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。

<table><tbody><tr><td>Fibrinogen (rat)</td><td>Sigma</td><td>F6755-25MG</td><td>2 hr at 37 &lt;sup&gt;o&lt;/sup&gt;C to dissovle</td></tr><tr><td>Thrombin (rat)</td><td>Sigma</td><td>T5772-100UN</td><td>Dissovle in 10 mM CaCl2</td></tr><tr><td>1% Terg-A-Zyme</td><td>Sigma</td><td>Z273287</td><td>Enzymatic solution for telemeter cleaning</td></tr><tr><td>Fluorogold</td><td>Fluorochrome</td><td></td><td>Dissovle in distilled water and avoid light</td></tr><tr><td>Telemeter&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; (PA-C40)</td><td>Data Sciences International</td><td></td><td></td></tr><tr><td>Telementric recording and analysis system</td><td>Data Sciences International</td><td></td><td>Signal stimulator, Data Exchange Matrix, receivers, Ambient pressure reference monitor</td></tr><tr><td>Balloon-tipped catheter</td><td>Edward Lifesciences</td><td>111F7-P</td><td>For colorectal distension</td></tr></tbody></table>

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

Universidad de Cartagena UDEC

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

Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice

The laboratory mouse is the animal species of choice for most biomedical research, in both the academic sphere and the pharmaceutical industry. Mice are a manageable size and relatively easy to house. These factors, together with the availability of a wealth of spontaneous and experimentally induced mutants, make laboratory mice ideally suited to a wide variety of research areas.
In cardiovascular, pharmacological and toxicological research, accurate measurement of parameters relating to the circulatory system of laboratory animals is often required. Determination of heart rate, heart rate variability, and duration of PQ and QT intervals are based on electrocardiogram (ECG) recordings. However, obtaining reliable ECG curves as well as physiological data such as core body temperature in mice can be difficult using conventional measurement techniques, which require connecting sensors and lead wires to a restrained, tethered, or even anaesthetized animal. Data obtained in this fashion must be interpreted with caution, as it is well known that restraining and anesthesia can have a major artifactual influence on physiological parameters1, 2.
Radiotelemetry enables data to be collected from conscious and untethered animals. Measurements can be conducted even in freely moving animals, and without requiring the investigator to be in the proximity of the animal. Thus, known sources of artifacts are avoided, and accurate and reliable measurements are assured. This methodology also reduces interanimal variability, thus reducing the number of animals used, rendering this technology the most humane method of monitoring physiological parameters in laboratory animals3, 4. Constant advancements in data acquisition technology and implant miniaturization mean that it is now possible to record physiological parameters and locomotor activity continuously and in realtime over longer periods such as hours, days or even weeks3, 5.
Here, we describe a surgical technique for implantation of a commercially available telemetry transmitter used for continuous measurements of core body temperature, locomotor activity and biopotential (i.e. onelead ECG), from which heart rate, heart rate variability, and PQ and QT intervals can be established in freeroaming, untethered mice. We also present pre-operative procedures and protocols for post-operative intensive care and pain treatment that improve recovery, well-being and survival rates in implanted mice5, 6.

A surgical technique for implantation of commercially available telemetry transmitters used for continuous measurement of biopotential (one-lead ECG), heart rate, core body temperature and locomotor activity in freely moving mice is shown. Recommendations and protocols for post-operative care and pain relief, improving recovery, well being and survival rate are also presented.

植入的无线电遥测发射器产生心电图，心率，身体核心温度和自由移动的实验室小鼠的活动上的数据

The laboratory mouse is the animal species of choice for most biomedical research, in both the academic sphere and the pharmaceutical industry. Mice are a ...

科研

医学

Using Laser Doppler Imaging and Monitoring to Analyze Spinal Cord Microcirculation in Rat

Laser Doppler flowmetry (LDF) is a noninvasive method for blood flow (BF) measurement, which makes it preferable for measuring microcirculatory alterations of the spinal cord. In this article, our goal was to use both Laser Doppler imaging and monitoring to analyze the change of BF after spinal cord injury. Both the laser Doppler image scanner and the probe/monitor were being employed to obtain each readout. The data of LDPI provided a local distribution of BF, which gave an overview of perfusion around the injury site and made it accessible for comparative analysis of BF among different locations. By intensely measuring the probing area over a period of time, a combined probe was used to simultaneously measure the BF and oxygen saturation of the spinal cord, showing overall spinal cord perfusion and oxygen supply. LDF itself has a few limitations, such as relative flux, sensitivity to movement, and biological zero signal. However, the technology has been applied in clinical and experimental study due to its simple setup and rapid measurement of BF.

Here we present a combination of laser Doppler perfusion imaging (LDPI) and laser Doppler perfusion monitoring (LDPM) to measure spinal cord local blood flows and oxygen saturation (SO2), as well as a standardized procedure for introducing spinal cord trauma on rat.

激光多普勒成像与监测在大鼠脊髓微循环分析中的应用

Laser Doppler flowmetry (LDF) is a noninvasive method for blood flow (BF) measurement, which makes it preferable for measuring microcirculatory ...

行为学

Promotion of Survival and Differentiation of Neural Stem Cells with Fibrin and Growth Factor Cocktails after Severe Spinal Cord Injury

Neural stem cells (NSCs) can self-renew and differentiate into neurons and glia. Transplanted NSCs can replace lost neurons and glia after spinal cord injury (SCI), and can form functional relays to re-connect spinal cord segments above and below a lesion. Previous studies grafting neural stem cells have been limited by incomplete graft survival within the spinal cord lesion cavity. Further, tracking of graft cell survival, differentiation, and process extension had not been optimized. Finally, in previous studies, cultured rat NSCs were typically reported to differentiate into glia when grafted to the injured spinal cord, rather than neurons, unless fate was driven to a specific cell type. To address these issues, we developed new methods to improve the survival, integration and differentiation of NSCs to sites of even severe SCI. NSCs were freshly isolated from embryonic day 14 spinal cord (E14) from a stable transgenic Fischer 344 rat line expressing green fluorescent protein (GFP) and were embedded into a fibrin matrix containing growth factors; this formulation aimed to retain grafted cells in the lesion cavity and support cell survival. NSCs in the fibrin/growth factor cocktail were implanted two weeks after thoracic level-3 (T3) complete spinal cord transections, thereby avoiding peak periods of inflammation. Resulting grafts completely filled the lesion cavity and differentiated into both neurons, which extended axons into the host spinal cord over remarkably long distances, and glia. Grafts of cultured human NSCs expressing GFP resulted in similar findings. Thus, methods are defined for improving neural stem cell grafting, survival and analysis of in vivo findings.

Fibrin matrices containing growth factors were used to retain grafted neural stem cells into sites of complete spinal cord transection. Grafted cells completely filled the lesion cavity and differentiated into multiple neural cell types, including neurons that extended axons into host spinal cord over long distances.

促进存活神经干细胞与纤维蛋白和生长因子鸡尾酒和分化的严重脊髓损伤后

Neural stem cells (NSCs) can self-renew and differentiate into neurons and glia. Transplanted NSCs can replace lost neurons and glia after spinal cord ...

神经科学

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 ...

Long-term Continuous EEG Monitoring in Small Rodent Models of Human Disease Using the Epoch Wireless Transmitter System

Many progressive neurologic diseases in humans, such as epilepsy, require pre-clinical animal models that slowly develop the disease in order to test interventions at various stages of the disease process. These animal models are particularly difficult to implement in immature rodents, a classic model organism for laboratory study of these disorders. Recording continuous EEG in young animal models of seizures and other neurological disorders presents a technical challenge due to the small physical size of young rodents and their dependence on the dam prior to weaning. Therefore, there is not only a clear need for improving pre-clinical research that will better identify those therapies suitable for translation to the clinic but also a need for new devices capable of recording continuous EEG in immature rodents. Here, we describe the technology behind and demonstrate the use of a novel miniature telemetry system, specifically engineered for use in immature rats or mice, which is also effective for use in adult animals.

Here we demonstrate the use of a wireless enabling technology for electroencephalogram (EEG) in neonatal rodent models of human disease. With telemetry, there are no encumbering connections, thus allowing natural behaviors.

在人类疾病中使用大纪元无线传输系统的小型啮齿动物模型的长期连续监测脑电图

Many progressive neurologic diseases in humans, such as epilepsy, require pre-clinical animal models that slowly develop the disease in order to test ...

Development of an Algorithm to Perform a Comprehensive Study of Autonomic Dysreflexia in Animals with High Spinal Cord Injury Using a Telemetry Device

Spinal cord injury (SCI) is a debilitating neurological condition characterized by somatic and autonomic dysfunctions. In particular, SCI above the mid-thoracic level can lead to a potentially life-threatening hypertensive condition called autonomic dysreflexia (AD) that is often triggered by noxious or non-noxious somatic or visceral stimuli below the level of injury. One of the most common triggers of AD is the distension of pelvic viscera, such as during bladder and bowel distension or evacuation. This protocol presents a novel pattern recognition algorithm developed for a JAVA platform software to study the fluctuations of cardiovascular parameters as well as the number, severity and duration of spontaneously occurring AD events. The software is able to apply a pattern recognition algorithm on hemodynamic data such as systolic blood pressure (SBP) and heart rate (HR) extracted from telemetry recordings of conscious and unrestrained animals before and after thoracic (T3) complete transection. With this software, hemodynamic parameters and episodes of AD are able to be detected and analyzed with minimal experimenter bias.

The catheter of a telemetry device is implanted into the abdominal aorta in order to continuously collect beat-by-beat hemodynamic data from animals pre and post-high thoracic spinal cord transection. A novel JAVA software was employed to analyze hemodynamic parameters as well as frequency and intensity of spontaneous episodes of autonomic dysreflexia.

一个算法的开发与高位脊髓损伤使用遥测装置进行自主神经异常反射的动物的综合研究

Spinal cord injury (SCI) is a debilitating neurological condition characterized by somatic and autonomic dysfunctions. In particular, SCI above the ...

Wireless Intramuscular EMG System for Locomotion Studies in Stroke Model Rats

A Real-Time Wearable Electromyography Measurement System for Small Animals

The intramuscular electromyography (EMG) measurement method for experimental animals has been implemented in various ways. Among these methods, tethering cables to external measurement devices can restrict the movement of experimental animals, while implantable devices may cause unwanted side effects due to the constant presence of a device with considerable size and weight. To address these issues, we propose a low-cost, wireless, detachable EMG measurement system and experimental procedure. This article focuses on the surgical installation of intramuscular wire electrodes with small connectors and the development of the wireless system. Notably, in this system, only the wire electrodes are inserted into the animal's body. Using this system, EMG measurements can be easily performed by attaching the circuit system to a connector installed on the animal's back, with real-time monitoring achievable on a laptop. The proposed method is explained in a detailed, step-by-step manner, followed by a demonstration involving the insertion of intramuscular electrodes into the hindlimbs of a rat. A treadmill experiment is conducted for a locomotion study, and the resulting electrophysiological signals are subsequently obtained and analyzed.

A protocol is presented for the installation and preparation of intramuscular electrodes, along with an animal-mountable, miniaturized measurement device for electromyography (EMG) analysis in locomotion studies. This system enables the wireless transmission of real-time EMG data for investigating locomotion recovery in stroke model rats.

The intramuscular electromyography (EMG) measurement method for experimental animals has been implemented in various ways. Among these methods, tethering ...

工程学

Novel Approach for Simultaneous Recording of Renal Sympathetic Nerve Activity and Blood Pressure with Intravenous Infusion in Conscious, Unrestrained Mice.

Renal sympathetic nerves contribute significantly to both physiological and pathophysiological phenomena. Evaluating renal sympathetic nerve activity (RSNA) is of great interest in many areas of research such as chronic kidney disease, hypertension, heart failure, diabetes and obesity. Unequivocal assessment of the role of the sympathetic nervous system is thus imperative for proper interpretation of experimental results and understanding of disease processes. RSNA has been traditionally measured in anesthetized rodents, including mice. However, mice usually exhibit very low systemic blood pressure and hemodynamic instability for several hours during anesthesia and surgery. Meaningful interpretation of RSNA is confounded by this non-physiological state, given the intimate relationship between sympathetic nervous tone and cardiovascular status. To address this limitation of traditional approaches, we developed a new method for measuring RSNA in conscious, freely-moving mice. Mice were chronically instrumented with radio-telemeters for continuous monitoring of blood pressure as well as a jugular venous infusion catheter and custom-designed bipolar electrode for direct recording of RSNA. Following a 48-72 hour recovery period, survival rate was 100% and all mice behaved normally. At this time-point, RSNA was successfully recorded in 80% of mice, with viable signals acquired up to 4 and 5 days post-surgery in 70% and 50% of mice, respectively. Physiological blood pressures were recorded in all mice (116&plusmn;2 mmHg; n=10). Recorded RSNA increased with eating and grooming, as well-established in the literature. Furthermore, RSNA was validated by ganglionic blockade and modulation of blood pressure with pharmacological agents. Herein, an effective and manageable method for clear recording of RSNA in conscious, freely-moving mice is described.

Anesthetized mice exhibit non-physiological systemic blood pressure, which precludes meaningful assessment of autonomic tone given the intimate relationship between blood pressure and the autonomic nervous system. Thus, a novel method to simultaneously record renal sympathetic nerve activity and blood pressure with intravenous infusion in conscious mice is outlined.

清醒、无拘无束的小鼠静脉输液同时记录肾交感神经活动和血压的新方法。

Renal sympathetic nerves contribute significantly to both physiological and pathophysiological phenomena. Evaluating renal sympathetic nerve activity ...

Transplantation of Schwann Cells Inside PVDF-TrFE Conduits to Bridge Transected Rat Spinal Cord Stumps to Promote Axon Regeneration Across the Gap

Among various models for spinal cord injury in rats, the contusion model is the most often used because it is the most common type of human spinal cord injury. The complete transection model, although not as clinically relevant as the contusion model, is the most rigorous method to evaluate axon regeneration. In the contusion model, it is difficult to distinguish regenerated from sprouted or spared axons due to the presence of remaining tissue post injury. In the complete transection model, a bridging method is necessary to fill the gap and create continuity from the rostral to the caudal stumps in order to evaluate the effectiveness of the treatments. A reliable bridging surgery is essential to test outcome measures by reducing the variability due to the surgical method. The protocols described here are used to prepare Schwann cells (SCs) and conduits prior to transplantation, complete transection of the spinal cord at thoracic level 8 (T8), insert the conduit, and transplant SCs into the conduit. This approach also uses in situ gelling of an injectable basement membrane matrix with SC transplantation that allows improved axon growth across the rostral and caudal interfaces with the host tissue.

This article describes a technique to insert a hollow conduit between the spinal cord stumps after complete transection and fill with Schwann cells (SCs) and injectable basement membrane matrix in order to bridge and promote axon regeneration across the gap.

氟 TrFE 导管内许旺细胞移植桥断大鼠脊髓残肢促进轴突再生跨越间隙

Among various models for spinal cord injury in rats, the contusion model is the most often used because it is the most common type of human spinal cord ...

A Neurosphere Assay to Evaluate Endogenous Neural Stem Cell Activation in a Mouse Model of Minimal Spinal Cord Injury

Neural stem cells (NSCs) in the adult mammalian spinal cord are a relatively mitotically quiescent population of periventricular cells that can be studied in vitro using the neurosphere assay. This colony-forming assay is a powerful tool to study the response of NSCs to exogenous factors in a dish; however, this can also be used to study the effect of in vivo manipulations with the proper understanding of the strengths and limitations of the assay. One manipulation of the clinical interest is the effect of injury on endogenous NSC activation. Current models of spinal cord injury provide a challenge to study this as the severity of common contusion, compression, and transection models cause the destruction of the NSC niche at the site of the injury where the stem cells reside. Here, we describe a minimal injury model that causes localized damage at the superficial dorsolateral surface of the lower thoracic level (T7/8) of the adult mouse spinal cord. This injury model spares the central canal at the level of injury and permits analysis of the NSCs that reside at the level of the lesion at various time points following injury. Here, we show how the neurosphere assay can be utilized to study the activation of the two distinct, lineally-related, populations of NSCs that reside in the spinal cord periventricular region - primitive and definitive NSCs (pNSCs and dNSCs, respectively). We demonstrate how to isolate and culture these NSCs from the periventricular region at the level of injury and the white matter injury site. Our post-surgical spinal cord dissections show increased numbers of pNSC and dNSC-derived neurospheres from the periventricular region of injured cords compared to controls, speaking to their activation via injury. Furthermore, following injury, dNSC-derived neurospheres can be isolated from the injury site &#8212; demonstrating the ability of NSCs to migrate from their periventricular niche to sites of injury.

Here, we demonstrate the performance of a minimal spinal cord injury model in an adult mouse that spares the central canal niche housing endogenous neural stem cells (NSCs). We show how the neurosphere assay can be used to quantify activation and migration of definitive and primitive NSCs following injury.

小鼠脊髓损伤模型中内源性神经干细胞活化的干细胞检测

Neural stem cells (NSCs) in the adult mammalian spinal cord are a relatively mitotically quiescent population of periventricular cells that can be studied ...