Name: automated gait analysis in mice with chronic constriction injury
Uploaded: 2017-10-17T14:00:00
Description: Watch this Scientific Journal Video about Automated Gait Analysis in Mice with Chronic Constriction Injury at JoVE.com

这项研究得到了南国立大学、韩国东方医学研究所 (KIOM) 的支持, 并通过韩国卫生产业发展研究所 (KHIDI) 资助了韩国卫生技术研发 #38;D 项目, 由卫生部提供资金。和 #38; 大韩民国福利 (赠款号: HI15C0007)。

所有实验都是按照国际疼痛研究协会的道德准则进行的, 并得到了南国立大学机构动物保育和使用委员会的批准 (韩国大田). 
 1. 在坐骨神经上的诱导作用 <ol> <li> 家庭男性民事代表小鼠在受控室温下的 12 h 光照/暗循环下重 20-25 克 (维持在 20-25 和 #176; C) 和湿度 (40-60%), 可免费获得食物和水.在手术前至少1周, 允许动物保育室的小鼠适应时间. 
	<ol> <li> 在外壳期间,...

<ol>
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P., Bories, C., De Koninck, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simplified+up-down+method+(SUDO)+for+measuring+mechanical+nociception+in+rodents+using+von+Frey+filaments.">A simplified up-down method (SUDO) for measuring mechanical nociception in rodents using von Frey filaments.</a> Mol Pain. 10, 26 (2014).</li><li>Chaplan, S. R., Bach, F. W., Pogrel, J. W., Chung, J. M., Yaksh, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+assessment+of+tactile+allodynia+in+the+rat+paw.">Quantitative assessment of tactile allodynia in the rat paw.</a> J Neurosci Methods. 53 (1), 55-63 (1994).</li><li>Chen, H., Du, J., Zhang, Y., Barnes, K., Jia, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishing+a+Reliable+Gait+Evaluation+Method+for+Rodent+Studies.">Establishing a Reliable Gait Evaluation Method for Rodent Studies.</a> J Neurosci Methods. , (2017).</li><li>Kang, D. W., 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=Antinociceptive+Profile+of+Levo-tetrahydropalmatine+in+Acute+and+Chronic+Pain+Mice+Models:+Role+of+spinal+sigma-1+receptor.">Antinociceptive Profile of Levo-tetrahydropalmatine in Acute and Chronic Pain Mice Models: Role of spinal sigma-1 receptor.</a> Sci Rep. 6, 37850 (2016).</li><li>Huehnchen, P., Boehmerle, W., Endres, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+paclitaxel+induced+sensory+polyneuropathy+with+&amp;#34;Catwalk&amp;#34;+automated+gait+analysis+in+mice.">Assessment of paclitaxel induced sensory polyneuropathy with &amp;#34;Catwalk&amp;#34; automated gait analysis in mice.</a> PLoS One. 8 (10), e76772 (2013).</li><li>Vrinten, D. H., Hamers, F. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term='CatWalk'+automated+quantitative+gait+analysis+as+a+novel+method+to+assess+mechanical+allodynia+in+the+rat;+a+comparison+with+von+Frey+testing.">'CatWalk' automated quantitative gait analysis as a novel method to assess mechanical allodynia in the rat; a comparison with von Frey testing.</a> Pain. 102 (1-2), 203-209 (2003).</li><li>Martinov, T., Mack, M., Sykes, A., Chatterjea, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+changes+in+tactile+sensitivity+in+the+hind+paw+of+mice+using+an+electronic+von+Frey+apparatus.">Measuring changes in tactile sensitivity in the hind paw of mice using an electronic von Frey apparatus.</a> J Vis Exp. (82), e51212 (2013).</li><li>Ferland, C. E., Laverty, S., Beaudry, F., Vachon, P. <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+and+pain+response+of+two+rodent+models+of+osteoarthritis.">Gait analysis and pain response of two rodent models of osteoarthritis.</a> Pharmacol Biochem Behav. 97 (3), 603-610 (2011).</li><li>Mogil, J. 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=Hypolocomotion,+asymmetrically+directed+behaviors+(licking,+lifting,+flinching,+and+shaking)+and+dynamic+weight+bearing+(gait)+changes+are+not+measures+of+neuropathic+pain+in+mice.">Hypolocomotion, asymmetrically directed behaviors (licking, lifting, flinching, and shaking) and dynamic weight bearing (gait) changes are not measures of neuropathic pain in mice.</a> Mol Pain. 6, 34 (2010).</li><li>Ferreira-Gomes, J., Adaes, S., Castro-Lopes, J. 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目前, 在疼痛动物模型中, 用痛纤维测量机械性的方法是最常用的方式来表现触觉过敏。随着神经性疼痛动物模型的不断发展, 感官功能评估的方法也得到了改善8,9,10,15。在这些报告中, 有人建议这些修改可以为旧方法提供更准确、快速和 user-friendly 的替换。然而, 这些方法的数据仍然可能受到其主观性...

创伤、代谢功能障碍、炎症、感染、缺血或自身免疫性疾病引起的神经系统病理改变, 有时会导致神经病理性疼痛, 这被定义为损害或疾病的直接后果所引起的疼痛。影响体感系统1。神经病理性疼痛通常是无法忍受的, 不幸的是, 常规镇痛药一般不会产生足够的疼痛缓解2。神经病理性疼痛的一个主要特点是自发的和刺激诱发的 (即, 痛和痛觉过敏) 疼痛。痛是一种伤害反应, 发生在通常非刺激, 如轻触或温暖刺激。痛觉过敏表示对有害的机械和/或热刺激的疼痛反应增强3。尽管这两种症状都严重损害了患者的生活质量, 但轻柔的触觉刺激所诱发的机械痛是最严重的症状, 因为在日常生活中很难避免接触。
为了探讨治疗神经病理性疼痛的基本机制和新的镇痛药, 对疼痛反应的精确测量是必不可少的。许多神经病理性疼痛动物模型已经发展到后爪区的伤害反应, 因为它的高可及性4,5,6,7。因此, 大多数疼痛反应评估已经执行的脚底或背部后爪表面的应用机械刺激使用特殊仪器, 如弗雷丝。最常用的方法之一是狄克逊8和以后修改后的版本910中描述的向上和向下的方法。然而, 非常熟练, 经验丰富的实验者需要执行冯弗雷测试, 结果可能是主观的。
自动步态分析系统可以通过测量在自由移动的啮齿动物中行走的各种参数来研究神经和神经肌肉紊乱。在各种神经损伤动物模型中, 伤害的程度和几种治疗方法的镇痛效果可以在不增加疼痛刺激的情况下进行评估11,12,13,14. 该分析系统可检测静态和动态步态参数, 如: 爪印区域 (与地板接触的完整爪印区域)、爪子强度 (接触爪面积的平均强度)、步幅长度 (同一个爪子的连续安置之间的距离), 姿态阶段 (地面接触的持续时间为一只后腿), 步顺序 (命令四爪被安置在地板上), 摇摆 (摇摆阶段的期间) 和摇摆速度 (计算从步长和摆动持续时间, 以每秒的像素表示)。本文介绍了使用一个分析系统, 并提供了一个简单的数据与冯弗雷试验使用慢性收缩损伤 (局) 神经病理性小鼠。

<table>
	<tbody>
		<tr>
			<td>0.9% Saline</td>
			<td>JW Pharmaceutical</td>
			<td>N/A</td>
			<td>Vehicle for drugs</td>
		</tr>
		<tr>
			<td>1ml syringe</td>
			<td>BD Plastipak</td>
			<td>300013</td>
			<td>Injecting device</td>
		</tr>
		<tr>
			<td>2, 2, 2-tribromoethanol (97% purity)</td>
			<td>Sigma</td>
			<td>T48402</td>
			<td>Anesthetic</td>
		</tr>
		<tr>
			<td>2-methyl-2-buthanol (99% purity)</td>
			<td>Sigma</td>
			<td>152463</td>
			<td>Solvent for 2, 2, 2-tribromoethanol</td>
		</tr>
		<tr>
			<td>Catwalk Automated gait analysis system</td>
			<td>Noldus</td>
			<td>N/A</td>
			<td>Automatic analysis software of aniaml gait</td>
		</tr>
		<tr>
			<td>Chromic catgut (4-0 thickness)</td>
			<td>AILEE</td>
			<td>C442</td>
			<td>Ligature to make chronic constriction injury on the sciatic nerve</td>
		</tr>
		<tr>
			<td>Gabapentin</td>
			<td>Sigma</td>
			<td>Y0001280</td>
			<td>Analgeisc, Used as a positive control drug in this study</td>
		</tr>
		<tr>
			<td>Graefe Forceps</td>
			<td>F.S.T</td>
			<td>11051-10</td>
			<td>Surgical instrument</td>
		</tr>
		<tr>
			<td>Heating Pad</td>
			<td>DAESHIN ELECTRONICS</td>
			<td>M-303AT</td>
			<td>Regulation of body temperature</td>
		</tr>
		<tr>
			<td>ICR Mouse</td>
			<td>Samtaco</td>
			<td>N/A</td>
			<td>Experimental animal</td>
		</tr>
		<tr>
			<td>Mersilk (3-0 thickness)</td>
			<td>ETHICON</td>
			<td>W598H</td>
			<td>Suture material for surgical closure of skin</td>
		</tr>
		<tr>
			<td>Micro-Mosquito</td>
			<td>F.S.T</td>
			<td>13010-12</td>
			<td>Surgical instrument</td>
		</tr>
		<tr>
			<td>Micro-scissors</td>
			<td>F.S.T</td>
			<td>14090-09</td>
			<td>Surgical instrument</td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>F.S.T</td>
			<td>12002-12</td>
			<td>Surgical instrument</td>
		</tr>
		<tr>
			<td>Povidone Iodine</td>
			<td>Firson</td>
			<td>N/A</td>
			<td>Disinfectant to prevent infection after surgery</td>
		</tr>
		<tr>
			<td>Scalpel blade</td>
			<td>F.S.T</td>
			<td>10010-00 (#10)</td>
			<td>Surgical instrument to make an incision</td>
		</tr>
		<tr>
			<td>Scalpel handle</td>
			<td>F.S.T</td>
			<td>10003-12 (#3)</td>
			<td>Surgical instrument to make an incision</td>
		</tr>
		<tr>
			<td>Von-Frey filaments</td>
			<td>North Coast</td>
			<td>NC12775-99</td>
			<td>Measurement device to test sensory function for mechanical stimulation</td>
		</tr>
	</tbody>
</table>

automated gait analysis in mice with chronic constriction injury

弗雷试验是一种被广泛用于检测神经病理性疼痛动物感官功能的经典方法。然而, 它也有一些缺点, 如主观数据和要求的熟练, 经验丰富的实验者。到目前为止, 各种修改改进了冯弗雷方法, 但它仍然有一些限制。最近的报告表明, 步态分析产生更准确和客观的数据从神经病动物。该协议演示了如何执行自动步态分析, 以确定的神经病理性疼痛的小鼠。经过几天的驯化, 老鼠被允许在玻璃地板上自由行走以照亮脚印。然后, 通过对各种步行参数的自动分析, 如爪印面积、摆动时间、爪角、等, 对脚印和步态进行量化。
本研究的主要目的是描述自动步态分析的方法, 并将其与经典感官试验中的数据进行简单比较。

我们已经执行了冯弗雷测试和自动步态分析在会展小鼠, 直到10天后的手术。对于统计分析, two-way 方差分析的重复测量方法 (方差) 决定了总体效果, 并对 Dunnett 的事后进行了分析, 以确定实验组之间的 p 值。
图 1所示的结果表明了经典的冯弗雷测试数据的时间过程。与假组相比, PWF (%) 小鼠在术后2天...

Watch this Scientific Journal Video about Automated Gait Analysis in Mice with Chronic Constriction Injury at JoVE.com

Automated Gait Analysis in Mice with Chronic Constriction Injury

准确评估神经病理性动物模型的疼痛反应, 对于研究疼痛疾病的病理生理学和开发新的镇痛药是至关重要的。我们提出了一个敏感和客观的方法, 以确定的感官功能的啮齿动物后爪的自动步态分析系统。

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Peripheral and central nerve injuries are mostly studied in rodents, especially rats, given the fact that these animal models are both cost-effective and ...

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

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

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

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Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and ...