserve (CFR), is defined as the ratio of maximal coronary blood flow to the resting coronary blood flow. We present a protocol for evaluating CFR in rats via ultrasound, which offers the opportunity to predict cardiovascular risk factors in the absence of obstructive coronary disease.

大鼠心肌缺血再灌注后冠状动脉流动储备评价

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via ...