Name: applying the ratwalker system for gait analysis in a genetic rat model of parkinson's disease
Uploaded: 2021-01-18T14:00:00
Description: Watch this Scientific Journal Video about Applying the RatWalker System for Gait Analysis in a Genetic Rat Model of Parkinson's Disease at JoVE.com

KS和HF感谢Michael J Fox帕金森研究基金会对他们在帕金森病方面的工作的支持。

所有动物研究均由内布拉斯加大学医学中心机构动物护理和使用委员会（IACUC）批准。
1. 步态仪
注意：以MouseWalker14为模型，RatWalker设计的尺寸与大鼠和小鼠之间的步长差异成正比。它由侧面照明背光、走道外壳、光波导走道、镜子和摄像头组成（图 S1）。在人行道和背光波导的每一侧都使用了交错定位的 LED 灯条，以容纳额?...

<ol>
	<li>Behari, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parkinson's+disease.">Parkinson's disease.</a> Annals of Indian Academy of Neurology. 14, 2-6 (2011).</li><li>Kalia, L. V., Lang, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parkinson's+disease.">Parkinson's disease.</a> Lancet. 386 (9996), 896-912 (2015).</li><li>Fahn, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Description+of+Parkinson's+disease+as+a+clinical+syndrome.">Description of Parkinson's disease as a clinical syndrome.</a> Annals of the New York Academy of Sciences. 991, 1-14 (2003).</li><li>Frenkel-Toledo, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+gait+speed+on+gait+rhythmicity+in+Parkinson's+disease:+variability+of+stride+time+and+swing+time+respond+differently.">Effect of gait speed on gait rhythmicity in Parkinson's disease: variability of stride time and swing time respond differently.</a> Journal of NeuroEngineering and Rehabilitation. 2, 23 (2005).</li><li>Shulman, J. M., De Jager, P. L., Feany, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parkinson's+disease:+genetics+and+pathogenesis.">Parkinson's disease: genetics and pathogenesis.</a> Annual Review of Pathology. 6, 193-222 (2011).</li><li>Klemann, C., Martens, G. J. M., Poelmans, G., Visser, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Validity+of+the+MPTP-Treated+Mouse+as+a+Model+for+Parkinson's+Disease.">Validity of the MPTP-Treated Mouse as a Model for Parkinson's Disease.</a> Molecular Neurobiology. 53 (3), 1625-1636 (2016).</li><li>Ungerstedt, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Striatal+dopamine+release+after+amphetamine+or+nerve+degeneration+revealed+by+rotational+behaviour.">Striatal dopamine release after amphetamine or nerve degeneration revealed by rotational behaviour.</a> Acta Physiologica Scandinavian Supplementum. 367, 49-68 (1971).</li><li>Beal, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+models+of+Parkinson's+disease.">Experimental models of Parkinson's disease.</a> Nature Reviews Neuroscience. 2 (5), 325-334 (2001).</li><li>Betarbet, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+systemic+pesticide+exposure+reproduces+features+of+Parkinson's+disease.">Chronic systemic pesticide exposure reproduces features of Parkinson's disease.</a> Nature Neuroscience. 3 (12), 1301-1306 (2000).</li><li>Cannon, J. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+highly+reproducible+rotenone+model+of+Parkinson's+disease.">A highly reproducible rotenone model of Parkinson's disease.</a> Neurobiology of Disease. 34 (2), 279-290 (2009).</li><li>Quary, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Major+strain+differences+in+response+to+chronic+systemic+administration+of+the+mitochondrial+toxin+3-nitropropionic+acid+in+rats:+implications+for+neuroprotection+studies.">Major strain differences in response to chronic systemic administration of the mitochondrial toxin 3-nitropropionic acid in rats: implications for neuroprotection studies.</a> Neuroscience. 97 (3), 521-530 (2000).</li><li>Cicchetti, F., Drouin-Ouellet, J., Gross, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Environmental+toxins+and+Parkinson's+disease:+what+have+we+learned+from+pesticide-induced+animal+models.">Environmental toxins and Parkinson's disease: what have we learned from pesticide-induced animal models.</a> Trends in Pharmacological Sciences. 30 (9), 475-483 (2009).</li><li>Greenamyre, J. T., Cannon, J. R., Drolet, R., Mastroberardino, P. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lessons+from+the+rotenone+model+of+Parkinson's+disease.">Lessons from the rotenone model of Parkinson's disease.</a> Trends in Pharmacological Sciences. 31 (4), 142-143 (2010).</li><li>Mendes, C. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+gait+parameters+in+freely+walking+rodents.">Quantification of gait parameters in freely walking rodents.</a> BMC Biology. 13, 50 (2015).</li><li>Hamers, F. P., Lankhorst, A. J., van Laar, T. J., Veldhuis, W. B., Gispen, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+quantitative+gait+analysis+during+overground+locomotion+in+the+rat:+its+application+to+spinal+cord+contusion+and+transection+injuries.">Automated quantitative gait analysis during overground locomotion in the rat: its application to spinal cord contusion and transection injuries.</a> Journal of Neurotrauma. 18 (2), 187-201 (2001).</li><li>Hamers, F. P., Koopmans, G. C., Joosten, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CatWalk-assisted+gait+analysis+in+the+assessment+of+spinal+cord+injury.">CatWalk-assisted gait analysis in the assessment of spinal cord injury.</a> Journal of Neurotrauma. 23 (3-4), 537-548 (2006).</li><li>Pickrell, A. M., Youle, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+roles+of+PINK1,+parkin,+and+mitochondrial+fidelity+in+Parkinson's+disease.">The roles of PINK1, parkin, and mitochondrial fidelity in Parkinson's disease.</a> Neuron. 85 (2), 257-273 (2015).</li><li>Dawson, T. M., Ko, H. S., Dawson, V. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+animal+models+of+Parkinson's+disease.">Genetic animal models of Parkinson's disease.</a> Neuron. 66 (5), 646-661 (2010).</li><li>Gispert, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parkinson+phenotype+in+aged+PINK1-deficient+mice+is+accompanied+by+progressive+mitochondrial+dysfunction+in+absence+of+neurodegeneration.">Parkinson phenotype in aged PINK1-deficient mice is accompanied by progressive mitochondrial dysfunction in absence of neurodegeneration.</a> PLoS One. 4 (6), 5777 (2009).</li><li>Goldberg, M. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parkin-deficient+mice+exhibit+nigrostriatal+deficits+but+not+loss+of+dopaminergic+neurons.">Parkin-deficient mice exhibit nigrostriatal deficits but not loss of dopaminergic neurons.</a> Journal of Biological Chemistry. 278 (44), 43628-43635 (2003).</li><li>Kitada, T., Tong, Y., Gautier, C. A., Shen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Absence+of+nigral+degeneration+in+aged+parkin/DJ-1/PINK1+triple+knockout+mice.">Absence of nigral degeneration in aged parkin/DJ-1/PINK1 triple knockout mice.</a> Journal of Neurochemistry. 111 (3), 696-702 (2009).</li><li>Dave, K. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phenotypic+characterization+of+recessive+gene+knockout+rat+models+of+Parkinson's+disease.">Phenotypic characterization of recessive gene knockout rat models of Parkinson's disease.</a> Neurobiology of Disease. 70, 190-203 (2014).</li><li>Schindelin, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fiji:+an+open-source+platform+for+biological-image+analysis.">Fiji: an open-source platform for biological-image analysis.</a> Nature Methods. 9 (7), 676-682 (2012).</li><li>Hruska, R. E., Kennedy, S., Silbergeld, E. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+aspects+of+normal+locomotion+in+rats.">Quantitative aspects of normal locomotion in rats.</a> Life Sciences. 25 (2), 171-179 (1979).</li><li>de Medinaceli, L., Freed, W. J., Wyatt, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+index+of+the+functional+condition+of+rat+sciatic+nerve+based+on+measurements+made+from+walking+tracks.">An index of the functional condition of rat sciatic nerve based on measurements made from walking tracks.</a> Experimental Neurology. 77 (3), 634-643 (1982).</li><li>Kunkel-Bagden, E., Dai, H. N., Bregman, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+to+assess+the+development+and+recovery+of+locomotor+function+after+spinal+cord+injury+in+rats.">Methods to assess the development and recovery of locomotor function after spinal cord injury in rats.</a> Experimental Neurology. 119 (2), 153-164 (1993).</li><li>Jacobs, B. Y., Kloefkorn, H. E., Allen, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gait+Analysis+Methods+for+Rodent+Models+of+Osteoarthritis.">Gait Analysis Methods for Rodent Models of Osteoarthritis.</a> Current Pain and Headache Reports. 18 (10), 456 (2014).</li><li>Boix, J., von Hieber, D., Connor, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gait+Analysis+for+Early+Detection+of+Motor+Symptoms+in+the+6-OHDA+Rat+Model+of+Parkinson's+Disease.">Gait Analysis for Early Detection of Motor Symptoms in the 6-OHDA Rat Model of Parkinson's Disease.</a> Frontiers in Behavioral Neuroscience. 12, 39 (2018).</li><li>Zhou, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gait+analysis+in+three+different+6-hydroxydopamine+rat+models+of+Parkinson's+disease.">Gait analysis in three different 6-hydroxydopamine rat models of Parkinson's disease.</a> Neuroscience Letters. 584, 184-189 (2015).</li><li>Vandeputte, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+quantitative+gait+analysis+in+animal+models+of+movement+disorders.">Automated quantitative gait analysis in animal models of movement disorders.</a> BMC Neuroscience. 11, 92 (2010).</li><li>Chuang, C. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+evaluation+of+motor+function+before+and+after+engraftment+of+dopaminergic+neurons+in+a+rat+model+of+Parkinson's+disease.">Quantitative evaluation of motor function before and after engraftment of dopaminergic neurons in a rat model of Parkinson's disease.</a> Journal of Biomedical Science. 17, 9 (2010).</li><li>Baldwin, H. A., Koivula, P. P., Necarsulmer, J. C., Whitaker, K. W., Harvey, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Step+Sequence+Is+a+Critical+Gait+Parameter+of+Unilateral+6-OHDA+Parkinson's+Rat+Models.">Step Sequence Is a Critical Gait Parameter of Unilateral 6-OHDA Parkinson's Rat Models.</a> Cell Transplantation. 26 (4), 659-667 (2017).</li></ol>

步态障碍，包括手臂摆动减少、行走速度减慢和步数缩短，是帕金森病的一个决定性特征，发生在病程早期1，5。多年来，已经开发了几种方法来观察和记录脚步，以便在PD的啮齿动物模型中进行步态分析，手动技术用于量化脚步位置，从而产生了更灵敏且能够捕获动态参数的自动化方法。一些静态方法涉及“墨迹”啮齿动物的爪子，并带有指示器，以?...

PD是最常见的年龄相关性神经退行性运动障碍，是由黑质致密体中DA神经元的缺失引起的。这种黑质DA神经元的缺失和进入纹状体的DA输入导致PD1，2患者观察到的运动功能障碍。帕金森病患者的运动特征（统称为帕金森综合征）包括强直、静止性震颤、运动迟缓、姿势不稳和显微照片3。此外，步态障碍在PD患者中很常见，出现在疾病早期1，4，5。虽然建议采取某些生活方式来帮助减缓PD的进展，例如健康饮食和定期运动，但目前尚无治愈PD的方法，只有药物来控制症状。这为进一步研究留下了空间，以期改进治疗方法。因此，PD动物模型中步态模式的表征是表征模型相关性以及旨在控制PD的治疗如何预防或改善运动障碍的关键工具。
有各种PD动物模型已被用于测试治疗，但是每种模型都有其局限性。例如，用神经毒素1-甲基-4-苯基-1，2，3，6-四氢吡啶（MPTP）处理的动物模型已经产生了大量关于对黑质DA神经元缺失和随后的纹状体适应很重要的过程的信息，并指出线粒体在PD发病机制中的作用;然而，MPTP模型的致病背景具有毒性，而不是像人类PD6那样的神经退行性过程。其他化学诱导模型包括6-羟基多巴胺（6-OHDA）和鱼藤酮。6-OHDA是第一种通过药物在DA神经元中的选择性积累来诱导PD的药物，最终杀死神经元并导致PD样症状。该模型首先用于通过检查对苯丙胺和阿扑吗啡的反应来跟踪DA耗竭7。这种PD诱导方法已被证明可用于筛选影响DA及其受体的药物8。虽然6-OHDA模型是跟踪可量化运动缺陷的一个很好的模型，但该模型并未显示神经元的逐渐丧失和路易体的形成如何影响动物。另一种诱导方法鱼藤酮已被证明具有黑质纹状体神经元的进行性变性，酪氨酸羟化酶和DA转运蛋白的丢失，从而允许更好的模型来跟踪神经元随时间的损失9。鱼藤酮处理的大鼠表现出运动迟缓，姿势不稳定和步态不稳10。然而，已经发现这种方法在不同品系的大鼠之间差异很大，这引发了对鱼藤酮是否是可靠的PD模型的质疑11，12，13。虽然步态分析已被证明受到大鼠PD诱导的影响，但迄今为止，遗传诱导的PD大鼠模型尚未通过自由走在跑道上轻松用于步态分析。
分析自由行走的啮齿动物运动障碍的一种方法是运动学步态分析，可以通过利用FTIR成像进行。这种既定方法使用基于FTIR的光学触摸传感器，该传感器记录和跟踪啮齿动物在跑道14，15，16上移动时的脚印。与其他方法相比，FTIR不依赖于动物身体上任何可能干扰爪印的标记。视频数据的生成产生所有四个肢体的数字爪印，可以组合在一起，为各种啮齿动物模型创建动态且可重复的行走模式。基于成像的步态分析的原理是，当啮齿动物走下跑道时，取下每只爪子并测量随时间推移的接触面积。每种姿势都由爪面积的增加（在制动阶段）和爪面积的减少（在推进阶段）表示。这是通过摆动阶段进行的，即没有检测到爪子信号时。在对视频进行评估后，将生成几个参数，可用于比较野生型（WT）与PD模型。参数的一些示例包括步长（爪子一步覆盖的距离）、摆动持续时间（爪子不与跑道接触的时间持续时间）、摆动速度（步长与摆动持续时间的函数）和步型（对角线步、横向步长或腰带步）。
为了证明FTIR在揭示大鼠早期步态模式变化方面的效用，我们使用了PD的遗传大鼠模型。虽然大多数PD病例是特发性的;PD遗传形式的鉴定发现了基因突变和变异，例如Pink1和Parkin的功能丧失突变，这两种蛋白质参与线粒体质量控制17，可以用来创建动物模型18。不幸的是，小鼠在失去这些蛋白质（单一和组合）时对神经变性具有抗性19，20，21。在大鼠中，Pink1而不是Parkin缺乏导致黑质DA神经元丢失和运动障碍22，但没有完全外显。因此，我们生成了一个组合的Pink1 / Parkin双敲除（DKO）大鼠模型，该模型显示了雄性Pink1 KO大鼠22中报告的明显视觉上明显的后肢拖曳表型，但现在的比率更高：100%对4-6个月之间的雄性30-50%。
虽然这种方法适用于分析小鼠14的运动缺陷，但FTIR成像步态系统规格以适应大鼠的大小和体重以前是非商业性的。在这里，我们解释了如何构建RatWalker，这是一种以MouseWalker14为模型的改进的FTIR步态成像系统，除了适应老鼠的大小和体重。该系统利用光学效应FTIR提供了一种可视化并随后记录动物足迹以进行分析的方法。动物的脚与光波导（平台）接触会导致光路中断，从而产生可见的散射效应，这是使用家用级高速摄像和使用开源软件进行处理的。这项研究证明了FTIR成像在研究PD遗传大鼠模型中步态变化方面的能力。例如，虽然最早在雄性DKO大鼠4个月时观察到明显的视觉上明显的运动变化（即后肢拖拽），但使用FTIR，我们能够在2个月大时发现雄性DKO大鼠的门异常。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Aluminum</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1.5&#8221; Aluminum Angle (1/8&#8221; - 6063)</td>
			<td></td>
			<td></td>
			<td>Dimensions: 8&#39; 
			Qty: 8</td>
		</tr>
		<tr>
			<td>1&#8221; Aluminum Square Tube (1/16&#8221; - 6063)</td>
			<td></td>
			<td></td>
			<td>Dimensions: 8&#39; 
			Qty: 4</td>
		</tr>
		<tr>
			<td>32 Gauge Aluminum Sheet</td>
			<td></td>
			<td></td>
			<td>Dimensions: 10&#39; 
			Qty: 1</td>
		</tr>
		<tr>
			<td>1&#8221; Aluminum Tube (1/8&#8221; - 6063)</td>
			<td></td>
			<td></td>
			<td>Dimensions: 8&#39; 
			Qty: 1</td>
		</tr>
		<tr>
			<td>Acrylic</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>7/32&#8221; Clear Acrylic Sheet</td>
			<td></td>
			<td></td>
			<td>Dimensions: 4&#39;x8&#39; 
			Qty: 2</td>
		</tr>
		<tr>
			<td>1/8&#8221; White Acrylic Sheet 55% (2447)</td>
			<td></td>
			<td></td>
			<td>Dimensions: 4&#39;x8&#39; 
			Qty: 1</td>
		</tr>
		<tr>
			<td>Mirror</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>7/32&#8221; Glass Mirror</td>
			<td></td>
			<td></td>
			<td>Dimensions: 60&#34;x12&#34; 
			Qty: 1</td>
		</tr>
		<tr>
			<td>LED</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>5050 LED Tape Light (Green)</td>
			<td></td>
			<td></td>
			<td>Dimensions: 16.4&#39; 
			Qty: 1</td>
		</tr>
		<tr>
			<td>5050 LED Tape Light (Red)</td>
			<td></td>
			<td></td>
			<td>Dimensions: 16.4&#39; 
			Qty: 1</td>
		</tr>
		<tr>
			<td>Camera</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>GoPro Hero 6 Black</td>
			<td></td>
			<td></td>
			<td>Qty: 1</td>
		</tr>
		<tr>
			<td>Tripod</td>
			<td></td>
			<td></td>
			<td>Dimensions: 57&#34; 
			Qty: 1</td>
		</tr>
	</tbody>
</table>

applying the ratwalker system for gait analysis in a genetic rat model of parkinson's disease

帕金森病 （PD） 是一种进行性神经退行性疾病，由黑质致密部多巴胺能 （DA） 神经元丢失引起。步态异常，包括手臂摆动减少、行走速度减慢和步数缩短，在帕金森病患者中很常见，并且出现在病程早期。因此，PD动物模型中运动模式的定量对于病程和治疗期间的表型表征非常重要。大多数PD病例是特发性的;然而，PD遗传形式的鉴定揭示了基因突变和变异，例如Pink1和Parkin的功能丧失突变，这两种蛋白质参与线粒体质量控制，可以用来创建动物模型。虽然小鼠在失去Pink1和Parkin（单次和组合缺失）时对神经变性具有抵抗力，但在大鼠中，Pink1而不是Parkin缺乏导致黑质DA神经元丧失和运动障碍。在这里，我们报告了FTIR成像的效用，以揭示自由行走的年轻（2个月大）雄性大鼠的步态变化，随着这些大鼠年龄的增长（在4-6个月时观察到），其特征是后肢拖曳，如先前在Pink1敲除（KO）大鼠中报道的那样。

鼠群维护 Pink1和Parkin单KO大鼠的产生和表征已在前面描述过22。Pink1和Parkin单只KO大鼠是从SAGE实验室获得的（现在可以从Envigo获得）。DKO大鼠是通过将Pink1-/-大鼠与Parkin-/-大鼠杂交以获得Pink1 + / / / Parkin+/-大鼠而产生的，这些大鼠杂交以获得Pink1-/-/Parkin-/-大鼠（可从Envigo获得）。为了确认Park6?...

Watch this Scientific Journal Video about Applying the RatWalker System for Gait Analysis in a Genetic Rat Model of Parkinson's Disease at JoVE.com

Applying the RatWalker System for Gait Analysis in a Genetic Rat Model of Parkinson's Disease

在这里，我们描述了RatWalker系统，该系统是通过重新设计MouseWalker设备来构建的，以适应增加的老鼠的尺寸和重量。该系统使用受挫全内反射 （FTIR）、高速视频捕获和开放式访问分析软件来跟踪和量化步态参数。

将RatWalker系统应用于帕金森病遗传大鼠模型中的步态分析

Parkinson's disease (PD) is a progressive neurodegenerative disorder caused by the loss of dopaminergic (DA) neurons in the substantia nigra pars ...

这种方法可以重复地量化步态参数的差异。步态参数可以在自由行走的啮齿动物中进行评估，并且该技术不依赖于侵入性步骤，例如用墨水浸渍脚。演示该程序将是Ben Lamberty。在每次跑步开始之前，首先在人行道的尽头放置一个家庭笼子，作为老鼠穿越人行道的积极提示。关掉房间的灯。打开相机电源并开始记录，然后在将老鼠放在平台上几秒钟。安装围栏后，将老鼠放在人行道的尽头，与家笼相对，并让它以非强迫的方式穿过人行道。一旦动物到达人行道的终点，就停止记录。在跑步之间以及动物排尿或排便后，使用 70% 乙醇和非研磨性毛巾清洁人行道。然后，在引入另一只动物之前让乙醇蒸发。在每个观察期间，大鼠在人行道上总共跑七次，将得分的前三次作为通过进行分析。如果动物向主笼的方向连续四步或更多步，而没有因梳理、暂停或错误的动作而中断，则得分为及格。在两个月龄时对雄性野生型和Parkin DKO大鼠进行步态分析，以确定使用运动学步态分析是否可以在出现粗大运动问题之前发现细微的运动障碍。尽管DKO大鼠的体重增加，但施加在行走表面上的脚压没有改变。在评估几个步态参数作为步行速度的函数时，发现野生型和DKO大鼠的步行速度和步长相似。野生型和DKO大鼠在较慢的步行速度下的站立阶段和摆动持续时间上的变化变得明显。腿处于站立阶段的步进周期部分是占空比。随着占空比的降低，摆动阶段花费的时间多于站立阶段。此外，虽然野生型动物的摆动速度随着速度的增加而增加，但在DKO大鼠中相关性减弱。FTIR步态分析还用于创建自由行走的大鼠每条腿相对于身体的站立阶段曲线图。通过比较爪子位置，观察到AEP和PEP的显着变化。与野生型相比，DKO大鼠的几个附加参数发生了显着变化。DKO大鼠左右后肢摆动速度均增加，左右后肢摆动时间均缩短。设备的一致设置，包括相机相对于RatWalker的位置，对于成功实施该协议非常重要。执行此协议后，动物可用于任何其他必要的行为或实验终点。例如，步态变化可能与组织学改变相关。该技术可用于评估其他非PD啮齿动物模型中的步态，例如焦虑和抑郁模型。

Research

JoVE Journal

Neuroscience

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

可视化和遗传操作在小鼠大脑皮层和使用海马树突和棘<em&gt;在子宫内</em&gt;电穿孔

In utero electroporation (IUE) has become a powerful technique to study the development of different regions of the embryonic nervous system 1-5. To date ...

科研

神经科学

Immunohistochemical Analysis in the Rat Central Nervous System and Peripheral Lymph Node Tissue Sections

免疫组化分析大鼠中枢神经系统和周围淋巴结组织切片

Immunohistochemistry (IHC) provides highly specific, reliable and attractive protein visualization. Correct performance and interpretation of an IHC-based ...

Methods to Characterize Spontaneous and Startle-induced Locomotion in a Rotenone-induced Parkinson's Disease Model of Drosophila

Methods to Characterize Spontaneous and Startle-induced Locomotion in a Rotenone-induced Parkinson's Disease Model of Drosophila

方法来描述自发和惊吓引起的步态中的鱼藤酮诱导的帕金森病模型<em&gt;果蝇</em

Parkinson&#8217;s disease is a neurodegenerative disorder that results from the degeneration of dopaminergic neurons in the central nervous system, ...

3D Kinematic Analysis for the Functional Evaluation in the Rat Model of Sciatic Nerve Crush Injury

坐骨神经粉碎损伤大鼠模型中功能评估的3D运动学分析

Compared to the Sciatic Functional Index (SFI), kinematic analysis is a more reliable and sensitive method for performing functional evaluations of ...

Semi-Quantitative Determination of Dopaminergic Neuron Density in the Substantia Nigra of Rodent Models using Automated Image Analysis

使用自动图像分析对啮齿动物模型的亚斯坦蒂娅尼格拉多巴明神经元密度进行半定量测定

Estimation of the number of dopaminergic neurons in the substantia nigra is a key method in pre-clinical Parkinson's disease research. Currently, unbiased ...

A Rat Model of EcoHIV Brain Infection

生态HIV脑感染的老鼠模型

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. ...

Motor Dual-Tasks for Gait Analysis and Evaluation in Post-Stroke Patients

用于中风后患者步态分析和评估的运动双任务

Eighteen stroke patients were recruited for this study involving the evaluation of cognition and walking ability and multitask gait analysis. Multitask ...

The 6-hydroxydopamine Rat Model of Parkinson's Disease

帕金森病的6-羟基多巴胺大鼠模型

Motor symptoms of Parkinson's disease (PD)-bradykinesia, akinesia, and tremor at rest-are consequences of the neurodegeneration of dopaminergic neurons in ...

Induction and Assessment of Levodopa-induced Dyskinesias in a Rat Model of Parkinson's Disease

左旋多巴诱导的帕金森病大鼠模型中运动障碍的诱导与评估

Levodopa (L-DOPA) remains the gold-standard therapy used to treat Parkinson&#39;s disease (PD) motor symptoms. However, unwanted involuntary movements ...