Name: comprehensive understanding of inactivity-induced gait alteration in rodents
Uploaded: 2022-07-06T13:10:00
Description: Watch this Scientific Journal Video about Comprehensive Understanding of Inactivity-Induced Gait Alteration in Rodents at JoVE.com

这项研究得到了日本科学促进会（JSPS）KAKENHI（编号18H03129，21K19709，21H03302，15K10441）和日本医学研究与发展机构（AMED）（编号15bk0104037h0002）的部分支持。

本研究已获得京都大学动物实验委员会（Med Kyo 14033）的批准，并符合国立卫生研究院的指导方针（实验动物护理和使用指南，第8版）。7周龄的雄性Wistar大鼠用于本研究。 补充文件1中提供了代表程序顺序的示意图。
1. 让老鼠熟悉跑步机行走
注意：有关该程序的详细信息，请参阅先前发布的报告13 。
<ol><li>将?...

<ol>
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环境的改变导致运动系统的功能方面和肌肉骨骼成分波动26，27。收缩结构或环境中的畸变可能会影响功能能力，即使在解决机械/环境变形后仍然存在19.客观的运动分析有助于定量测量这些功能能力。如上所示，视频分析是获取此类参数的强大方法。
为了跟踪联合地标进行视频分析，使用红外标记和摄像头很普遍?...

缺乏体力活动或废用会导致运动效应器的恶化，例如肌肉萎缩和骨质流失1和全身失调2。此外，最近人们注意到，不活动不仅影响肌肉骨骼组件的结构方面，还影响运动的定性方面。例如，即使在干预结束后1个月，暴露于模拟微重力环境的大鼠的肢体位置也与完整动物的肢体位置不同3，4。然而，关于不活动造成的运动缺陷的报告很少。此外，劣化的综合运动特性尚未完全确定。
目前的协议演示并讨论了运动学评估的应用，通过参考在承受后肢卸载的大鼠中因废弃而引起的步态运动缺陷来可视化运动改变。
已经表明，在模拟微重力环境后行走时四肢的过度伸展在人类5和动物4，6，7，8中都观察到。因此，为了普遍性，我们在本研究中重点关注一般参数：膝关节和踝关节的角度以及跖趾关节与髋关节之间的垂直距离（大致相当于髋关节的高度）在站立阶段（中间）。此外，讨论了视频运动学评估的潜在应用。
一系列运动学分析可能是评估神经控制功能方面的有效措施。然而，尽管运动分析已经从足迹观察或对捕获的视频9，10的简单测量发展到多个相机系统11，12，但通用方法和参数尚未建立。本研究旨在为该关节运动分析提供全面的参数。
在之前的工作13中，我们尝试使用综合视频分析来说明神经病变模型大鼠的步态改变。然而，一般来说，运动分析的潜在结果通常仅限于分析框架中提供的预定变量。出于这个原因，本研究进一步详细说明了如何纳入广泛适用的用户定义的参数。如果实施适当的参数，使用视频分析的运动学评估可能会进一步有用。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Adhesive Tape</td>
			<td>NICHIBAN CO.,LTD.</td>
			<td>SEHA25F</td>
			<td>Adhesive tape to secure thread on tails of rats for hindlimb unloading</td>
		</tr>
		<tr>
			<td>Anesthetic Apparatus for Small Animals</td>
			<td>SHINANO MFG CO.,LTD.</td>
			<td>SN-487-0T</td>
			<td></td>
		</tr>
		<tr>
			<td>Auto clicker</td>
			<td>N.A.</td>
			<td>N.A.</td>
			<td>free software available to download to PC (https://www.google.com/search?client=firefox-b-1-d&#38;q=auto+clicker)</td>
		</tr>
		<tr>
			<td>CCD Camera</td>
			<td>Teledyne FLIR LLC</td>
			<td>GRAS-03K2C-C</td>
			<td>CCD (Charge-Coupled Device) cameras for video capture</td>
		</tr>
		<tr>
			<td>Cotton Thread</td>
			<td>N.A.</td>
			<td>N.A.</td>
			<td>Thread to hang tails of rats from the ceiling of cage</td>
		</tr>
		<tr>
			<td>ISOFLURANE Inhalation Solution</td>
			<td>Pfizer Japan Inc.</td>
			<td>(01)14987114133400</td>
			<td></td>
		</tr>
		<tr>
			<td>Joint marker</td>
			<td>TOKYO MARUI Co., Ltd</td>
			<td>0.12g BB</td>
			<td>6 mm airsoft pellets that were used as semispherical markers with modification</td>
		</tr>
		<tr>
			<td>Kine Analyzer</td>
			<td>KISSEI COMTEC CO.,LTD.</td>
			<td>N.A.</td>
			<td>Software for analysis</td>
		</tr>
		<tr>
			<td>Konishi Aron Alpha</td>
			<td>TOAGOSEI CO.,LTD.</td>
			<td>#31204</td>
			<td>Super glue to attach spherical markers on randmarks of rats</td>
		</tr>
		<tr>
			<td>Motion Recorder</td>
			<td>KISSEI COMTEC CO.,LTD.</td>
			<td>N.A.</td>
			<td>Software for video recording</td>
		</tr>
		<tr>
			<td>Paint Marker</td>
			<td>MITSUBISHI PENCIL CO., LTD</td>
			<td>PX-21.13</td>
			<td>Oil based paint marker to mark toes of animals</td>
		</tr>
		<tr>
			<td>Three-dimensional motion capture apparatus (KinemaTracer for small animals)</td>
			<td>KISSEI COMTEC CO.,LTD.</td>
			<td>N.A.</td>
			<td>3D motion analysis system that consists of four cameras (https://www.kicnet.co.jp/solutions/biosignal/animals/kinematracer-for-animal/ or https://micekc.com/en/)</td>
		</tr>
		<tr>
			<td>Three-dimensional(3D) Calculator</td>
			<td>KISSEI COMTEC CO.,LTD.</td>
			<td>N.A.</td>
			<td>Software fo marker tracking</td>
		</tr>
		<tr>
			<td>Treadmill</td>
			<td>MUROMACHI KIKAI CO.,LTD</td>
			<td>MK-685</td>
			<td>Treadmill equipped with transparent housing, electrical shocker, and speed control unit</td>
		</tr>
		<tr>
			<td>Wistar Rats (male, 7-week old)</td>
			<td>N.A.</td>
			<td>N.A.</td>
			<td>Commercially available at experimental animal sources</td>
		</tr>
	</tbody>
</table>

comprehensive understanding of inactivity-induced gait alteration in rodents

众所周知，废弃会影响神经系统，关节运动会发生变化;然而，哪些结局正确表现出这些特征尚不清楚。本研究描述了一种运动分析方法，该方法利用视频捕获的三维（3D）重建。使用这项技术，在暴露于模拟微重力环境的啮齿动物中，通过尾巴卸载后肢，观察到了废弃引起的行走性能的改变。卸货2周后，大鼠在跑步机上行走，用四个电荷耦合器件（CCD）相机捕捉它们的步态运动。使用图像处理软件重建3D运动曲线并与对照受试者的运动曲线进行比较。重建的结局测量成功地描绘了畸形步态运动的不同方面：膝关节和踝关节的过度伸展以及站立阶段髋关节的较高位置。运动分析很有用，原因有几个。首先，它支持定量行为评估而不是主观观察（例如，在某些任务中通过/失败）。其次，一旦获得基本数据集，就可以提取多个参数以满足特定需求。尽管存在更广泛应用的障碍，但通过确定综合测量和实验程序，可以减轻该方法的缺点，包括劳动强度和成本。

12只动物被随机分配到两组之一：卸载组（UL，n = 6）或对照组（Ctrl，n = 6）。对于UL组，动物的后肢被尾巴卸载2周（UL期），而Ctrl组动物则保持自由。卸载 2 周后，与 Ctrl 组相比，UL 组表现出明显的步态模式。 图1 显示了代表性受试者的归一化关节轨迹。在站立阶段，UL组在膝盖和脚踝的伸展（即脚踝的跖屈）比Ctrl组更突出，称为“脚趾行走”3，<...

Watch this Scientific Journal Video about Comprehensive Understanding of Inactivity-Induced Gait Alteration in Rodents at JoVE.com

Comprehensive Understanding of Inactivity-Induced Gait Alteration in Rodents

本协议描述了三维运动跟踪/评估，以描绘暴露于模拟废弃环境后大鼠的步态运动改变。

全面了解啮齿动物不活动引起的步态改变

It is well known that disuse affects neural systems and that joint motions become altered; however, which outcomes properly exhibit these characteristics ...

我们的协议提供了一个全面的程序来确定关节运动分析的参数。由于缺乏全面的参数定义，关节运动分析通常很复杂。我们展示了普遍适用于相关领域的参数定义的标准原理图。首先，打开运动记录器应用程序。将老鼠放在跑步机皮带上。将皮带速度提高到每秒 20 厘米。当老鼠开始以所需的速度正常行走时，单击 记录 图标开始视频捕获。获得 5 到 10 个连续步骤后，通过单击 记录 图标。打开3DCalculator应用程序，以及要分析的视频文件。通过调整顶部的水平滑块来裁剪视频，以包含足够数量的连续步长。要捕获标记，请将简笔画模型上的标记图例拖动到捕获的视频上的相应标记，然后松开按钮。单击自动跟踪图标。如果系统无法准确跟踪标记，或者跟踪过程因标记丢失而停止，请单击手动图标切换到手动模式。单击简笔图片上缺少的标记图例，以及视频上的相应标记。打开 KineAnalyzer 应用程序并加载文件。转到“查看”，然后单击“编辑标记母版”菜单。它将打开标记主编辑窗口。单击标记“选项卡上所需的标签或地标，然后单击所需的颜色。此过程将每个标记指定为特定地标。然后在链接“选项卡上，通过连续单击两个标记来创建对应于每个肢体的线条。通过从“颜色”列中选择所需的颜色，为创建的线条指定颜色。在角度“选项卡上定义角度。命名角度后，通过单击与每个地标对应的标记，将矢量 A 指定为参考线，将矢量 B 指定为移动线。然后在同一选项卡的“操作”部分中使用值定义角度的方向。接下来，在距离“选项卡上，定义距离参数。在距离设置部分中选择两个相应的标记。每只六只动物被随机分配到卸载组，其中动物的后肢被尾巴卸载两周，或对照组，其中动物被释放。站立阶段膝关节和踝关节的归一化轨迹显示，与对照组相比，卸载组在膝关节和踝关节的伸展程度更高。卸荷后2周，空载组中位膝角、踝角等定量指标显著大于对照组。跖骨-髋关节距离几乎相当于站立中期髋关节的高度，在空载组中也明显更高。这种方法为研究人员提供了如何确定关节运动参数的基础知识。通过这种方式，他们可以建立特定的框架来解决自己的研究问题。

Research

JoVE Journal

Neuroscience

Preparing Undercut Model of Posttraumatic Epileptogenesis in Rodents

准备在鼠害外伤性癫痫的底切模型

Partially isolated cortex ("undercut") is an animal model of posttraumatic epileptogenesis. The surgical procedure involves cutting through the ...

科研

神经科学

Habituation and Prepulse Inhibition of Acoustic Startle in Rodents

在鼠害声不惊人死不休的习惯和前脉冲抑制

The acoustic startle response is a protective response, elicited by a sudden and intense acoustic stimulus. Facial and skeletal muscles are activated ...

Large-scale Recording of Neurons by Movable Silicon Probes in Behaving Rodents

移动硅探针的神经行为鼠害的大型记录

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of ...

A Low Cost Setup for Behavioral Audiometry in Rodents

在啮齿动物的行为测听的低成本设置

In auditory animal research it is crucial to have precise information about basic hearing parameters of the animal subjects that are involved in the ...

Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

运行轮活动在啮齿类动物的昼夜节律的记录和分析

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, ...

Deep Brain Stimulation with Simultaneous fMRI in Rodents

深部脑刺激与同步功能磁共振成像在鼠害

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for ...

Acrylic Resin Molding Based Head Fixation Technique in Rodents

在鼠害丙烯酸树脂模塑基头固定技术

Head fixation is a technique of immobilizing animal's head by attaching a head-post on the skull for rigid clamping. Traditional head fixation requires ...

Transcranial Electrical Brain Stimulation in Alert Rodents

警惕啮齿动物的经颅电脑刺激

Transcranial electrical brain stimulation can modulate cortical excitability and plasticity in humans and rodents. The most common form of stimulation in ...

Automated Gait Analysis in Mice with Chronic Constriction Injury

慢性压迫性损伤小鼠步态的自动分析

The von Frey test is a classical method that has been widely used to examine the sensory function of neuropathic pain animals. However, it has some ...

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 ...