这项工作是支持由美国国立卫生研究院拨款NS052535聚糖EY014061＆EY014997的ARL，以及UL1DE019583 - 02财团奖。

为了避免偏见，实验者进行的评估不应该有动物的基因型知识。个别措施，取得了0-3分，0代表没有相关的表型和代表的最严重的表现的3。每个测试进行多次，以确保将比分是可重复的的。肥胖会所述的所有措施的解释复杂化。研究者不妨权衡表型的得分，以评估的结果可能发挥的作用，肥胖​​的老鼠。 礁测试窗台测试是一个直接的措施，协调，这是在小脑共济失调和许多其他神经退行性疾病中受损。这项措施是最直接可比人类小脑共济失调的迹象。 <ol><li>电梯从笼子里的鼠标和它放在笼子里的窗台上。老鼠通常会沿着窗台，企图入笼下降。 </li><li>观察鼠标，因为它走笼窗台，并降低其笼。野生型的小鼠通常会沿着窗台，而不会失去其平衡，会降低入笼优雅，用它的爪子。这是分配的得分为0。如果鼠标失去其立足点而沿窗台走，但在其他方面出现的协调，它接收到1分。如果没有有效地利用它的后腿，发挥得淋漓尽致，而不是它的爪子或土地入笼下降时，它接收到2分。如果脱落的窗台，或接近，而步行或试图降低自己，或动摇，并拒绝移动尽管鼓励所有，它接收到3分。有些老鼠会需要一个温柔的微调，以鼓励他们沿着窗台，或入笼下降。 </li><li>记录窗台测试得分。 </li></ol>后肢紧握后肢紧握的病情恶化，在小鼠模型的神经退行性疾病，包括某些小脑共济失调[1]的标记。 <ol><li>抓住它的基地附近的尾巴，并解除所有周围的物体清晰的鼠标。 </li><li>观察10秒的后肢位置。如果坚持后肢叉开向外，从腹部，它是分配的得分为0。如果一个后肢是朝着腹部的暂停时间超过50％的翻供，它接收到1分。如果两个后肢部分收回对暂停的时间超过50％的腹部时，它接收一个2分。如果它的后肢完全缩回，触摸腹部的暂停时间超过50％，它接收一个3分。 </li><li>将鼠标放回笼子里，并记录其后肢紧握得分。 </li></ol>步态步态是一个协调和肌肉功能的措施。 <ol><li>鼠标从笼中取出，并放置在一个平坦的表面，其头部远离研究者面临。 </li><li>观察鼠标，从后面行走。如果鼠标移动正常，支持其体重四肢，腹部不接触地面，并与后肢均参加均匀，收到了0分。如果它显示了震颤，或出现跛行，行走时，它接收1分。如果它显示了剧烈震颤，严重跛行，降低骨盆，或从身体，脚点在运动（“鸭脚”），它接收到2分。如果鼠标有困难向前迈进，并拖动它的腹部，沿地面的，它接收一个3分。 </li><li>将鼠标放回笼子里，并记录其步态评分。 </li></ol>脊柱后凸后凸畸形的一个特点是背的脊柱弯曲，常见的表现是神经退行性疾病小鼠模型[2]。这是由于继发于神经退行性病变的脊髓肌肉中的肌张力损失。 <ol><li>鼠标从笼中取出，放置在一个平面上上。 </li><li>观察它，因为它走。如果鼠标是能够很容易地拉直脊柱，因为它散步，不具有持久性脊柱后凸的，它接收得分为0。如果鼠标表现出轻度的脊柱后凸畸形，但能够理顺其脊椎，它接收到1分。如果不能彻底整顿其脊椎，并保持持续，但轻度后凸，它接收到2分。如果鼠标保持着明显的后凸畸形，如散步或而坐，它被分配一个3分。 </li><li>将鼠标放回笼子里，并记录其后凸得分。 </li></ol>代表性的成果有了足够数量的动物，这个程序能够检测菌株间内随着时间的推移一脉相承的表型差异。进行数据分析，计算所采取的每项措施的得分平均每一个人的三个测量。措施可以单独分析，或结合成复合材料的表型得分为各个。 图表中的数据和应用适当的统计方法来确定的意义。图1显示了从转基因小鼠SCA7模型的复合的表型评估的结果。在此floxed SCA7 - 92Q的转基因模型，两侧是人类ataxin - 7基因与92个CAG重复loxP位的网站，并从细菌人工染色体表达。复合材料的表型得分的紧握着，窗台走路，步态最大可能得分12和后凸评估。逐步SCA7在floxed SCA7 - 92Q的转基因小鼠的表型表现出越来越大的复合表型的得分，这与人类疾病的逐步性质是一致的。 <img src="/files/ftp_upload/1787/1787fig1.jpg" alt="图1" /> 图1 Floxed SCA7 92Q小鼠表现出一个渐进的SCA7表型，即开始显着不同的非转基因窝，在20周（2方差分析 ：Bonferroni事后; *** P &lt; 0.001）。一个窗台测试每0-3规模的小鼠进行了评估，紧握着，步态和后凸畸形。每个基因型的平均综合得分，每个年龄计算。酒吧代表教育统筹局局长。

<ol>
	<li>Chou, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polyglutamine-expanded+ataxin-3+causes+cerebellar+dysfunction+of+SCA3+transgenic+mice+by+inducing+transcriptional+dysregulation.">Polyglutamine-expanded ataxin-3 causes cerebellar dysfunction of SCA3 transgenic mice by inducing transcriptional dysregulation.</a> Neurobiol Dis. 31 (1), 89-101 (2008).</li><li>Thomas, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Loss+of+endogenous+androgen+receptor+protein+accelerates+motor+neuron+degeneration+and+accentuates+androgen+insensitivity+in+a+mouse+model+of+X-linked+spinal+and+bulbar+muscular+atrophy.">Loss of endogenous androgen receptor protein accelerates motor neuron degeneration and accentuates androgen insensitivity in a mouse model of X-linked spinal and bulbar muscular atrophy.</a> Hum Mol Genet. 15 (14), 2225-2238 (2006).</li><li>Ditzler, S., Stoeck, J., LeBlanc, M., Kooperberg, C., Hansen, S., Coppin, L., Olson, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Rapid+Neurobehavioral+Assessment+Reveals+that+FK506+Delays+Symptom+Onset+in+R6/2+Huntington's+Disease+Mice.">A Rapid Neurobehavioral Assessment Reveals that FK506 Delays Symptom Onset in R6/2 Huntington's Disease Mice.</a> Preclinica Research Articles. 1 (3), 115-126 (2003).</li></ol>

这些研究的大学华盛顿机构动物护理和使用委员会批准。

此协议是专为小脑共济失调的小鼠模型中的疾病严重程度的一个敏感和容易执行的评价。的打分系统的各个组件将在不同的神经退行性疾病小鼠模型或多或少有效。 这个评分系统的元素都被有效地用于评估的各种人类神经退行性疾病的小鼠模型，包括小脑共济失调，亨廷顿氏病和spinobulbar性肌萎缩症[1-3]。测试的理想组合将取决于问题的障碍。该协议最初是由作者设计，评估?...

a simple composite phenotype scoring system for evaluating mouse models of cerebellar ataxia

我们描述了一种快速，灵敏的定量小脑共济失调的小鼠模型中的疾病严重程度的协议。这是来自先前公布的表型在一些疾病模型评估，包括脊髓小脑共济失调，亨廷顿氏病和spinobulbar肌肉萎缩。措施包括后肢紧握着，窗台测试，步态和后凸。记录每一个措施是0-3分，与0-12的所有四项措施的总和。结果有效区分影响和非受影响的个人，同时也量化的神经退行性疾病表型的时间进展。措施可以单独分析，或结合成一个更大的统计力量的综合表型得分。理想的组合描述的四个措施将取决于问题的障碍。我们目前用来评估疾病的严重程度在脊髓小脑性共济失调7型（SCA7）的转基因小鼠模型的协议的一个例子。 阿尔伯特R.拉斯帕达和Gwenn A.花园贡献同样这个手稿。

Watch this Scientific Journal Video about A Simple Composite Phenotype Scoring System for Evaluating Mouse Models of Cerebellar Ataxia at JoVE.com

A Simple Composite Phenotype Scoring System for Evaluating Mouse Models of Cerebellar Ataxia

我们描述了一种快速，灵敏的定量小脑共济失调的小鼠模型中的疾病严重程度的协议。措施包括后肢紧握着，窗台测试，步态和后凸。该协议有效地区别对待受影响的和非受影响的个人，以及检测，随着时间的推移受影响的个人的进展。

一个简单的综合评分系统评估小脑共济失调的小鼠模型的表型

We describe a protocol for the rapid and sensitive quantification of disease severity in mouse models of cerebella ataxia. It is derived from previously ...

a-simple-composite-phenotype-scoring-system-for-evaluating-mouse

Research

JoVE Journal

Neuroscience

35.4K 次观看.  University of Washington. 我们描述了一种快速，灵敏的定量小脑共济失调的小鼠模型中的疾病严重程度的协议。措施包括后肢紧握着，窗台测试，步态和后凸。该协议有效地区别对待受影响的和非受影响的个人，以及检测，随着时间的推移受影响的个人的进展。

视频: A Simple Composite Phenotype Scoring System for Evaluating Mouse Models of Cerebellar Ataxia

Clinical Testing and Spinal Cord Removal in a Mouse Model for Amyotrophic Lateral Sclerosis (ALS)

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder resulting in progressive degeneration of motoneurons. Peak of onset is around 60 years for the sporadic disease and around 50 years for the familial disease. Due to its progressive course, 50% of the patients die within 30 months of symptom onset. In order to evaluate novel treatment options for this disease, genetic mouse models of ALS have been generated based on human familial mutations in the SOD gene, such as the SOD1 (G93A) mutation. Most important aspects that have to be evaluated in the model are overall survival, clinical course and motor function. Here, we demonstrate the clinical evaluation, show the conduction of two behavioural motor tests and provide quantitative scoring systems for all parameters. Because an in depth analysis of the ALS mouse model usually requires an immunohistochemical examination of the spinal cord, we demonstrate its preparation in detail applying the dorsal laminectomy method. Exemplary histological findings are demonstrated. The comprehensive application of the depicted examination methods in studies on the mouse model of ALS will enable the researcher to reliably test future therapeutic options which can provide a basis for later human clinical trials.

Clinical Testing and Spinal Cord Removal in a Mouse Model for Amyotrophic Lateral Sclerosis ALS

A mouse model for amyotrophic lateral sclerosis (ALS) is examined clinically and behaviorally. As a prerequisite for an accompanying immunohistological analysis the preparation of the spinal cord is depicted in detail.

临床试验和脊髓去除在肌萎缩性侧索硬化症的小鼠模型（ALS）

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder resulting in progressive degeneration of motoneurons. Peak of onset is around 60 ...

科研

医学

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

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 of gait analysis demonstrated here is a simple but effective measure of gait abnormalities in murine models. Footprints are analyzed by painting a mouse&#8217;s feet with non-toxic washable paint and allowing the subject to walk through a tunnel on a sheet of paper. The design of the testing tunnel takes advantage of natural mouse behavior and their affinity for small dark places. The stride length, stride width, and toe spread of each mouse is easily measured using a ruler and a pencil. This is a well-established and reliable method, and it generates several metrics that are analogous to digital systems. This approach is sensitive enough to detect changes in stride early in phenotype presentation, and due to its non-invasive approach, it allows for testing of groups across life-span or phenotypic presentation.

Footprint analysis is a low-cost alternative to digitized gait analysis programs for researchers quantifying movement abnormalities in mice. Because of its speed, simplicity, and longitudinal potential, it is ideal for behavioral phenotyping of mouse models.

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 ...

行为学

<meta charset="utf8"></head><body>使用梯形梁测试评估 TH 相关小鼠模型中的运动性能

A Versatile, Behavioral Method to Investigate Thyroid Hormone Effects on Cerebellar Function

Thyroid hormone (TH) action is essential during the development of the central nervous system, including the cerebellum. In case of TH deficiency in early life such as congenital hypothyroidism, patients display neurological disorders such as cognitive retardation and motor deficits. There are various studies using mouse models with tissue- or cell-specific TH deficiency to investigate the role of TH in the cerebellum. Compared to generalized congenital hypothyroid mice, cerebellar cell-specific TH-deficient mice display milder and subtler ataxic features, making the assessment of motor function difficult when using conventional tests such as the rotarod test. 
Due to the need for an alternative tool to assess motor function in TH-related animal models, we developed a versatile behavioral method called the "ladder beam test," in which we can design the various ladder tests depending on the severity of ataxia in model mice. We utilized transgenic mice expressing a dominant-negative TH receptor specifically in the cerebellar Purkinje cell, a sole output neuron in the cerebellar cortex modulating motor performance. The newly-built ladder beam test successfully detected robust impairments in motor performance in the transgenic mice at a greater level compared to the rotarod test. Disruption of motor learning was also detected in the ladder beam test but not in the rotarod test. The protocol with this novel behavioral apparatus can be applied to other animal models that may show mild ataxic phenotype to examine subtle changes in cerebellar function.

<meta charset="utf8"></head><body>作者聚焦:准确评估小鼠甲状腺激素驱动的运动改变

Here we present a protocol for a versatile behavior test developed recently, the ladder beam test. This test has the advantage of detecting subtle cerebellar ataxia caused by a defect of thyroid hormone action in the central nervous system over the conventional behavior tests assessing motor performance.

一种研究甲状腺激素对小脑功能影响的通用行为方法

Thyroid hormone (TH) action is essential during the development of the central nervous system, including the cerebellum. In case of TH deficiency in early ...

使用行为测试验证 Angelman 综合征的小鼠模型

Behavioral Characterization of an Angelman Syndrome Mouse Model

This manuscript describes a battery of behavioral tests available to characterize Angelman syndrome (AS)-like phenotypes in an established murine model of AS. We use the rotarod learning paradigm, detailed gait analysis, and nest building test to detect and characterize animal motor impairments. We test animal emotionality in the open field and elevated plus maze tests, as well as the affect in the tail suspension test. When AS mice are tested in the open field test, the results should be interpreted with care, since motor dysfunctions influence mouse behavior in the maze and alter activity scores.
The reproducibility and effectiveness of the presented behavioral tests has already been validated in several independent Uba3a mouse lines with different knockout variants, establishing this set of tests as an excellent validation tool in AS research. Models with the relevant construct and face validity will warrant further investigations to elucidate the pathophysiology of the disease and grant the development of causal treatments.

作者聚焦:用于验证 Angelman 综合征小鼠模型的一系列高度可重复的行为测试 

This manuscript presents a set of highly reproducible behavioral tests to validate an Angelman syndrome mouse model.

安格曼综合征小鼠模型的行为表征

This manuscript describes a battery of behavioral tests available to characterize Angelman syndrome (AS)-like phenotypes in an established murine model of ...

A Quick Phenotypic Neurological Scoring System for Evaluating Disease Progression in the SOD1-G93A Mouse Model of ALS

The SOD1-G93A transgenic mouse is the most widely used animal model of amyotrophic lateral sclerosis (ALS). At ALS TDI we developed a phenotypic screening protocol, demonstrated in video herein, which reliably assesses the neuromuscular function of SOD1-G93A mice in a quick manner. This protocol encompasses a simple neurological scoring system (NeuroScore) designed to assess hindlimb function. NeuroScore is focused on hindlimb function because hindlimb deficits are the earliest reported neurological sign of disease in SOD1-G93A mice. The protocol developed by ALS TDI provides an unbiased assessment of onset of paresis (slight or partial paralysis), progression and severity of paralysis and it is sensitive enough to identify drug-induced changes in disease progression. In this report, the combination of a detailed manuscript with video minimizes scoring ambiguities and inter-experimenter variability thus allowing for the protocol to be adopted by other laboratories and enabling comparisons between studies taking place at different settings. We believe that this video protocol can serve as an excellent training tool for present and future ALS researchers.

This video protocol describes a sensitive, reliable, and quick method for evaluating the neuromuscular deficits in a transgenic mouse model of amyotrophic lateral sclerosis.

一个快速的表型神经评分系统在ALS的SOD1-G93A小鼠模型评估疾病进展

The SOD1-G93A transgenic mouse is the most widely used animal model of amyotrophic lateral sclerosis (ALS). At ALS TDI we developed a phenotypic screening ...

Treating SCA1 Mice with Water-Soluble Compounds to Non-Specifically Boost Mitochondrial Function

Mitochondrial dysfunction plays a significant role in the aging process and in neurodegenerative diseases including several hereditary spinocerebellar ataxias and other movement disorders marked by progressive degeneration of the cerebellum. The goal of this protocol is to assess mitochondrial dysfunction in Spinocerebellar ataxia type 1 (SCA1) and assess the efficacy of pharmacological targeting of metabolic respiration via the water-soluble compound succinic acid to slow disease progression. This approach is applicable to other cerebellar diseases and can be adapted to a host of water-soluble therapies.
Ex vivo analysis of mitochondrial respiration is used to detect and quantify disease-related changes in mitochondrial function. With genetic evidence (unpublished data) and proteomic evidence of mitochondrial dysfunction in the SCA1 mouse model, we evaluate the efficacy of treatment with the water-soluble metabolic booster succinic acid by dissolving this compound directly into the home cage drinking water. The ability of the drug to pass the blood brain barrier can be deduced using high performance liquid chromatography (HPLC). The efficacy of these compounds can then be tested using multiple behavioral paradigms including the accelerating rotarod, balance beam test and footprint analysis. Cytoarchitectural integrity of the cerebellum can be assessed using immunofluorescence assays that detect Purkinje cell nuclei and Purkinje cell dendrites and soma. These methods are robust techniques for determining mitochondrial dysfunction and the efficacy of treatment with water-soluble compounds in cerebellar neurodegenerative disease.

We present a biochemical and behavioral protocol to evaluate the efficacy of mitochondria-targeted water-soluble compounds for the treatment of Spinocerebellar ataxia type 1 (SCA1) and other cerebellar neurodegenerative diseases.

治疗SCA1小鼠水溶性化合物，以非特异性升压线粒体功能

Mitochondrial dysfunction plays a significant role in the aging process and in neurodegenerative diseases including several hereditary spinocerebellar ...

一个简单的综合评分系统评估小脑共济失调的小鼠模型的表型

摘要

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此视频中的章节

临床试验和脊髓去除在肌萎缩性侧索硬化症的小鼠模型（ALS）

神经肌肉疾病小鼠模型行为花型的低成本盖特分析

一种研究甲状腺激素对小脑功能影响的通用行为方法

一种研究甲状腺激素对小脑功能影响的通用行为方法

一种研究甲状腺激素对小脑功能影响的通用行为方法

安格曼综合征小鼠模型的行为表征

安格曼综合征小鼠模型的行为表征

安格曼综合征小鼠模型的行为表征

一个快速的表型神经评分系统在ALS的SOD1-G93A小鼠模型评估疾病进展

治疗SCA1小鼠水溶性化合物，以非特异性升压线粒体功能