Name: burn injury-induced pain and depression-like behavior in mice
Uploaded: 2021-09-29T14:00:00
Description: Watch this Scientific Journal Video about Burn Injury-Induced Pain and Depression-Like Behavior in Mice at JoVE.com

本研究得到了忠南国立大学和韩国政府资助的韩国国家研究基金会（NRF-2019R1A6A3A01093963和NRF-2021R1F1A1062509）的支持。

所有实验方案均由韩国忠南国立大学动物护理和使用委员会审查和批准，然后根据国际疼痛研究协会18的伦理准则进行。
1.后爪烫伤的诱导
<ol><li>将体重为20-25g的雄性ICR小鼠置于光照和温控室（12/12小时光暗循环，22.5°C±2.5°C），湿度为40%-60%。 注意：雄性和雌性小鼠都可用于此方案。</li>
	<li>让动物自由接触食物和水，并在开始实验前适应环...

<ol>
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烫伤是一种由加热的液体引起的热灼伤。有人认为，在大多数情况下会发生一度或二度烧伤，但长期接触热源可导致三度烧伤26。在本研究中，通过将小鼠的右后爪暴露在65°C的热水中3秒4，26来诱导烧伤。在烧伤的爪子中检测到组织损伤，其显示烧伤的常见症状，例如发红，皮肤剥落和肿胀（补充图1）4?...

组织损伤，如烧伤和创伤，通常与急性疼痛同时发生有关。烧伤和创伤相关症状估计每年有1，80，000例死亡是由烧伤引起的 - 绝大多数发生在低收入和中等收入国家，来自不同类型烧伤1。根据一份全球报告，烧伤在儿童中很常见，约占住院患者的40%-60%2，3。这些特定的伤害甚至更严重，因为它们可能发生在日常生活中，例如煮沸或洗澡水4，5。虽然在大多数情况下，急性疼痛可以在从组织损伤中恢复后自发解决，但由于神经系统的异常变化6，7，它可能变成慢性疼痛。
最近，有人提出急性疼痛可以诱发抑郁情绪，慢性疼痛可以引起焦虑和抑郁8，9，10，11。疼痛和抑郁的共存使患者更难治疗。抑郁症也倾向于增加疼痛敏感性，这很可能诱发更强烈的抑郁症和疼痛12。疼痛和抑郁症的并发症显示在外周炎症的动物模型中13，14，15，16。疼痛诱发的抑郁症的详细机制直到现在还不是很清楚17.因此，有必要开发更有效的烧伤治疗方法，以减轻副作用和症状。
因此，本研究旨在开发一种动物模型来研究烧伤引起的急性疼痛和小鼠抑郁样行为。为此，测量了烧伤相关的异常触觉敏感性，步态模式改变和抑郁样行为。此外，本研究还尝试使用非甾体抗炎药验证模型。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1 mL syringe</td>
			<td>BD</td>
			<td>307809</td>
			<td></td>
		</tr>
		<tr>
			<td>1.5 mL tube</td>
			<td>Axygen</td>
			<td>MCT-150-C</td>
			<td></td>
		</tr>
		<tr>
			<td>50 mL tube</td>
			<td>SPL</td>
			<td>50050</td>
			<td></td>
		</tr>
		<tr>
			<td>Acetaminophen BioXtra, &#8805;99.0%</td>
			<td>Sigma-Aldrich</td>
			<td>A7085-100G</td>
			<td>Positive control (The analgesic agent, acetaminophen (200 mg/kg) was administered intraperitoneally once-daily for 7 days starting from the day of after burn injury (Only Burn + Acetaminophen group. (von-Frey test, gait analysis, and forced swimming test: Used for drug-dependent behavioral testing after burn injury), (Rota-rod test: It was used to investigate the motor and functional impairments of the drug in animals after burn injury).</td>
		</tr>
		<tr>
			<td>Alfaxan multidose (Alfaxalone)</td>
			<td>JUROX Pty.Limited</td>
			<td></td>
			<td>In this experiment, this material used for animal anesthesia, and was used as a positive control for experimentally treated drugs in the rota-rod test.</td>
		</tr>
		<tr>
			<td>CatWalk automated gait analysis system</td>
			<td>Noldus</td>
			<td>CatWalk XT</td>
			<td>Gait analysis in freely walking rodents is used to study the changes in limb movement and positioning in models with sensory-motor dysfunction</td>
		</tr>
		<tr>
			<td>OPTISHIELD (Cyclosporin ophthalmic ointment)</td>
			<td>Ashish Life Science</td>
			<td></td>
			<td>In this experiment, this material was used for an ointment to prevent corneal drying after induction of anesthesia.</td>
		</tr>
		<tr>
			<td>Plexiglass cylinder</td>
			<td>SCITECH KOREA</td>
			<td>custom made products</td>
			<td>Used in forced swimming test</td>
		</tr>
		<tr>
			<td>Rota-rod system</td>
			<td>SCITECH KOREA</td>
			<td>Accelerating rota rod</td>
			<td>Used in the measurement of Normal Motor Function</td>
		</tr>
		<tr>
			<td>von Frey filaments</td>
			<td>North Coast Medical</td>
			<td>NC12775</td>
			<td>Used in the measurement of Mechanical Allodynia</td>
		</tr>
		<tr>
			<td>Waterbath</td>
			<td>CHANGSHIN SCIENCE</td>
			<td>C-WBE</td>
			<td>Used in the burn injury induction</td>
		</tr>
	</tbody>
</table>

burn injury-induced pain and depression-like behavior in mice

烫水是老年人和年轻人烧伤的最常见原因。这是主要的临床挑战之一，因为低收入和中等收入国家的死亡率和后遗症很高。烧伤经常诱发强烈的自发性疼痛和持续性异常疼痛以及危及生命的问题。更重要的是，过度疼痛往往伴有抑郁症，这可能会显著降低生活质量。本文展示了如何开发一种动物模型来研究烧伤引起的疼痛和抑郁样行为。麻醉后，通过将小鼠的一只后爪浸入热水（65°C±0.5°C）中3s诱导烧伤。烧伤后每2天进行一次von Frey测试和自动步态分析。此外，使用强制游泳试验检查抑郁样行为，并进行旋转杆试验以区分烧伤后异常的运动功能。这项研究的主要目的是描述用于研究烧伤引起的疼痛和小鼠抑郁样行为的动物模型的发展。

为了最大限度地减少动物的痛苦并减少根据三个Rs（替换，减少和改进）指南使用的动物数量，本研究的设计采用了最小数量的动物，用于收集通过初步实验建立的重要数据。在这项研究中，行为实验独立进行了两次，如下所示。步态分析，机械性异常和抑郁样行为测试与对照组（n = 5），烧伤（n = 7;载体对照;盐水）和烧伤+对乙酰氨基酚（n = 7）组一起进行。在轮状杆试验中，对照（n = 3），燃烧...

Watch this Scientific Journal Video about Burn Injury-Induced Pain and Depression-Like Behavior in Mice at JoVE.com

Burn Injury-Induced Pain and Depression-Like Behavior in Mice

一个后爪的短暂性烫伤（65 °C ± 0.5 °C，3 s）降低了同侧冯弗雷丝刺激的阈值（g）并改变了步态模式。此外，烧伤在强制游泳测试中诱发抑郁样行为。

烧伤诱导的疼痛和小鼠抑郁样行为

Scalding water is the most common cause of burn injury in both elderly and young populations. It is one of the major clinical challenges because of the ...

该方案适用于烧伤疼痛及其治疗的各个方面和结果的研究，有望为该研究领域带来重要信息。该协议建立了热水诱导的烧伤动物模型，并提出了一种测量相关疼痛，步态障碍和抑郁样行为反应的方法。演示该程序的将是Kim博士实验室的硕士生Jaehyuk Kim。首先，将体重20至25克的雄性ICR小鼠安置在光照和温度受控的房间里。让小鼠在实验开始前自由获得食物和水，并适应环境至少一周。一周后，将小鼠随机分配到实验组或对照组，并使用动物编号作为进行盲实验的代码。在进行烧伤诱导之前，请穿戴手术服，手套和口罩。接下来，在深度麻醉小鼠后，用70%乙醇消毒右后爪周围，并在眼睛上涂抹眼药膏以防止角膜干燥，然后在每只小鼠的脚踝上做一个标记，并将右后爪浸入65摄氏度的热水中三秒钟。烧伤后，将鼠标放在干净的家庭笼子中的加热垫上，直到它从麻醉中恢复。将小鼠带到行为测试室，让它们在测试前至少适应30分钟。当老鼠适应环境时，穿上手术服，手套和口罩。接下来，将小鼠放入金属网地板上的方形盒子中，让它们再适应30分钟。然后使用上升刺激法评估后爪的机械阈值，以五到八秒的间隔用一系列Von Frey细丝轻轻戳爪子，以刺激后足。进行五次试验以评估每个同侧后爪的机械阈值。对于步态分析，每天在步态分析系统中适应小鼠，从烧伤前五天开始，持续10至15分钟。在测试当天，戴上手术服、手套和口罩后，将小鼠带到行为测试室，让它们在测试前适应30分钟，然后启动步态分析程序，点击新建实验菜单，指定保存数据的文件夹。指定后，将最大运行时间设置为5秒，将最大允许速度变化设置为50%接下来，在程序的设置选项卡中，选择已注册的相机并将走道长度设置为30厘米。然后在程序菜单的“获取”选项卡上，选择“打开采集”，并根据状态消息，单击快照背景按钮以获取空走道的背景图像。接下来，单击开始采集按钮，并将鼠标放在左/右横穿走道的入口处。录制将在鼠标自由移动后自动开始。成功录制后，从程序菜单的“获取”选项卡中选择“分类运行”，然后选择要分析的运行并单击“自动分类”按钮。自动分类完成后，在每次运行中去除鼻子，生殖器识别和爪子的错误识别到垃圾数据，然后分析数据。在行为测试室中适应30分钟后，将小鼠放入装有15厘米水的透明Plexiglas圆柱体中15分钟。第二天，在相同条件下再次将鼠标放入圆柱体中并测量静止时间，即鼠标停止攀爬或游泳并且头部漂浮在水面上方时。接下来，为了测量正常的运动功能，将动物放在悬挂在设备底部上方16厘米处的滚动圆柱形平台上。在诱导烧伤之前，让小鼠每天在轮转上训练一次至少五天。给药后，每20分钟进行一次轮转试验，持续两小时，截止时间为两分钟。测量鼠标以每分钟15转的恒定速度在旋转杆上运行而不掉落的时间。与对照组相比，烧伤诱导小鼠的爪子退出阈值在第一天后降低，并持续7天。对乙酰氨基酚给药显著降低了烧伤诱导的爪子戒断阈值的降低。此外，曲线下面积分析表明，对乙酰氨基酚给药显著降低了烧伤引起的机械性异常性疼痛。后爪印面积在引诱烧伤后也明显减少，持续7天。与载体治疗相比，对乙酰氨基酚给药显著改善了后爪印面积。此外，烧伤在第一天后减少了同侧后爪的单一姿势，并且这种减少维持了七天。对乙酰氨基酚给药改善后爪单姿势。在强制游泳试验中，与对照组相比，烧伤诱导的小鼠在受伤后7天的不动时间增加。对乙酰氨基酚给药显著减少了烧伤引起的不动时间增加。在rotrorod测试中，烧伤诱导小鼠的运行时间与烧伤诱导后7天的对照组相似。相比之下，甲氧醬酮治疗小鼠的运行时间显着减少约60分钟，表明这种烧伤方法不会引起运动障碍。施加在每只动物身上的烧伤程度必须保持恒定，以获得准确的结果。本研究中提出的各种行为实验分析方法也可以应用于与疼痛，步态障碍和抑郁样行为相关的其他动物模型。所提出的分析方法可以量化烧伤诱导后的各种症状，这将有助于未来烧伤相关研究和治疗方法的发展。

Research

JoVE Journal

Neuroscience

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

在昆虫单感器记录果蝇按蚊</em

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed ...

科研

神经科学

Low-stress Route Learning Using the Lashley III Maze in Mice

低应力路线学习，在老鼠身上使用拉什利三迷宫

Many behavior tests designed to assess learning and memory in rodents, particularly mice, rely on visual cues, food and/or water deprivation, or other ...

T-maze Forced Alternation and Left-right Discrimination Tasks for Assessing Working and Reference Memory in Mice

T-迷宫强迫交替和评估工作，并参考在小鼠记忆的左，右的歧视任务

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in ...

Membrane Potential Dye Imaging of Ventromedial Hypothalamus Neurons From Adult Mice to Study Glucose Sensing

膜下丘脑腹内侧神经元的电位染料成像从成年小鼠为研究葡萄糖传感

Studies of neuronal activity are often performed using neurons from rodents less than 2 months of age due to the technical difficulties associated with ...

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

测定冷灵敏度和适应小鼠一种简单廉价的方法

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice

Optogenetic 在行为小鼠海马θ振荡中的作用

Extensive data on relationships of neural network oscillations to behavior and organization of neuronal discharge across brain regions call for new tools ...

Intracerebroventricular Treatment with Resiniferatoxin and Pain Tests in Mice

小鼠用树脂叶毒素和疼痛测试进行脑血管内治疗

The transient receptor potential vanilloid type 1 (TRPV1), a thermosensitive cation channel, is known to trigger pain in the peripheral nerves. In ...

Intracranial Pharmacotherapy and Pain Assays in Rodents

颅内药物治疗和啮齿动物疼痛检测

Pain is a salient sensory experience with affective and cognitive dimensions. However, central mechanisms for pain remain poorly understood, hindering the ...

Combining Behavior and EEG to Study the Effects of Mindfulness Meditation on Episodic Memory

结合行为和脑电图研究正念冥想对情景记忆的影响

Although there has been recent interest in how mindfulness meditation can affect episodic memory as well as brain structure and function, no study has ...

Dural Stimulation and Periorbital von Frey Testing in Mice As a Preclinical Model of Headache

小鼠硬脑膜刺激和眶周冯弗雷测试作为头痛的临床前模型

The cranial meninges, comprised of the dura mater, arachnoid, and pia mater, are thought to primarily serve structural functions for the nervous system.  ...