转棒实验用于评估偏侧帕金森综合征大鼠行走运动中的运动功能状态。
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勘误通知
Important: There has been an erratum issued for this article. View Erratum Notice
摘要
摘要
高速视频分析可以提供有关行走运动定性特征的详细信息。当动物在转棒上行走时，其位置基本保持不变，进行重复的踏步运动。因此，该任务能够提供丰富的足部踏步运动细节信息。实验对象为偏侧帕金森大鼠模型，通过向右侧黑质纹状体束注射6-羟基多巴胺（6-OHDA）以耗竭黑质纹状体多巴胺（DA）而制备。本报告展示了此前从前方、侧面和后方视角拍摄的动物行走视频的分析示例（15）。通过评分量表及对踏步行为视频记录的逐帧回放分析发现，偏侧帕金森大鼠在多巴胺耗竭对侧的姿势控制和肢体使用方面存在慢性功能障碍。与同侧肢体相比，对侧肢体在启动和维持推进运动中的参与程度较低。这种由单侧多巴胺耗竭引起的缺陷表明，转棒实验可作为分析踏步运动的有效任务。
方案
转棒行走装置
转棒仪是一个直径为4 cm的旋转圆柱体，固定在距地面35 cm的高度，周围由透明有机玻璃包围。转棒仪的圆柱体表面覆盖有纹理橡胶涂层，以增强抓地力。仪器底部铺有一层泡沫材料，以防止动物跌落时受伤。小型电动机通过橡胶皮带驱动转棒旋转。转棒设定为每5秒旋转一圈。
视频记录与运动学分析
使用高速数码摄像机拍摄动物运动，快门速度为每秒1/1000。在动物于转棒上行走时，分别从前方、侧面和后方进行拍摄。记录过程中采用尼康双臂MII冷光源及一套光纤照明设备，以提供充足的光照条件。录像通过索尼DSR 20 DV CAM录像机逐帧进行分析。视频帧使用图像采集卡和Macintosh计算机进行捕获。
测试步骤与分析
为了评估6-OHDA模型大鼠与对照组大鼠在行走运动上的差异，两名独立观察者在不知晓实验分组的情况下，对术后第15至30天（此时损伤后的恢复过程基本已完成）的视频记录进行了分析。黑质纹状体的6-OHDA损伤诱导于右侧半球，导致左侧前肢和后肢出现持续性功能障碍。研究采用了一种评分量表来评估大鼠的姿势以及前肢的迈步动作（Whishaw等，2003）。迈步动作的评估包含以下五个方面：
- 姿势，通过观察大鼠在滚筒上的位置来....
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讨论
本研究的目的是探讨用于啮齿类动物运动技能检测的转棒实验是否可用于神经系统损伤大鼠姿势、前肢迈步和后肢迈步的定性评估。在分析过程中，对单侧多巴胺（DA）耗竭的大鼠和对照组大鼠分别从前方、侧方和后方进行视频记录。姿势与前肢运动的评分量表显示，手术后大鼠的迈步运动长期受损。对肢体运动的进一步分析表明，尽管多巴胺耗竭大鼠能够使用病灶对侧肢体进行支撑，但在转移体重方面几乎无法有效利用。本研究结果表明，转棒实验除了可提供运动功能障碍的定量指标外，还能揭示其定性方面的功能缺陷。
转棒实验已被广泛用于评估啮齿类动物的运动功能。迄今为止，该实验主要采用终点指标，即动物能够行走的速度或持续时间。我们认为，该实验的某些特性可能使其非常适合对行走动作进行定性分析。首先，由于动物必须保持平衡并进行踏步，该任务涉及较高的技能要求，因此可能揭示出在简单的地面行走任务中不易察觉的功能障碍（Schallert et al., 1992；Olsson et al., 1995；Chang et al., 1999；Muir 和 Whishaw 1999b；Metz et al., 2005）。该任务及其视频分析方法.......
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致谢
由加拿大卫生研究院、加拿大卒中网络以及阿尔伯塔医学研究遗产基金会的资助支持。
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参考文献
- Chang, J. W., Wachtel, S. R., Young, D., Kang, U. J. Biochemical and anatomical characterization of forepaw adjusting steps in rat models of Parkinson's disease: studies on medial forebrain bundle and striatal lesions. Neuroscience. 88, 617-628 (1999).
- Metz, G. A., Tse, A., Ballermann, M., Smith, L. K., Fouad, K.
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重印与许可
勘误
Formal Correction: Erratum: Use of Rotorod as a Method for the Qualitative Analysis of Walking in Rat
Posted by JoVE Editors on 4/01/2012. Citeable Link.
A correction was made to: Use of Rotorod as a Method for the Qualitative Analysis of Walking in Rat. An additional paragraph was added to the discussion, the abstract was modified, and another reference was included.
Additional reference:
15. Whishaw IQ, Li K, Whishaw PA, Gorny B, Metz GA. Distinct forelimb and hind limb stepping impairments in unilateral dopamine-depleted rats: use of the rotorod as a method for the qualitative analysis of skilled walking. J Neurosci Meth 126, 13-23 (2003).
Revised Abstract:
High speed videoanalysis of the details of movement can provide a source of information about qualitative aspects of walking movements. When walking on a rotorod, animals remain in approximately the same place making repetitive movements of stepping. Thus the task provides a rich source of information on the details of foot stepping movements. Subjects were hemi-Parkinson analogue rats, produced by injection of 6-hydroxydopamine (6-OHDA) into the right nigrostriatal bundle to deplete nigrostriatal dopamine (DA). The present report provides a video analysis illustration of animals previously were filmed from frontal, lateral, and posterior views as they walked (15). Rating scales and frame-by-frame replay of the video records of stepping behavior indicated that the hemi-Parkinson rats were chronically impaired in posture and limb use contralateral to the DA-depletion. The contralateral limbs participated less in initiating and sustaining propulsion than the ipsilateral limbs. These deficits secondary to unilateral DA-depletion show that the rotorod provides a use task for the analysis of stepping movements.
Additional Discussion Paragraph:
A more detailed presentation of the present study has been made (Whishaw et al, 2003), but the present study presents the video support describing the stepping movement in the good and affected limbs of unilateral dopamine-depleted rats. For the analysis, rats with unilateral DA depletions and control rats were video recorded from front, lateral and posterior views. A rating scale of posture and forelimb movements indicated that stepping movements were chronically impaired following surgery. Examination of limb movements indicated that whereas the DA-depleted rats could use the limbs contralateral to the lesion for support, they received minimal use for shifting weight. The results of this study indicate that the rotorod task, in addition to providing quantitative measures of motor impairments, can also provide insights into the qualitative impairments.
Original Abstract:
The rotorod test, in which animals walk on a rotating drum, is widely used to assess motor status in laboratory rodents. Performance is measured by the duration that an animal stays up on the drum as a function of drum speed. Here we report that the task provides a rich source of information about qualitative aspects of walking movements. Because movements are performed in a fixed location, they can readily be examined using high-speed video recording methods. The present study was undertaken to examine the potential of the rotorod to reveal qualitative changes in the walking movements of hemi-Parkinson analogue rats, produced by injection of 6-hydroxydopamine (6-OHDA) into the right nigrostriatal bundle to deplete nigrostriatal dopamine (DA). Beginning on the day following surgery and then periodically over the next two months, the rats were filmed from frontal, lateral, and posterior views as they walked on the rotorod. Behavior was analyzed by frame-by-frame replay of the video records. Rating scales of stepping behavior indicated that the hemi-Parkinson rats were chronically impaired in their posture and in the use of the limbs contralateral to the DA-depletion. The contralateral limbs not only displayed postural and movement abnormalities, they participated less in initiating and sustaining propulsion than did the ipsilateral limbs. These findings not only reveal new deficits secondary to unilateral DA-depletion, but also show that the rotorod can provide a robust tool for the qualitative analysis of movement.
