Name: urokinase-type plasminogen activator-induced mouse back pain model
Uploaded: 2023-09-01T12:00:00
Description: Watch this Scientific Journal Video about Urokinase-type Plasminogen Activator-induced Mouse Back Pain Model at JoVE.com

赠款资金由 NIH HEAL UG3 NS123958 提供。住房设施已通过AAALAC的检查和认可。动物被安置在动物资源中心（ARC）的住房设施中，由实验室工作人员和实验室和动物资源部（DLAR）工作人员维护。行为测试程序是美国疼痛协会和国际疼痛研究协会批准的该领域的标准方法。安乐死的方法符合美国兽医协会安乐死小组的建议。

所描述的所有动物程序均符合美国国立卫生研究院（NIH）的实验动物护理和使用指南。研究已获得新墨西哥大学健康科学中心当地机构护理和使用委员会 （IACUC #23-201364-HSC） 的批准。所有研究均符合OLAW合规保证（A3002-01）主持的关于在研究中使用动物的政策，如第三部分所述。II. 保证和认证。动物被安置在动物资源中心（ARC）的住房设施中，由实验室工作人员和实验室和动物资源部（DLAR）的工?...

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这种慢性背痛模型很容易诱发，在 1 周内建立的超敏反应可持续长达（甚至可能超过）8 周。这允许准确研究慢性疼痛状态，而不是其他仅持续一两周的急性模型。虽然我们在小鼠中展示了该模型，但 uPA 诱导的 CBP 模型也可以在大鼠2 中建立。该模型的一个优点是，长时间的疗程会引起焦虑和抑郁样行为的发展，这在慢性腰痛患者中观察到。鉴于背痛的普遍性16...

腰痛是最常见的残疾原因之一，全世界每 5 人中就有 1 人患有腰痛1。尽管做出了这些努力，但很少有可靠的背痛动物模型被广泛用于疼痛领域的动物研究，尤其是在小鼠中。以前的模型几乎完全利用大鼠来诱导慢性背痛 （CBP），例如通过将尿纤溶酶原激活剂 （uPA） 注射到腰椎小关节 2,3、将神经生长因子 （NGF） 注射到躯干肌肉组织4 或碘乙酸钠 （MIA）5 或白细胞介素-1β6 诱导的疼痛在椎间盘中注射。当然，大鼠是这些模型的首选，主要是因为它们的体型较大且易于注射炎症剂。
需要明确的是，背痛的小鼠模型确实存在，例如使用多年的椎间盘退行性SPARC无效小鼠模型7，但这些模型的建立比基于注射的模型更昂贵和耗时。最近的一项小鼠研究通过将 NGF 注射到具有垂直慢性约束压力的腰部肌肉中，建立了腰痛模型8。在以下方案中，我们已将来自大鼠的uPA诱导的CBP模型改编为小鼠2。超敏反应在 1 周内建立，并持续长达 6-8 周。此外，我们确定小鼠会发展出类似焦虑和抑郁的行为。鉴于背痛的普遍性和小鼠在分子疼痛研究中的更常见用途，这种持久的模型很容易被建立起来，用于开发缓解背痛的新治疗策略。

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			<td></td>
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			<td>70% ethanol</td>
			<td>Local Source</td>
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			<td>BALB/c mice</td>
			<td>Envigo</td>
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			<td>20-25 g</td>
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			<td>Cotton balls</td>
			<td>Fisher Scientific</td>
			<td>19-090-702</td>
			<td></td>
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			<td>Cotton-tipped applicators</td>
			<td>Fisher Scientific</td>
			<td>19-062-616</td>
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			<td>Isoflurane inhalant anesthetic</td>
			<td>MedVet</td>
			<td>RXISO-250</td>
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			<td>Labeling tape</td>
			<td>Fisher Scientific</td>
			<td>NGFP7002</td>
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			<td>Nitrile exam gloves</td>
			<td>Fisher Scientific</td>
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			<td>Local Source</td>
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			<td>Surgical drape, Steri-Drape Utility Sheet, Absorbent Prevention</td>
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			<td>76246-788</td>
			<td>cut into 15 x 15 cm pieces</td>
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			<td>Tygon tubing with 3 mm inner diameter</td>
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			<td>22XH87</td>
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			<td>Equipment</td>
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			<td>#11 carbon steel scalpel blades</td>
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			<td>Anesthesia induction chamber&#160;</td>
			<td>Summit Medical Equipment Company</td>
			<td>AS-01-0530-LG</td>
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			<td>Autoclave</td>
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			<td>Biology Dumont #5 forceps</td>
			<td>Fine Science Tools</td>
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			<td>Glass bead sterilizer Germinator 500</td>
			<td>VWR</td>
			<td>102095-946</td>
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			<td>IITC Life Sciences Series 8 Model PE34 Hot/Cold Plate Analgesia Meter</td>
			<td>IITC</td>
			<td>PE34</td>
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			<td>Integra Miltex cotton &#38; dressing pliers&#160;</td>
			<td>Safco Dental Supply</td>
			<td>66-317</td>
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			<td>OPTIKA CL31 double arm LED illuminator&#160;</td>
			<td>New York Microscope Company</td>
			<td>OPCL-31</td>
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			<td>Plantar Test System with InfraRed Emitter, i. e. Hargreaves Apparatus</td>
			<td>Ugo Basile</td>
			<td>37370-001 and 37370-002</td>
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			<td>Scalpel Handle No. 3</td>
			<td>VWR</td>
			<td>25607-947</td>
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			<td>Small animal heating pad&#160;</td>
			<td>Valley Vet Supply</td>
			<td>47375</td>
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			<td>Student Vannas spring scissors, straight blade&#160;</td>
			<td>Fine Science Tools</td>
			<td>91500-09</td>
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			<td>Table top animal research portable anesthesia workstation &#8220;PAM&#8221;</td>
			<td>Patterson Scientific</td>
			<td>AS-01-0007</td>
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			<td>Von Frey Filaments</td>
			<td>Ugo Basile</td>
			<td>37450-275</td>
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urokinase-type plasminogen activator-induced mouse back pain model

使用本文描述的简单方法可以在小鼠中诱导持续性腰痛的模型。这里提供了使用尿激酶型纤溶酶原激活剂（尿激酶）的简单、快速诱导小鼠持续性背痛模型的分步方法，尿激酶是一种存在于人类和其他动物中的丝氨酸蛋白酶。诱导小鼠持续性腰痛的方法包括沿腰椎韧带插入区域简单地注射尿激酶。尿激酶炎症因子将纤溶酶原激活为纤溶酶。通常，该模型可以在 10 分钟内诱导，并且超敏反应持续至少 8 周。
超敏反应、步态障碍和其他标准的焦虑和抑郁样测量可以在持续模型中进行测试。背痛是最普遍的疼痛类型。为了提高对背痛的认识，国际疼痛研究协会 （IASP） 将 2021 年命名为“全球背痛年”，将 2022 年命名为“将疼痛知识转化为实践的全球年”。疼痛疗法治疗进步的一个局限性是缺乏合适的模型来测试持续性和慢性疼痛。该模型的特点适用于测试旨在减轻背痛及其辅助特征的潜在疗法，有助于 IASP 将 2022 年命名为将疼痛知识转化为实践的全球年。

伤害性相关行为测试和数据分析 诱发措施 脚垫上的超敏反应在尿激酶注射后一天内发生。1周内，戒断阈值显著降低，持续到安乐死;这在术后第 4 周显示（图 4A）。使用 von Frey 上下方法9 和 Hargreaves 检验分析爪子退出潜伏期。在绘制的示例中，与对照组（n = 2 雄性，2 雌性）相比，具有 CBP 的小鼠（n = 4 雄性，4 雌性）表现出?...

Watch this Scientific Journal Video about Urokinase-type Plasminogen Activator-induced Mouse Back Pain Model at JoVE.com

Urokinase-type Plasminogen Activator-induced Mouse Back Pain Model

这里提供了使用韧带内注射尿纤溶酶原激活剂在小鼠中简单、快速诱导背痛模型的方法。

尿激酶型纤溶酶原激活剂诱导的小鼠背痛模型

A model of persisting lower back pain can be induced in mice with the simple methodology described herein. Step-by-step methods for simple, rapid ...

该协议概述了一种非侵入性方法，可在小鼠中诱导长达10周的慢性背痛，只需最少的训练即可快速可靠地进行。这种技术最困难的方面是识别注射韧带与注射肌肉的感觉。接触时感觉阻力表明韧带注射。没有阻力意味着针头可能在肌肉或腹腔中。首先，取一个无菌的平面放置鼠标。然后，在手术装置旁边放置一个加热垫恢复站，以在注射后立即转移小鼠。接下来，准备尿激酶和汉密尔顿注射器。事先绘制溶液，以尽量减少小鼠在麻醉下花费的时间。麻醉小鼠后，用两根手指定位肋骨下方L2和L3处的腰椎节段，并验证注射部位。将汉密尔顿注射器的尖端放在脊柱旁边。将注射器以大约 45 度角放置在紧邻骨骼的韧带中。轻轻但牢固地将针尖插入韧带，然后缓慢注射，直到注射完所有 5 微升。轻轻缓慢地取出针头。然后，将鼠标放入热回收站中，直到它醒来并变得移动。手术后检查小鼠一小时，以确保所有正常的运动功能继续。手术后，每天检查小鼠的体重和注射部位，持续一周，以防止任何感染或并发症。在测试之前，将每只动物移动到测试台上带有屏幕顶部的单独透明隔间中。然后，用 3.61 Von Frey 细丝刺激脚垫，该细丝可产生 4 克的力。足部退缩三到五次刺激被认为是积极的反应。现在，应用该系列中下一个较弱的细丝，直到动物停止对机械刺激做出反应。使用较高的灯丝，直到引起反应，然后切换回较低的灯丝。使用曲线拟合算法，计算机械撤回阈值，即在 50% 的时间内引起响应所需的最小力。要进行Hargreaves测试，请将小鼠放在由红外发射器从下方加热的玻璃表面上的隔间中。测量从对小鼠后爪施加红外光刺激到退出作为脚潜伏期的时间。对于冷板测试，将小鼠放在冷却至零下9摄氏度的冷板设备上。记录鼠标在放置在设备上后抬起脚所需的时间，作为撤回脚的延迟。要进行焦虑测试，请将每只动物放入位置偏好测试箱中，并在两个腔室之间留有通道。一个器械室应明亮，而另一侧应保持黑暗。在10分钟的测试中，使用计算机监控动物的明暗占用时间和过渡。接下来，将模型啮齿动物或幼稚动物放在零迷宫上，或使用两室明暗场所偏好测试。使用秒表评估在迷宫的封闭和开放区域花费的时间。对于蔗糖飞溅抑郁测试，在背侧喷洒 10% 至 30% 的蔗糖溶液，并对梳理的频率、持续时间和潜伏期进行评分，持续 10 分钟。为了进行新的物体测试，将小鼠单独安置在一个带有敞开顶部的透明塑料笼子中一小时。在笼子的相对角上添加两个相同的玩具迷你人偶，持续五分钟。在测试当天让动物适应一小时，并将两个相同的迷你人偶放在笼子的相同位置五分钟，然后将它们放回家中笼子。然后，用一个明显不同的新奇对象替换其中一个原始人物。四小时后，将小鼠放回测试笼中，并记录探索物体所花费的时间。对于运动功能评估，用在一侧纵向切割的纸巾管构建一个隧道。在一张干净的打印纸上铺开隧道。轻轻地握住老鼠，直到它们平静下来。将无毒的印度墨水涂在老鼠的爪子上，方法是将它们放入墨水池、印章垫或使用棉签。在隧道入口处释放老鼠。让他们穿过隧道并在最后抓住他们。根据步幅、步幅和脚趾张开对爪印进行评分。机械和热超敏反应试验表明，与幼稚对照组相比，慢性背痛小鼠的机械、热和冷敏感性显着增加。焦虑分析显示，慢性背痛小鼠在光暗盒光室中花费的总时间和饲养事件次数明显降低，表明焦虑。在蔗糖飞溅试验期间，患有慢性背痛的小鼠表现出较低的梳理时间和梳理次数。然而，梳理开始前的时间显着增加。患有慢性背痛的小鼠和幼稚的小鼠之间的步幅显着不同。尝试此程序时要记住的最重要的事情是，您的目标是脊柱周围的韧带，而不是肌肉。慢性背痛模型为评估各种新研究、药理学和其他潜在疗法提供了可靠的工具。

urokinase-type-plasminogen-activator-induced-mouse-back-pain-model

Research

JoVE Journal

Behavior

Assessing Functional Performance in the Mdx Mouse Model

Duchenne muscular dystrophy (DMD) is a severe and progressive muscle wasting disorder for which no cure is available. Nevertheless, several potential pharmaceutical compounds and gene therapy approaches have progressed into clinical trials. With improvement in muscle function being the most important end point in these trials, a lot of emphasis has been placed on setting up reliable, reproducible, and easy to perform functional tests to pre clinically assess muscle function, strength, condition, and coordination in the mdx mouse model for DMD. Both invasive and noninvasive tests are available. Tests that do not exacerbate the disease can be used to determine the natural history of the disease and the effects of therapeutic interventions (e.g. forelimb grip strength test, two different hanging tests using either a wire or a grid and rotarod running). Alternatively, forced treadmill running can be used to enhance disease progression and/or assess protective effects of therapeutic interventions on disease pathology. We here describe how to perform these most commonly used functional tests in a reliable and reproducible manner. Using these protocols based on standard operating procedures enables comparison of data between different laboratories.

Assessing Functional Performance in the Mdx Mouse Model

The primary outcome measure in clinical trials for neuromuscular disorders is generally improved muscle function. Therefore, assessing the effect of potential therapeutic compounds on muscle performance pre clinically in mouse models is of great importance. We here describe several functional tests to address this.

评估在功能性能<em&gt; MDX</em&gt;小鼠模型

Duchenne muscular dystrophy (DMD) is a severe and progressive muscle wasting disorder for which no cure is available. Nevertheless, several potential ...

科研

行为学

Determining Pain Detection and Tolerance Thresholds Using an Integrated, Multi-Modal Pain Task Battery

Human pain models are useful in the assessing the analgesic effect of drugs, providing information about a drug&#39;s pharmacology and identify potentially suitable therapeutic populations. The need to use a comprehensive battery of pain models is highlighted by studies whereby only a single pain model, thought to relate to the clinical situation, demonstrates lack of efficacy. No single experimental model can mimic the complex nature of clinical pain. The integrated, multi-modal pain task&#160;battery presented here encompasses the electrical stimulation task, pressure stimulation task, cold pressor task, the UVB inflammatory model which includes a thermal task and a paradigm for inhibitory conditioned pain modulation. These human pain models have been tested for predicative validity and reliability both in their own right and in combination, and can be used repeatedly, quickly, in short succession, with minimum burden for the subject and with a modest quantity of equipment. This allows a drug to be fully characterized and profiled for analgesic effect which is especially useful for drugs with a novel or untested mechanism of action.

Human pain models are valuable tools used to assess the analgesic potential of novel compounds and predict their clinical efficacy, especially when used in an integrated manner. Although implementation of these models is complex, with proper execution, the pain models described in this protocol can provide predictive and reliable results.

确定疼痛的检测和容忍阈值使用集成的，多模式疼痛任务电池

Human pain models are useful in the assessing the analgesic effect of drugs, providing information about a drug&#39;s pharmacology and identify ...

Treating Low Back Pain in Failed Back Surgery Patients with Multicolumn-lead Spinal Cord Stimulation

Failed back surgery syndrome (FBSS) refers to persistent, chronic pain following spinal surgery. Spinal cord stimulation with dorsal epidural leads can be used to treat back and leg pain in FBSS patients. This paper presents a detailed protocol for using spinal cord stimulation with surgical leads in FBSS patients. In our department, with the patient under general anesthesia, we place the lead in the epidural space by means of a small laminectomy at the 10th thoracic level. Placement of the lead is followed by a 1 month trial period with an externalized lead. If pain relief is greater than 50% at the end of this 1 month stimulation trial (required by Belgian reimbursement criteria), an internal pulse generator is then placed under the skin and connected to the lead in a second surgical procedure. We have demonstrated that using this technique in rigorously selected FBSS patients can significantly improve back pain, leg pain, patient activity, and quality of life for a sustained period of time.

This paper presents a method that uses spinal cord stimulation with a multicolumn lead to treat neuropathic low back pain in failed back surgery patients.

多列铅脊髓刺激术后腰痛的治疗

Failed back surgery syndrome (FBSS) refers to persistent, chronic pain following spinal surgery. Spinal cord stimulation with dorsal epidural leads can be ...

Measuring Active and Passive Tameness Separately in Mice

Domesticated animals such as dogs and laboratory mice show a high level of tameness, which is important for humans to handle them easily. Tameness has two behavioral components: a reluctance to avoid humans (passive tameness) and a motivation to approach humans (active tameness). To quantify these components in mice, we previously developed behavioral tests for active tameness, passive tameness, and the willingness to stay on a human hand, each designed to be completed within 3 min. The data obtained were used for selective breeding, with a large number of mice analyzed per generation. The active tameness test measures the movement of the mouse toward a human hand and the contact it engages in. The passive tameness test measures the duration of time that a mouse tolerates human touch. In the stay-on-hand test, a mouse is placed on a human hand and touched slowly using the thumb of that hand; the duration of time that the animal remains on the hand is measured. Here, we describe the test set-up and apparatus, explain the procedures, and discuss the appropriate data analysis. Finally, we explain how to interpret the results.

Tameness in animals includes a reduction of avoidance responses to humans (passive tameness) and an increase in active approaches to humans (active tameness). Here, we describe detailed protocols for three behavioral tests (active tameness, passive tameness, and stay-on-hand tests) to separately measure the active and passive tameness of mice.

小鼠主动和被动驯服的测定

Domesticated animals such as dogs and laboratory mice show a high level of tameness, which is important for humans to handle them easily. Tameness has two ...

Chronic Post-Ischemia Pain Model for Complex Regional Pain Syndrome Type-I in Rats

Complex regional pain syndrome type-I (CRPS-I) is a neurological disease that causes severe pain among patients and remains an unresolved medical condition. However, the underlying mechanisms of CRPS-I have yet to be revealed. It is known that ischemia/reperfusion is one of the leading factors that causes CRPS-I. By means of prolonged ischemia and reperfusion of the hind limb, the rat chronic post-ischemia pain (CPIP) model has been established to mimic CRPS-I. The CPIP model has become a well-recognized animal model for studying the mechanisms of CRPS-I. This protocol describes the detailed procedures involved in the establishment of the rat model of CPIP, including anesthesia, followed by ischemia/reperfusion of the hind limb. Characteristics of the rat CPIP model are further evaluated by measuring the mechanical and thermal hypersensitivities of the hind limb as well as the nocifensive responses to acute capsaicin injection. The rat CPIP model exhibits several CRPS-I-like manifestations, including hind limb edema and hyperemia in the early stage after establishment, persistent thermal and mechanical hypersensitivities, and increased nocifensive responses to acute capsaicin injection. These characteristics render it a suitable animal model for further investigation of the mechanisms involved in CRPS-I.

Provided here is a protocol that details steps to establish an animal model of chronic post-ischemia pain (CPIP). This is a well-recognized model mimicking human complex regional pain syndrome type-I. Mechanical and thermal hypersensitivities are further evaluated, as well as capsaicin-induced nocifensive behaviors observed in the CPIP rat model.

大鼠复杂区域疼痛综合征I型慢性缺血后疼痛模型

Complex regional pain syndrome type-I (CRPS-I) is a neurological disease that causes severe pain among patients and remains an unresolved medical ...

Investigating Pain-Related Avoidance Behavior using a Robotic Arm-Reaching Paradigm

Avoidance behavior is a key contributor to the transition from acute pain to chronic pain disability. Yet, there has been a lack of ecologically valid paradigms to experimentally investigate pain-related avoidance. To fill this gap, we developed a paradigm (the robotic arm-reaching paradigm) to investigate the mechanisms underlying the development of pain-related avoidance behavior. Existing avoidance paradigms (mostly in the context of anxiety research) have often operationalized avoidance as an experimenter-instructed, low-cost response, superimposed on stimuli associated with threat during a Pavlovian fear conditioning procedure. In contrast, the current method offers increased ecological validity in terms of instrumental learning (acquisition) of avoidance, and by adding a cost to the avoidance response. In the paradigm, participants perform arm-reaching movements from a starting point to a target using a robotic arm, and freely choose between three different movement trajectories to do so. The movement trajectories differ in probability of being paired with a painful electrocutaneous stimulus, and in required effort in terms of deviation and resistance. Specifically, the painful stimulus can be (partly) avoided at the cost of performing movements requiring increased effort. Avoidance behavior is operationalized as the maximal deviation from the shortest trajectory on each trial. In addition to explaining how the new paradigm can help understand the acquisition of avoidance, we describe adaptations of the robotic arm-reaching paradigm for (1) examining the spread of avoidance to other stimuli (generalization), (2) modeling clinical treatment in the lab (extinction of avoidance using response prevention), as well as (3) modeling relapse, and return of avoidance following extinction (spontaneous recovery). Given the increased ecological validity, and numerous possibilities for extensions and/or adaptations, the robotic arm-reaching paradigm offers a promising tool to facilitate the investigation of avoidance behavior and to further our understanding of its underlying processes.

Avoidance is central to chronic pain disability, yet adequate paradigms for examining pain-related avoidance are lacking. Therefore, we developed a paradigm that allows investigating how pain-related avoidance behavior is learned (acquisition), spreads to other stimuli (generalization), can be mitigated (extinction), and how it may subsequently re-emerge (spontaneous recovery).

使用机械臂接触范式调查疼痛相关避免行为

Avoidance behavior is a key contributor to the transition from acute pain to chronic pain disability. Yet, there has been a lack of ecologically valid ...

Back Mechanical Sensitivity Assessment in the Rat for Mechanistic Investigation of Chronic Back Pain

Low back pain is the leading cause of disability worldwide, with dramatic personal, economic, and social consequences. To develop novel therapeutics, animal models are needed to examine the mechanisms and effectiveness of novel therapies from a translational perspective. Several rodent models of back pain are used in current investigations. Surprisingly, however, no standardized behavioral test was validated to assess mechanical sensitivity in back pain models. This is critical to confirm that animals with presumed back pain present local hypersensitivity to nociceptive stimuli, and to monitor sensitivity during interventions designed to relieve back pain. The objective of this study is to lay down a simple and accessible test to assess mechanical sensitivity in the back of rats. A test cage was fabricated specifically for this method; length x width x height: 50 x 20 x 7 cm, having a stainless-steel mesh on the top. This test cage allows the application of mechanical stimuli to the back. To perform the test, the back of the animal is shaved in the region of interest, and the test area is marked to repeat the test on different days, as needed. The mechanical threshold is determined with Von Frey filaments applied to the paraspinal muscles, utilizing the up-down method described previously. The positive responses include (1) muscle twitching, (2) arching (back extension), (3) rotation of the neck (4) scratching or licking the back, and (5) escaping. This behavioral test (Back Mechanical Sensitivity (BMS) test) is useful for mechanistic research with rodent models of back pain for the development of therapeutic interventions for the prevention and management of back pain.

To develop novel therapeutic interventions for the prevention and management of back pain, animal models are required to examine the mechanisms and effectiveness of these therapies from a translational perspective. The present protocol describes the BMS test, a standardized method to assess back mechanical sensitivity in the rat.

大鼠背部机械敏感性评估用于慢性背痛的机制研究

Low back pain is the leading cause of disability worldwide, with dramatic personal, economic, and social consequences. To develop novel therapeutics, ...

Elevated Plus Maze Test in an Angelman Syndrome Mouse Model

To begin, place the plus shaped maze on the testing platform just below the camera. Using the potentiometer on the wall, adjust the light intensity to 70 luxe at its center with the help of a luxometer with its sensor placed at the center of the maze during the adjustment. Open the software by double clicking on the viewer software icon and load the configuration for Elevated Plus Maze or EPM testing by clicking on the icon on the upper left side of the configuration tab.Enter the animal information into the corresponding fields of the experiment tab including animal id, genotype, gender, and experiment information. Verify that the zone's position, open arms, and closed arms are correctly configured. Using a visual control and computer mouse, ensure that the virtual outline zones match the corresponding EPM zones.Place the mouse cursor on the arrow icon on the upper left side of the acquisition tab. Remove the animal from the home cage and place it gently in the center of the EPM. Start the experiment by left clicking on the computer mouse.After five minutes, save the recorded data by clicking okay. Name the file and click save. Export the results to a csv file for offline analysis by clicking on the icon on the left vertical panel of the data analysis tab.In the EPM test, none of the parameters related to anxiety-like behavior were altered in maternal heterozygous Ube3A mutants.

天使综合征小鼠模型中的升高加迷宫试验

Open Field Test in an Angelman Syndrome Mouse Model

Place the four open field, or OF, test boxes on the testing platform below the camera. Using the potentiometer on the wall, adjust the light intensity to 200 lux at the center of each OF test with the help of a luxometer with its sensor placed at the center of each box during the adjustment. Open the software by double clicking on the viewer software icon and load the configuration for EF testing by clicking on the icon on the upper left side of the configuration tab.Enter the animal information into the corresponding fields of the experiment tab. Verify that the zone's position matches the OF test boxes and adjust them if needed. Using a visual control and computer mouse, ensure that the virtual outline center and periphery zones match the corresponding OF test zones.Place the mouse cursor on the arrow icon on the upper left side of the acquisition tab. Remove four animals from the home cage and place them in the corner of each test box. Stop the experiment by left clicking on the computer mouse.In the OF test, the maternal heterozygous Ube3a mutant animals were significantly hypoactive as reflected by a shorter traversed distance, lower average speed, and longer resting time. However, none of the parameters related to anxiety-like behavior were altered in Ube3a mutants.

安格曼综合征小鼠模型中的开放现场测试

Tail Suspension Test in an Angelman Syndrome Mouse Model

Connect the tail suspension cage to the computer using a USB cable. Insert the USB dongle into the computer and start the software by double clicking on the BioTSD software icon. In the settings tab on the global, ensure the acquisition duration is set to 360 seconds.In the experiment tab, select the new list of subjects and create a new experiment file and a new list of tested subjects by following the instructions in the opened tab. Prepare the animals for the test by wrapping single-sided adhesive tape around three-fourths of the animal's tail starting from the base. Start the run by clicking start run then continue in the acquisition tab.Immediately after hanging the animal on the hook, click the start icon under the visualized position to acquire data. After testing all the animals, in the analysis tab, select the last four minutes of the acquisition for analysis. Select all valid runs in the analysis period and click analyze selected subjects.Choose the desired data format and click export selected data to export the collected data. In the tail suspension test, Ube3a mutant animals were immobile for significantly longer than their control littermates, indicating their depression-like behavior.