Name: behavioral assessments of spontaneous locomotion in a murine mptp-induced parkinson's disease model
Uploaded: 2019-01-07T14:00:00
Description: Watch this Scientific Journal Video about Behavioral Assessments of Spontaneous Locomotion in a Murine MPTP-induced Parkinson's Disease Model at JoVE.com

我们的工作由天津创新和创业本科培训项目 (批准号 63183004) 资助。该项目是在南开大学药用化学生物学国家重点实验室启动的。提交人声明不存在利益冲突。

这项研究是根据考虑动物实验的国际、国家和机构规则进行的。研究方案得到了南开大学动物伦理委员会的批准。
1. mptp 和 l-dopa 管理
注: 中国科学院动物学研究所提供了为期10周的雌性 balb1 小鼠。在12小时的光/暗周期 (在08岁至20:00 亮起)、21-22°c 的恒温和55% ±5% 的相对湿度下, 每个笼子容纳了6个小鼠。对所有动物给予同一配方的高压灭菌标准小鼠和水</...

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由于大脑黑质中多巴胺能神经元的破坏, mptp 在小鼠7中引起类似帕金森症的症状。l-dopa 是自临床使用以来最受欢迎的 pd 药物, 因为它有助于维持 pd 患者的正常日常活动, 有效抑制运动异常, 包括亚术和僵硬15。用 mptp 治疗的小鼠在圆筒试验和开放场试验等行为测试中表现出损伤, 可用于 mptp 模型的研究。气缸试验和露天试验通常用于测量自发作用。在这里, 我们?...

帕金森病 (pd) 是老年人中最常见的疾病之一, 是一种长期的神经退行性疾病1.患者总是表现出随着时间的推移而恶化的晃动、僵硬、运动缓慢和痴呆的现象.其他症状, 包括感官、睡眠和情绪问题也被观察到2。pd 的原因尚不清楚, 但一般认为它既涉及遗传因素, 也涉及环境因素, 这些因素导致黑质3中多巴胺能神经元的丢失, 以及黑质中莱维体和莱维神经元的发育。大脑4.
在 pd 的研究中, 1-甲基-4-苯基-1、2、3、6-四氢吡啶 (mptp)5在实验模型中被广泛用于再现一些 pd 症状。1984年, langston等人首次发现, 在松鼠猴身上注射 mptp 导致帕金森病.尽管 mptp 啮齿类动物模型没有显示 lewy 体的存在, 这是 pd 的生物标志物, 但 mptp 通过破坏大脑黑质7中的多巴胺能神经元而引起帕金森病样症状.与其他 pd 药物模型 (如 6-羟基多巴胺 (6-ohda)8和 1-甲基-4-苯基吡啶 (mpp +)9诱导的药物模型相比, mptp 注射操作容易, mptp 模型所需时间更短。小鼠接受7天的 mptp 注射 (20-30 mgggg"天), 并在第八天第10日进行行为测试。
公开场试验1 1 首次由美国1 2号的卡尔文 s. 霍尔开发。在各种研究中, 对不同类型的行为进行了测试。在以帕金森病为重点的研究中, 对运动活动和运动速度等行为进行了测试, 以确定动物的移动能力是否受到影响。与其他用于测试 pd 动物建立的方法相比, 开放现场测试很容易进行, 因为所需的设备简单, 原型和数据分析软件 (如matlab、excel) 可以很容易地收集和绘制数据。另外, 变异系数相对较小, 为 13,这意味着露天试验的结果是可靠的。与其他方法相比, 另一个优点是本实验中包含的行为易于区分;老鼠可以是跑的, 也可以是站着的。通常, 当研究人员需要评估研究对象的流动性时, 开放场测试可以用于老鼠。
气缸试验也被称为不对称使用前肢的试验。在首次设计该测试时, 它被用来测试老鼠前肢14的不对称使用。在这里, 我们用这个测试来分析动物的能力, 伸展和使用它的两个前肢探索新的环境。当黑质和纹状体在大脑中被 mptp 破坏时, 动物往往会弓起背部, 更不可能伸展和探索未知的环境。此测试易于执行, 并能给出初步结果。然而, 这种测试具有很高的内部变异性, 所以它通常与其他行为实验一起使用。
以 l-dopa (也称为左旋多巴或 l-3, 000-二羟基苯基丙氨酸) 为治疗帕金森病的常用方法, 因为 pd 的一个原因是多巴胺的减少。l-dopa 是多巴胺的前体。但与多巴胺不同的是, 它可以跨越血脑屏障, 这意味着它将更有效地增加大脑区域的多巴胺浓度。通过血脑屏障后, l-dopa 被 l-氨基酸脱羧酶15转化为多巴胺。
本文介绍了利用圆筒试验14和改进的开放场试验对 mptp 诱导 pd 模型小鼠运动功能的测量和分析。我们以 l-dopa 为例, 说明了如何将该模型应用于 pd 药物的研究。我们的研究结果表明, mptp 会导致电机赤字, 可以通过 l-dopa 来缓解。

<table>
	<tbody>
		<tr>
			<td>70% Ethanol</td>
			<td>Ruiboxing Company</td>
			<td>RBX-64175</td>
			<td></td>
		</tr>
		<tr>
			<td>Camera</td>
			<td>BASLER</td>
			<td>acA645-100gm</td>
			<td></td>
		</tr>
		<tr>
			<td>Cylinder test</td>
			<td>Made in-house at Nankai University</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Excel</td>
			<td>Microsoft</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Levodopa</td>
			<td>Sigma-Aldrich</td>
			<td>72816</td>
			<td></td>
		</tr>
		<tr>
			<td>Matalb 2017a</td>
			<td>Mathworks</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Mice</td>
			<td>Institute of Zoology, Chinese Academy of Sciences</td>
			<td>Balb/c</td>
			<td>Adult female mice(10 weeks)</td>
		</tr>
		<tr>
			<td>MPTP</td>
			<td>Yuanye Biological Technology Company Ltd., Shanghai</td>
			<td>S31504-500mg</td>
			<td></td>
		</tr>
		<tr>
			<td>Open field test</td>
			<td>Made in-house at Nankai University</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe</td>
			<td>Solelybio</td>
			<td>S-xsgwz-w</td>
			<td>Irrigation</td>
		</tr>
		<tr>
			<td>Syringe</td>
			<td>Jiangxi Fenglin Medical Application Co.</td>
			<td>hc3824</td>
			<td>Intraperitoneal injection</td>
		</tr>
	</tbody>
</table>

behavioral assessments of spontaneous locomotion in a murine mptp-induced parkinson's disease model

帕金森病 (pd) 是一种常见的神经退行性疾病, 引起晃动、僵硬、运动缓慢和痴呆的现象。1-甲基-4-苯基-1, 2, 3, 6-四氢吡啶 (mptp) 可以通过破坏大脑黑质中的多巴胺能神经元导致一些帕金森病样症状。因此, 它被用于在各种动物研究中建立 pd 模型。在这里, 小鼠接受7天的 mptp 注射 (20 mgsg"), 并在第八天进行行为测试。该模型在 pd 研究中得到了有效的适应。这里的行为测试包括气缸测试和开放场测试。气缸实验是用来检测动物的能力, 以提高他们的前爪, 当投入到不同的环境。当 pd 模型小鼠显示拱形--小鼠拱起它的背部--时, 爪子的增加次数就会减少。此测试易于执行。开放场测试用于检测老鼠在跑步、行走和保持不动上的时间。我们使用软件分析动物在野外的运动, 并获得数据。最后, 我们使用 l-dopa, 最常用的 pd 药物之一, 作为一个例子, 说明如何应用该模型的 pd 药物的研究。我们的研究结果表明, mptp 神经毒性会诱发运动缺陷, 而 l-dopa 可以缓解这种情况。

在圆筒试验中, 在第1天至第7天用 mptp 处理的小鼠 (第2组) 和第8天用盐水处理的小鼠 (第1组) 中观察到, 与受过盐水处理的小鼠 (第1组) 相比, 在使用 mp 处理的小鼠 (第3组) 中观察到了更多的环 (第3组)从第1天到第7天和第8天 l-dopa 与第1天至第7天用 mptp 处理的小鼠 (第2组) 和第8天用盐水治疗的小鼠 (第2组) 进行比较 (图 1)。
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Watch this Scientific Journal Video about Behavioral Assessments of Spontaneous Locomotion in a Murine MPTP-induced Parkinson's Disease Model at JoVE.com

Behavioral Assessments of Spontaneous Locomotion in a Murine MPTP-induced Parkinson's Disease Model

小鼠 mptp 诱导的帕金森病模型中自发性运动的行为评估

我们描述了使用 mptp 建立帕金森病小鼠模型, 以及使用圆筒和开放场测试测量运动功能的行为评估。然后, 我们使用 l-dopa 作为一个例子, 说明如何将这个模型应用于 pd 药物的研究。

Parkinson&#8217;s disease (PD) is a common neurodegenerative disorder disease, causing the phenomenon of shaking, rigidity, slowness of movement and ...

在这段视频中，我们介绍了如何使用气缸和开放场测试来缩小帕金森病MPTP模型中的电机功能。主要优点是，在行为测试之前，不需要训练老鼠。此外，该方法效率高，设备成本低。对行为测试的新鲜人可能低估了环境干扰的影响。保持安静和避免衬里的变化会有所帮助。在这里，行为测试进行24小时后，最后注射7天的MPTP方案或盐水注射和40分钟后，胃内给发L-DOPA的一部分动物。如果不在动物居住的同一房间进行行为测试，请让小鼠在测试前 30 到 60 分钟适应新房间。首先将透明玻璃缸放在桌子的中心。用黑色纸板将气缸的三面环绕，放置在离气缸四到八厘米远的地方，以减少环境照明的影响。让气缸的一侧朝向摄像机进行录像。连接距离气缸至少 100 万像素分辨率 40 至 60 厘米的摄像机，以确保整个圆柱体包含在视野中。一次将一只鼠标放入圆柱体中，然后立即开始拍摄。在这个过程中，尽量避免噪音或光线变化，以避免影响鼠标的行为。当鼠标在后肢上抬起后肢时， 爪子升降机会发生， 同时将前肢抬高到肩部以上， 然后降落。三分钟后停止拍摄，把鼠标放回家里的笼子里。用水清洁圆柱体，然后在内壁上喷洒70%的种族，以消毒并去除老鼠的气味。在测试下一个鼠标之前，请擦干气缸。测试完成后，以正常速度的 0.5 倍的速度播放视频，并计算每只鼠标墙上的爪子升降机数量。如果鼠标连续几次在肩部以上提升前肢而不着陆，则只计算一次。在与气缸测试相同的会话期间执行开放场测试。准备一个透明的开放式现场反应箱与木板，覆盖对比黑色或白色布，这取决于鼠标的颜色。将摄像机固定到一米高的场地上。调整摄像机以确保打开的场反应框位于视频的中心。将一只测试鼠标放入框中，让鼠标熟悉环境约一分钟。使用连接到计算机的摄像机录制五分钟的视频。最后，清理露天反应箱中的任何粪便，在箱体上喷洒70%乙醇，然后擦拭。在气缸测试中，与盐水处理的小鼠相比，使用MPTP治疗的小鼠的后壁数量减少。与接受盐水的MPTP治疗小鼠相比，在测试当天给予L-多巴胺的MTPT治疗小鼠中观察到后方增加。此图像显示了未使用 MPTP 治疗的对照小鼠中的代表性运动轨迹和静态时间、行走时间和运行时间分布情况。使用MTPT治疗的小鼠运动速度较低，运动距离更短，静态时间更长，运行时间更短，然后盐水治疗小鼠，表明MTPT引起的运动缺陷。MPTP治疗的小鼠接受L-多巴胺显示，与仅接受MPTP治疗的小鼠，运动速度更高，运动距离更长，静态时间更短，运行时间更长，这表明L-多巴胺缓解了MPTP引起的运动缺陷。当改变被测试的小鼠时，完全清除区域并喷洒乙醇，以消除前小鼠气味的影响。按照此程序，还可以执行其他措施，如光束行走测试、杆子测试。通过这些方法可以获得与平衡能力和运动能力相关的数据。MPTP对小鼠和人类都是神经毒性，因此必须采取严格的预防措施以避免接触。应佩戴额外的 PPE，如面罩或呼吸器。

Research

JoVE Journal

Behavior

Assessment of Sensorimotor Function in Mouse Models of Parkinson's Disease

帕金森氏症小鼠模型的感觉功能评估

Sensitive and  reliable behavioral outcome measures are essential to the evaluation of  potential therapeutic treatments in preclinical trials for many  ...

科研

行为学

Behavioral Phenotyping of Murine Disease Models with the Integrated Behavioral Station (INBEST)

Behavioral Phenotyping of Murine Disease Models with the Integrated Behavioral Station INBEST

小鼠疾病模型的集成行为车站行为表型（INBEST）

Due to rapid advances in genetic engineering, small rodents have become the preferred subjects in many disciplines of biomedical research. In studies of ...

Compensatory Limb Use and Behavioral Assessment of Motor Skill Learning Following Sensorimotor Cortex Injury in a Mouse Model of Ischemic Stroke

补偿肢使用和运动技能学习行为评估以下感觉运动皮层损伤缺血性脑卒中的小鼠模型

Mouse models have become increasingly popular in the field of behavioral neuroscience, and specifically in studies of experimental stroke. As models ...

A Procedure to Observe Context-induced Renewal of Pavlovian-conditioned Alcohol-seeking Behavior in Rats

一个过程来观察大鼠巴甫洛夫空调酒精寻求行为的背景下引发的重建

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate ...

Community-based Adapted Tango Dancing for Individuals with Parkinson's Disease and Older Adults

Adapted tango dancing improves mobility and balance in older adults and additional populations with balance impairments. It is composed of very simple ...

Asymmetric Walkway: A Novel Behavioral Assay for Studying Asymmetric Locomotion

非对称走道：一种新的行为检测方法的研究不对称步态

Behavioral assays are commonly used for the assessment of sensorimotor impairment in the central nervous system (CNS). The most sophisticated methods for ...

Battery of Behavioral Tests Assessing General Locomotion, Muscular Strength, and Coordination in Mice

用于评估一般运动、肌肉力量和小鼠协调性的行为测试电池

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects and outcomes that a particular disease or treatment has on the ...

Detecting Behavioral Deficits in Rats After Traumatic Brain Injury

颅脑损伤后大鼠行为缺陷的检测

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, ...

Automated, Long-term Behavioral Assay for Cognitive Functions in Multiple Genetic Models of Alzheimer's Disease, Using IntelliCage

基于 IntelliCage 的阿尔茨海默病多基因模型认知功能的自动、长期行为检测

Multiple factors&#8212;such as aging and genes&#8212;are frequently associated with cognitive decline. Genetically modified mouse models of cognitive ...