Name: low-cost gait analysis for behavioral phenotyping of mouse models of neuromuscular disease
Uploaded: 2019-07-18T15:00:00
Description: Watch this Scientific Journal Video about Low-Cost Gait Analysis for Behavioral Phenotyping of Mouse Models of Neuromuscular Disease at JoVE.com

作者感谢A.M.提供的动物鉴定援助。这项工作得到了美国国家卫生研究院(R01 7 RF1 AG057264 至 A.R.L.S.和 C.J.C.和 R01 NS100023 到 A.R.L.S) 和肌肉萎缩协会(基础研究补助金给 A.R.L.S.S.,发展赠款给 C.J.C.S.)的资助。

杜克大学机构动物护理和使用委员会(IACUC)对小鼠进行的所有测试都经过审查和批准。负责测试和评分的人员必须对动物基因型或实验条件视而不见,直到整个队列完成步态分析和论文评分。
1. 测试材料准备
<ol><li>使用 3 块厚度为 0.375 英寸的预切割透明丙烯酸面板建造的隧道进行测试。将面板与密封剂结合在一起组装隧道,这种密封剂专门粘结丙烯酸,干燥时不会散发出异味。<...

<ol>
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Q., 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=RNA+interference+improves+motor+and+neuropathological+abnormalities+in+a+Huntington's+disease+mouse+model.">RNA interference improves motor and neuropathological abnormalities in a Huntington's disease mouse model.</a> Proceedings of the National Academy of Sciences of the United States of America. 102 (16), 5820-5825 (2005).</li><li>Lin, C. H., 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=Neurological+abnormalities+in+a+knock-in+mouse+model+of+Huntington's+disease.">Neurological abnormalities in a knock-in mouse model of Huntington's disease.</a> Human Molecular Genetics. 10 (2), 137-144 (2001).</li><li>Sopher, B. L., 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=Androgen+receptor+YAC+transgenic+mice+recapitulate+SBMA+motor+neuronopathy+and+implicate+VEGF164+in+the+motor+neuron+degeneration.">Androgen receptor YAC transgenic mice recapitulate SBMA motor neuronopathy and implicate VEGF164 in the motor neuron degeneration.</a> Neuron. 41 (5), 687-699 (2004).</li><li>Tillerson, J. L., Caudle, W. M., Reveron, M. E., Miller, G. 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采用上述低成本步态分析方法,在SBMA的BAC fxAR121小鼠模型中,成功地识别了症状后年龄的步态功能障碍的几个参数。步幅长度的减小与先前SBMA对小鼠模型和人类患者的研究一致。我们还首次表明,与非转基因的杂物对照相比,有症状的SBMA小鼠的后肢脚趾扩散存在显著差异。有趣的是,后脚趾扩张的减少可能是由爪子扩张肌肉的虚弱、爪子屈肌的紧绷或神经内侧不良2,19引起,这也与SBMA的...

运动需要复杂的神经和肌肉骨骼协调,在运动路径的单一方面出现缺陷可以产生可观察到的步态异常1,2。盖特分析是研究人员测试小鼠模型的关键工具,因为它提供了可量化的行为数据,了解特定疾病、损伤或药物如何影响动物的运动3。然而,数字化步态分析需要购买跑步机、照相机和相关软件,这对研究人员来说可能非常昂贵。盖特分析经常间歇性地用于跟踪电机功能的纵向变化,因此,如果偶尔使用4,则很难证明支出的合理性。尽管数字化分析可能比简单的足迹分析提供更详细的步态指标,但这些更复杂的度量并不总是必要的,也并不总是与行为表型5的表征相关。
在这里,我们提出了一个低成本的手动足迹分析方法,作为一个快速和敏感的替代数字化步态分析程序6,7。人工足迹分析已被证明,以检测在众多的鼠病模型4,7,8,9,10,11的显著步态差异 ,12,13,14,15,16,17,和至少一个情况下, 这种低成本的方法识别步态的变化,没有检测到一个常见的数字化步态分析程序12。材料的总成本是象征性的,它可以很容易地适应其他啮齿动物研究模型。
虽然可以从中绘制数据,但我们描述的方法侧重于三个特定的指标:步长、步长宽度(又名"轨道宽度")和脚趾分布。必须指出,应逐个模型确定要评估的参数。这种步态分析方法不是用来测量认知功能的,也不推荐用于需要复杂的步态生物力学测量的研究。
我们提供来自一组症状前和症状后小鼠的行为数据,这些小鼠模拟X链接脊柱和布尔巴肌肉萎缩症(SBMA),这是一种神经肌肉疾病,其特征是运动神经元退化和肌肉萎缩。这些小鼠在步态中出现渐进性缺陷,与其他疾病特异性表型的发病一致。这证明了这种方法的有效性和特异性,并证实它可以可靠地区分受影响的和未受影响的动物。
本研究中的实验鼠为2.5岁(症状前)和9个月大(症状后)BAC fxAR121转基因小鼠,背景为C57BL/6(nexpt=12)。该模型在我们的实验室生成,并已被完全描述为SBMA9的强大鼠标模型。非转基因杂物用作对照(nctrl#8)。SBMA是一种性别限制疾病,只在雄性中完全显现出来,因此雄性小鼠被专门用于这项研究。在规划阶段,研究人员必须考虑到国家卫生研究院对性别作为生物变量的考虑,以确定群体的大小和组成18。

<table>
	<tbody>
		<tr>
			<td>Caliper</td>
			<td>n/a</td>
			<td>n/a</td>
			<td>must have markings down to 0.1 mm</td>
		</tr>
		<tr>
			<td>Craft Glue</td>
			<td>E6000</td>
			<td>n/a</td>
		</tr>
		<tr>
			<td>Footprint Paint (Tempera Paint)</td>
			<td>Artmind</td>
			<td>n/a</td>
			<td>must be non-toxic</td>
		</tr>
		<tr>
			<td>Round Barrel Paintbrushes</td>
			<td>Symply Simmons</td>
			<td>n/a</td>
			<td>0.5 cm diameter</td>
		</tr>
		<tr>
			<td>Ruler</td>
			<td>n/a</td>
			<td>n/a</td>
			<td>must have markings down to millimeters</td>
		</tr>
		<tr>
			<td>Scoring Paper (Watercolor Pads)</td>
			<td>Canson</td>
			<td>n/a</td>
			<td>cut to size</td>
		</tr>
		<tr>
			<td>Tunnel and Goal Chamber</td>
			<td>Interstate Plastics</td>
			<td>n/a</td>
			<td>cut to size</td>
		</tr>
	</tbody>
</table>

low-cost gait analysis for behavioral phenotyping of mouse models of neuromuscular disease

动物运动测量是一种常见的行为工具,用于描述特定疾病、损伤或药物模型的表型。这里所展示的低成本步态分析方法是一种简单但有效的测量鼠步态异常的方法。通过用无毒的可洗漆涂一个老鼠的脚,让受试者在一张纸上穿过隧道来分析脚印。测试隧道的设计利用了自然小鼠行为及其对小黑暗场所的亲和力。使用尺子和铅笔可以轻松测量每只鼠标的步长、步幅宽度和脚趾分布。这是一种成熟且可靠的方法,它生成多个类似于数字系统的指标。这种方法足够灵敏,可以检测表型表示的早期步幅变化,并且由于其非侵入性方法,它允许在寿命或表型表示中测试组。

由于动物数量充足,这个程序能够检测小鼠基因型之间的步态差异,随着时间的推移,在相同的应变内。图1B显示了在我们的实验室收集的足迹图像的代表性痕迹,使用X链接脊柱和布尔巴肌肉萎缩(SBMA)的小鼠模型,这是一种影响下部运动神经元和骨骼肌的神经退行性疾病。我们以前曾报道过,雄性BACfxAR121转基因小鼠与非转基因杂物对照组相比,在症状后年龄出现显?...

Watch this Scientific Journal Video about Low-Cost Gait Analysis for Behavioral Phenotyping of Mouse Models of Neuromuscular Disease at JoVE.com

Low-Cost Gait Analysis for Behavioral Phenotyping of Mouse Models of Neuromuscular Disease

足迹分析是数字化步态分析程序的低成本替代方案,用于研究人员量化小鼠的运动异常。由于其速度、简单性和纵向潜力,它是小鼠模型行为型型的理想选择。

神经肌肉疾病小鼠模型行为花型的低成本盖特分析

Measurement of animal locomotion is a common behavioral tool used to describe the phenotype of a given disease, injury, or drug model. The low-cost method ...

足迹分析是一种低成本的替代方案，用于数字化步态分析程序，用于研究人员量化小鼠的运动异常。由于其速度、简单性和纵向潜能，此方法非常适合小鼠模型的行为表型，包括神经退行性疾病、神经肌肉疾病或中风模型。演示这个程序的将是弗吉尼亚·韦特曼，一个来自我实验室的研究技术员。在开始实验之前，让完全清醒并警觉至少5分钟的小鼠适应检测室30分钟。将隧道放置在略宽、比隧道长度和宽度长的纸条上。使用鼠标 ID 标记纸张和测试日期。根据需要将向日葵种子添加到目标室，以激励，并牢固地将第一只要测试的小鼠擦伤。使用直径约 0.5 厘米的锥形刷尖，用一种无毒可洗水性油漆颜色绘制所有脚趾的整个底面和每个前爪的中心。以类似的方式涂上对比鲜明的无毒可洗水性油漆颜色的后爪。使用干净的湿布去除鼠标在其身体其他部位上得到的任何油漆，以防止可能干扰数据收集的污迹。然后将鼠标放在隧道的开始处。让鼠标一直进入目标室，然后取回动物用水阻尼布轻轻清洁其脚。然后将鼠标返回到其家庭笼子，并在测试下一只动物之前用消毒剂擦拭测试区域和隧道。为进行准确评分，在选择具有透明无污迹的脚印的步数之前，允许尺寸完全干燥。要生成足够的评分数据，每个英尺必须至少连续两个步骤。不要在纸上包含第一个和最后一个脚印，因为它们不太可能代表正常的步态，因为鼠标正在改变其行走速度。要将步幅长度定义为同一英尺创建的两个连续脚印之间的距离，请使用铅笔在单个步幅中围绕两个前肢脚印前脚区域绘制两到四毫米的圆，并使用标尺在圆之间绘制一条线。将每个圆中间的两个打印件之间的距离记录为右前一或左前一个，并根据需要对可评分的每个步骤重复测量。然后，平均所有个体记录的步幅距离，每个肢体平均单个分数在一组一起作为实验适当。要将步幅宽度定义为左前肢或后肢之间的距离，请从一个后肢的圆形前脚区域绘制并测量一条线，该线与反侧后肢上步幅长度的线垂直相交。对可评分的所有后印打印重复此测量，并平均测量值。要将脚趾张差定义为单个前肢或后刻脚印上的第一个脚趾和最后一个脚趾之间的距离，请使用卡钳测量第一个脚趾打印的尖端和最后一个脚趾打印的尖端之间的距离。然后重复所有可评分的后印打印的测量，并平均测量值。对于足够数量的动物，这个程序能够检测小鼠基因型在相同菌株中随着时间的推移的步态差异。例如，这里显示了使用 x 链接脊髓和球茎肌肉萎缩的小鼠模型收集的足迹图像的代表性痕迹，一种影响下运动神经元和骨骼肌的神经退行性疾病。对症状前和症状后转基因和垃圾伴侣对照的盖特分析表明，在疾病发生之前，转基因小鼠的步幅、步幅和足趾传播与它们的垃圾伴侣非转基因对照相似。然而，在疾病发病后，转基因动物的步幅明显变短。类似的纵向分析显示，在测试的两个年龄段宽度没有差异。症状后转基因小鼠的后脚趾传播也明显较年龄匹配的垃圾伴侣对等。将油漆正确涂在爪子上将产生最佳效果。请记住，只有始终带透明无污迹脚印的步骤才应评分。此技术足够灵敏，可以检测步幅的细微变化。由于其非侵入性方法，它允许测试在其生命周期中组表型呈现和治疗干预。

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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

Nest Building as an Indicator of Health and Welfare in Laboratory Mice

The minimization and alleviation of  suffering has moral and scientific implications. In order to mitigate this  negative experience one must be able to ...

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 ...

Comprehensive Analysis of Transcription Dynamics from Brain Samples Following Behavioral Experience

转录动力学的脑组织样本在行为经验综合分析

The encoding of experiences in the brain and the consolidation of long-term memories depend on gene transcription. Identifying the function of specific ...

The Treadmill Fatigue Test: A Simple, High-throughput Assay of Fatigue-like Behavior for the Mouse

跑步机疲劳试验：疲劳类似的行为，为鼠标操作简单，高通量检测

Fatigue is a prominent symptom in many diseases and disorders and reduces quality of life for many people. The lack of clear pathogenesis and failure of ...

A Simple and Low-cost Assay for Measuring Ambulation in Mouse Models of Muscular Dystrophy

一种简便、低成本的肌营养不良小鼠步行模型的测定方法

Measuring functional outcomes in the treatment of muscular dystrophy is an essential aspect of preclinical testing. The assessment of voluntary ambulation ...

Clinical-oriented Three-dimensional Gait Analysis Method for Evaluating Gait Disorder

基于临床的三维步态分析方法评价步态障碍

Three-dimensional gait analysis (3DGA) is shown to be a useful clinical tool for the evaluation of gait abnormality due to movement disorders. 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 ...

Low-cost Protocol of Footprint Analysis and Hanging Box Test for Mice Applied the Chronic Restraint Stress

小鼠应用慢性约束应激的低成本足迹分析和吊箱试验协议

Gait disturbance is frequently observed in patients with movement disorders. In mouse models used for movement disorders, gait analysis is important ...

Paw-Print Analysis of Contrast-Enhanced Recordings (PrAnCER): A Low-Cost, Open-Access Automated Gait Analysis System for Assessing Motor Deficits

Paw-Print Analysis of Contrast-Enhanced Recordings PrAnCER: A Low-Cost, Open-Access Automated Gait Analysis System for Assessing Motor Deficits

对比度增强记录 (PrAnCER) 的爪打印分析:用于评估电机缺陷的低成本、开放访问自动步态分析系统

Gait analysis is used to quantify changes in motor function in many rodent models of disease. Despite the importance of assessing gait and motor function ...