The work was supported by grants from NIH/NINDS (NS054883), Craig H. Neilsen Foundation (280072), and the Veterans Administration and Canadian Spinal Research Organization. We thank the Rat Resource and Research Center, University of Missouri, Columbia, Missouri, for providing GFP rats.

所有动物协议获得批准的机构动物照顾及使用委员会（IACUC）。实验动物照护及安全指引，美国国立卫生研究院严格遵循。动物外科手术得到充分的治疗减少疼痛和不适。 1，脊髓Curgery和细胞移植<ol><li>之前，所有的手术高压灭菌手术器械。用热珠灭菌器（精细的科学工具），以从不同的动物之间的程序删除工具病原体和微生物污染物。在手术过程中使用无菌手术手套，长袍，和窗帘。采用无菌手术技术为每个外科手术。 </li><li>麻醉雌性费希尔344大鼠氯胺酮（25毫克/毫升），甲苯噻嗪（1.3毫克/毫升），以及乙酰丙嗪（0.25毫克/毫升）的一个组合（2毫升/千克）经腹膜内注射（IP）给药。 </li><li>剃须后区和优碘和乙醇反复擦拭皮肤。</li><li>使用＃10刀片切开皮肤并直言不讳地剖析肌肉层。用手术刀分离椎骨，露出T3椎体，并用细尖咬骨钳进行椎板切除术背。 </li><li>切开硬脑膜纵向和使用虹膜切除术剪刀和microaspiration的组合来创建脊髓树桩之间的约1mm rostrocaudal间隙在T4水平横切脊髓。 </li><li>等待大约1 - 2分钟，直到出血停止完全，然后缝合肌肉3-0 Vycril，关闭伤口剪辑皮肤。 </li><li>注入乳酸林格氏液（5毫升）中，叔丁啡（0.035毫克/千克），和氨苄青霉素（33毫克/千克）经皮下立即手术后和维持大鼠在温暖的培养箱中，直到清醒。 </li><li>注入林格氏液和氨苄青霉素溶液每日达10天一次，和丁丙诺啡每日3天或直到疼痛和痛苦的迹象消失两次。不要一个动物返回其他动物，直到f的公司ully恢复。 </li><li>每天大约两个星期手动排空膀胱两次，直到建立反射性膀胱排空，然后在必要时排空膀胱，每天一次整个生存。 </li><li>两周后，如上述那样重新麻醉SCI大鼠并重新露出脊髓病灶部位。保持关闭，以保持植入细胞在病变部位的硬脑膜。 </li><li>注入10微升的细胞溶液（3.5×10 5微升），从E14，无处不在的绿色荧光蛋白转基因大鼠胚胎收集和嵌入在纤维蛋白原和凝血酶5,6，进入病灶腔的震中和病变多注射头端和尾端接口位点，用拉玻璃微量移为40微米的内径，连接到Hamilton注射器。 </li><li>缝合肌肉层，关闭伤口剪辑皮肤。 </li><li> 1.8 - 按照步骤1.6中所述进行手术后注射和膀胱护理。 </li><li>一个星期前灌注，在脊髓7注入0.5％荧光金（FG，0.4毫升蒸馏水）腹膜内给逆行标签交感神经节前神经元。 </li></ol> 2，发射器植入术<ol><li>浸泡transmittersin 2％戊二醛溶液（4毫升50％的戊二醛的96毫升蒸馏水的水）为至少1 - 2小时（最多10小时），在室温下进行灭菌。 </li><li>用无菌的0.9％生理盐水彻底冲洗发射机3次，并将它们存储在盐水中，直到使用（不超过1小时）。 </li><li> E14细胞移植（10周后受伤）八周后，reanesthetize是经历了脊髓损伤有或无细胞移植和天真的对照组老鼠。 </li><li>剃腹部和后肢。请用聚维酮碘皮肤。将大鼠上手术台取仰卧位。 </li><li>切开上使用＃15刀片右侧腹腹部和大腿内侧的皮肤上。 </li><li>通过S切ubcutaneous结缔组织，露出用小剪刀股血管和神经束。 </li><li>分开的静脉和神经用细镊子尖端弯曲股动脉。 </li><li>把三丝缝合动脉的下方，并在各缝合一松结。 </li><li>申请0.1毫升利多卡因（2％）的动脉表面，以引起血管舒张用于随后导尿。 </li><li>近端通过拉伸一个松散的丝线缝合固定远端血管具有永久丝结和临时块。 </li><li>使用20号弯针的穿刺动脉，插入使用导管插入工具的遥测导管（8厘米长）的一角。 </li><li>插入导管吻侧最多到4厘米，从而将前端在胸主动脉。 </li><li>通过捆扎的股动脉周围三丝缝线锚定在血管内的导管。 </li><li>使皮下口袋沿着肋骨和T的尾部边缘之间的侧翼他的膝盖的范围最颅延用钝的剪刀。 </li><li>插入发射机体装入口袋并缝合到周围的发射机，以避免过度的运动的结缔组织。 </li><li>缝合用6号丝线皮肤。 </li><li>注入乳酸林格氏液（5毫升）中，叔丁啡（0.035毫克/千克），和氨苄青霉素（33毫克/千克）经皮下立即手术后和维持大鼠在温暖的培养箱中，直到清醒。 </li></ol> 3，基础平均动脉压（MAP）和心率（HR）拍摄<ol><li>由于早一天以下变送器植入，把一个单一的动物在接收器垫，打开发射器。等待大约10 - 15分钟至，观察者的动物和稳定的心血管参数。 </li><li>记录静息血压和心率，这是来自于脉冲动脉压与计算机数据采集系统至少1小时。收集的数据每5秒。 </li><li>监测的imals的可见痉挛的发生过程中连续，并删除数据点。每只动物的平均数据点，得到平均值。 </li></ol> 4，结直肠扩张引起的自主神经反射异常<ol><li>抑制NSC接枝或脊髓损伤对照组大鼠在里面的食物颗粒发射机接收机提供的毛巾。通常只可以留在合作过程中。 </li><li>插入乳胶气球尖导管插入直肠大约2厘米，它固定到尾部用胶带8。 </li><li>打开发射器，等待10 - 15分钟，使血压恢复到preinsertion基线。 </li><li>诱发结直肠扩张由充气气球慢慢在10秒内用1.4毫升空气1分钟，以产生大约30毫米汞柱的压力。 </li><li>记录血压和心率1分钟前，中1分钟，结直肠扩张后1分钟;样本数据的3分钟过程中，每3秒。 </li><li>执行2 - 每只动物3项试验有两个审判之间至少有15分钟的恢复时间。 </li><li>过量的动物（IP）以上，如果没有进一步的评估描述双倍剂量的麻醉组合。动物灌注生理盐水后4％多聚甲醛。 </li><li>对于每一个动物，之前分别在结直肠扩张平均的值;计算的基准，并为每个审判腹胀引起的血压和心率的变化之间的差异;平均在2 - 3的试验，以获得平均值。 </li></ol> 5，发射器清洗<ol><li>麻醉后但灌注前请清除动物体内发射器。在充满蒸馏水清洗，直到烧杯中浸泡，立即;避免的遥测设备的干燥。 </li><li>的遥测传输到1％的Terg-A-ZYME酶清洗溶液（10g / L的水）中24小时，在室温下进行。 </li><li>葱发射机导管的前端有一个钝的30号针连接到葱注射器。 </li><li>使用折叠软组织仔细擦干发射机，并将其存储在原始塑料托盘。 </li></ol>

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作者什么都没有透露。

传统上，流体填充的套管插入动脉并连接到压力换能器来记录心血管参数在每个动物11的终端快照。连续监测心血管性能很长一段时间，无线电遥测系统被应用在许多实验室。这更精致的工具可以记录在有意识的血压，自由活动动物。相较于充满液体的导管，遥测被认为是一种更先进的技术来准确地评估心血管功能的脊髓损伤动物。虽然在本研究中的发射机被记录之前植入仅1天，它可前?...

脊髓损伤（SCI）在较高水平后心血管功能障碍发生。它表现在无序的血压和心脏率在休息，体位性低血压，运动性低血压及自主神经反射异常响应损伤水平以下的1,2感官刺激特征是高血压和压力反射介导的心动过缓发作。这些症状干扰日常生活脊髓损伤患者。因此，为了建立有效的工具，用于在动物中与脊髓损伤和试验性治疗心血管变化的调查是很重要的。 研究在动物的心血管功能，一些技术已经被用于监测血压和心脏速率。中央心血管参数可以通过套管插入和遥测记录，而无创尾袖口可以用来测量外周的血压3。相对于其他方法，远程13655具有的主要优势在于它允许连续记录中自由活动动物和长期监测心血管功能4。在脊髓损伤的动物模型，实验刺激后的变化周血压可能并不大到足以被检测到。因此，合适的心脏监测技术应该被选择用于动物SCI患者。 在本研究中，无线电遥测系统导入后脊髓横断监测成年大鼠心血管功能。大鼠接受的同基因大鼠胚胎第14天（E14）脑干源性神经干细胞（BS-神经干细胞）移植物在病灶部位。大鼠损伤，无移植和天真，没有受伤大鼠作为对照。遥测的步骤包括发送器灭菌并注入（ 图1），记录基底心血管参数，结直肠扩张诱导的反应，并且发送器的清洗和存储。

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			<td colspan="2" height="21" style="height:21px;width:315px;">Table of Specific Materials/Equipment:</td>
			<td style="width:103px;"></td>
			<td style="width:164px;"></td>
		</tr>
		<tr height="20">
			<td height="41" rowspan="2" style="height:41px;width:120px;">Reagents</td>
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		<tr height="21">
		</tr>
		<tr height="62">
			<td height="83" rowspan="2" style="height:83px;width:120px;">Fibrinogen (rat)</td>
			<td rowspan="2" style="width:195px;">Sigma</td>
			<td rowspan="2" style="width:103px;">F6755-25MG</td>
			<td rowspan="2" style="width:164px;">2 hr at 37oC to dissovle</td>
		</tr>
		<tr height="21">
		</tr>
		<tr height="59">
			<td height="80" rowspan="2" style="height:80px;width:120px;">Thrombin (rat)</td>
			<td rowspan="2" style="width:195px;">Sigma</td>
			<td rowspan="2" style="width:103px;">T5772-100UN</td>
			<td rowspan="2" style="width:164px;">Dissovle in 10 mM CaCl2</td>
		</tr>
		<tr height="21">
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:120px;">1% Terg-A-Zyme</td>
			<td style="width:195px;">Sigma</td>
			<td style="width:103px;">Z273287</td>
			<td style="width:164px;">Enzymatic solution for telemeter cleaning</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:120px;">Fluorogold</td>
			<td style="width:195px;">Fluorochrome</td>
			<td style="width:103px;"></td>
			<td style="width:164px;">Dissovle in distilled water and avoid light</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:120px;">Telemeter&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; (PA-C40)</td>
			<td style="width:195px;">Data Sciences International</td>
			<td style="width:103px;"></td>
			<td style="width:164px;"></td>
		</tr>
		<tr height="105">
			<td height="105" style="height:105px;width:120px;">Telementric recording and analysis system</td>
			<td style="width:195px;">Data Sciences International</td>
			<td style="width:103px;"></td>
			<td style="width:164px;">Signal stimulator, Data Exchange Matrix, receivers, Ambient pressure reference monitor</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:120px;">Balloon-tipped catheter</td>
			<td style="width:195px;">Edward Lifesciences</td>
			<td style="width:103px;">111F7-P</td>
			<td style="width:164px;">For colorectal distension</td>
		</tr>
	</tbody>
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a radio-telemetric system to monitor cardiovascular function in rats with spinal cord transection and embryonic neural stem cell grafts

高胸或颈脊髓损伤（SCI），可导致心脑血管功能障碍。为了监测心血管参数，我们植入连接到一个无线发射到大鼠的经历了T4脊髓横断伴或不表达绿色荧光蛋白的胚胎脑干源性神经干细胞移植的股动脉导管。相对于其他方法，例如套管插入或尾套，遥测有利的是连续地监测血压和心脏速率在自由活动动物。它也能够长期的多个数据采集。在脊髓损伤大鼠，针对结直肠扩张奔放的状态，自主神经反射异常在基础的心血管数据被成功记录。此外，前和损伤后的心血管参数可在相同的大鼠进行比较，如果发射器是脊髓横断前注入。所描述的telemet的一个限制Ry的过程是，植入股动脉可影响血液供给到同侧后肢。

使用上述的遥测技术，我们成功地记录在脊髓损伤的动物的心血管参数。在动物脊髓损伤独自一人，地图是人力资源，而相比增加了天真的动物，与以前的报告9一致显著减少。在动物BS-NSC嫁接，MAP和HR走近天真的动物测量水平（ 图2）。在结直肠扩张，一只老鼠被视为dysreflexic如果伤害性刺激所产生的上升在MAP和人力资源3,8的下降。阵发性高血压和压力感受器介导的心...

Watch this Scientific Journal Video about A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts at JoVE.com

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

我们提出了一个协议，使用无线电遥测系统，记录在T4脊髓横断大鼠心血管参数八个星期后胚胎脑干神经干细胞移植到病变部位。遥测是一种先进的技术来准确地评估在有意识的自由移动脊髓损伤大鼠的心血管功能。

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 ...

a-radio-telemetric-system-to-monitor-cardiovascular-function-rats

Research

JoVE Journal

Medicine

10.4K 次观看.  Drexel University College of Medicine. 我们提出了一个协议，使用无线电遥测系统，记录在T4脊髓横断大鼠心血管参数八个星期后胚胎脑干神经干细胞移植到病变部位。遥测是一种先进的技术来准确地评估在有意识的自由移动脊髓损伤大鼠的心血管功能。 

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

Assessing Endothelial Vasodilator Function with the Endo-PAT 2000

The endothelium is a delicate monolayer of cells that lines all blood vessels, and which comprises the systemic and lymphatic capillaries. By virtue of the panoply of paracrine factors that it secretes, the endothelium regulates the contractile and proliferative state of the underlying vascular smooth muscle, as well as the interaction of the vessel wall with circulating blood elements. Because of its central role in mediating vessel tone and growth, its position as gateway to circulating immune cells, and its local regulation of hemostasis and coagulation, the the properly functioning endothelium is the key to cardiovascular health. Conversely, the earliest disorder in most vascular diseases is endothelial dysfunction.
In the arterial circulation, the healthy endothelium generally exerts a vasodilator influence on the vascular smooth muscle. There are a number of methods to assess endothelial vasodilator function. The Endo-PAT 2000 is a new device that is used to assess endothelial vasodilator function in a rapid and non-invasive fashion. Unlike the commonly used technique of duplex ultra-sonography to assess flow-mediated vasodilation, it is totally non-operator-dependent, and the equipment is an order of magnitude less expensive. The device records endothelium-mediated changes in the digital pulse waveform known as the PAT ( peripheral Arterial Tone) signal, measured with a pair of novel modified plethysmographic probes situated on the finger index of each hand. Endothelium-mediated changes in the PAT signal are elicited by creating a downstream hyperemic response. Hyperemia is induced by occluding blood flow through the brachial artery for 5 minutes using an inflatable cuff on one hand. The response to reactive hyperemia is calculated automatically by the system. A PAT ratio is created using the post and pre occlusion values. These values are normalized to measurements from the contra-lateral arm, which serves as control for non-endothelial dependent systemic effects. Most notably, this normalization controls for fluctuations in sympathetic nerve outflow that may induce changes in peripheral arterial tone that are superimposed on the hyperemic response.
In this video we demonstrate how to use the Endo-PAT 2000 to perform a clinically relevant assessment of endothelial vasodilator function.

A noninvasive procedure to assess endothelial function is demonstrated using the Endo-PAT 2000.

评估与远藤的PAT 2000内皮血管舒张功能

The endothelium is a delicate monolayer of cells that lines all blood vessels, and which comprises the systemic and lymphatic capillaries. By virtue of ...

科研

医学

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 ...

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 ...

Synergetic Use of Neural Precursor Cells and Self-assembling Peptides in Experimental Cervical Spinal Cord Injury

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. Clinically, more than 50% of SCI affect the cervical spine1. As a consequence of the primary injury, a cascade of secondary mechanisms including inflammation, apoptosis, and demyelination occur finally leading to tissue scarring and development of intramedullary cavities2,3. Both represent physical and chemical barriers to cell transplantation, integration, and regeneration. Therefore, shaping the inhibitory environment and bridging cavities to create a supportive milieu for cell transplantation and regeneration is a promising therapeutic target4. Here, a contusion/compression model of cervical SCI using an aneurysm clip is described. This model is more clinically relevant than other experimental models, since complete transection or ruptures of the cord are rare. Also in comparison to the weight drop model, which in particular damage the dorsum columns, circumferential compression of the spinal cord appears advantageous. Clip closing force and duration can be adjusted to achieve different injury severity. A ring spring facilitates precise calibration and constancy of clip force. Under physiological conditions, synthetic self-assembling peptides (SAP) self-assemble into nanofibers and thus, are appealing for application in SCI5. They can be injected directly into the lesion minimizing damage to the cord. SAPs are biocompatible structures erecting scaffolds to bridge intramedullary cavities and thus, equip the damaged cord for regenerative treatments. K2(QL)6K2 (QL6) is a novel SAP introduced by Dong et al.6 In comparison to other peptides, QL6 self-assembles into &#946;-sheets at neutral pH6.14 days after SCI, after the acute stage, SAPs are injected into the center of the lesion and neural precursor cells (NPC) are injected into adjacent dorsal columns. In order to support cell survival, transplantation is combined with continuous subdural administration of growth factors by osmotic micro pumps for 7 days.

Treating cervical spinal cord injury with both self-assembling peptides (SAP) and neural precursor cells (NPC), together with growth factors, is a promising approach to promote regeneration and recovery. A contusion/compression aneurysm clip rat model of cervical SCI and combined treatment involving SAP injection and NPC transplantation is established.

在实验颈脊髓损伤协同使用神经前体细胞，并自组装肽的

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. ...

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 ...

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.

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) ...

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 ...

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. Research models for bypass graft failure are essential for a better understanding of pathobiological and pathophysiological processes during graft patency loss. Large animal models, such as pigs or sheep, resemble human anatomical structures but require special facilities and equipment. This video describes a rat vein interposition model to investigate vein graft patency loss. Rats are inexpensive and easy to handle. Compared to mouse models, the convenient size of rats permits better operability and enables a sufficient amount of material to be obtained for further diverse analysis. In brief, the inferior epigastric vein of a donor rat is harvested and used to replace a segment of the femoral artery. Anastomosis is conducted via single stitches and sealed with fibrin glue. Graft patency can be monitored non-invasively using duplex sonography. Myointimal hyperplasia, which is the main cause for graft patency loss, develops progressively over time and can be calculated from histological cross sections.

This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats.

静脉介入型号：研究旁路移植通畅一个合适的模型

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

Studying the Hypothalamic Insulin Signal to Peripheral Glucose Intolerance with a Continuous Drug Infusion System into the Mouse Brain

Insulin regulates systematic metabolism in the hypothalamus and the peripheral insulin response. An inflammatory reaction in peripheral adipose tissues contributes to type 2 diabetes mellitus (T2DM) development and appetite regulation in the hypothalamus. Chemokine CCL5 and C-C chemokine receptor type 5 (CCR5) levels have been suggested to mediate arteriosclerosis and glucose intolerance in type 2 diabetes mellitus (T2DM). In addition, CCL5 plays a neuroendocrine role in the hypothalamus by regulating food intake and body temperature, thus, prompting us to investigate its function in hypothalamic insulin signaling and the regulation of peripheral glucose metabolism.
The micro-osmotic pump brain infusion system is a quick and precise way to manipulate CCL5 function and study its effect in the brain. It also provides a convenient alternative approach to generating a transgenic knockout animal. In this system, CCL5 signaling was blocked by intracerebroventricular (ICV) infusion of its antagonist, MetCCL5, using a micro-osmotic pump. The peripheral glucose metabolism and insulin responsiveness was detected by the Oral Glucose Tolerance Test (OGTT) and Insulin Tolerance Test (ITT). Insulin signaling activity was then analyzed by protein blot from tissue samples derived from the animals.
After 7-14 days of MetCCL5 infusion, the glucose metabolism and insulin responsiveness was impaired in mice, as seen in the results of the OGTT and ITT. The IRS-1 serine302 phosphorylation was increased and the Akt activity was reduced in mice hypothalamic neurons following CCL5 inhibition. Altogether, our data suggest that blocking CCL5 in the mouse brain increases the phosphorylation of IRS-1 S302 and interrupts hypothalamic insulin signaling, leading to a decrease in insulin function in peripheral tissues as well as the impairment of glucose metabolism.

This protocol studies the role of chemokine (C-C motif) ligand 5 (CCL5) in the hypothalamus by delivering an antagonist, MetCCL5, into the mouse brain using a micro-osmotic pump brain infusion system. This transient inhibition of CCL5 activity interrupted hypothalamic insulin signaling, leading to glucose intolerance and peripheral systemic insulin sensitivity.

持续药物输注系统对小鼠脑内下丘脑胰岛素信号的研究

Insulin regulates systematic metabolism in the hypothalamus and the peripheral insulin response. An inflammatory reaction in peripheral adipose tissues ...

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine balloon denudation model, which is comparable with established vessel injury models in large animals. The aorta denudation model with balloon catheters mimics the clinical setting and leads to comparable pathobiological and physiological changes. Briefly, after performing a horizontal incision in the aorta abdominalis, a balloon catheter will be inserted into the vessel, inflated, and introduced retrogradely. Inflation of the balloon will lead to intima injury and overdistension of the vessel. After removing the catheter, the aortic incision will be closed with single stiches. The model shown in this article is reproducible, easy to perform, and can be established quickly and reliably. It is especially suitable for evaluating expensive experimental therapeutic agents, which can be applied in an economical fashion. By using different knockout-mouse strains, the impact of different genes on MH development can be assessed.

This article demonstrates a murine model to study the development of myointimal hyperplasia (MH) after aortic balloon injury.

球囊损伤诱发小鼠腹主动脉 Myointimal 增生的研究

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine ...