We would like to acknowledge Beth Levine for critically reviewing the manuscript, Valerie Tassinari for developing the genotyping protocol, and Matt Ferola and Carlos Maya for their exceptional animal care.

所有实验都按照由卫生指南的国家机构的护理和使用动物的所述的协议进行，并批准了ALS TDI的机构动物护理和使用委员会（IACUC）。 1，动物，房屋和研究设计注:在SOD1-G93A小鼠菌落从高拷贝B6SJLTgN（SOD1G93A）1Gur应变最初由格尼等7产生的派生。菌落目前正在通过杂交的转基因C57BL / 6-SJL雄性野生型C57BL / 6-SJL F1雌性保持。通过杂交SJL雄性与雌性B6产生的F1动物。小鼠被运送到ALS TDI每周在35-45日龄。 <ol><li>允许小鼠至少一周分配给一个研究之前来适应环境的动物设施（12小时亮/暗循环，在18-23℃，40-60％湿度的温度下）。 </li><li>宅男SOD1-G93A麦克风每笼ê一个鼠标，因为群养的男性往往会打。宅女SOD1-G93A小鼠长达两个每笼。提供丰富环境，塑料小屋的形式。提供食物和水随意，除非小鼠是研究需要药物施用在饮用水/食品的一部分。在这种情况下，理性，记录食物和水。 </li><li>开始围绕日龄50药物疗效研究，其中包括64产仔匹配的性别平衡的小鼠（每个队列32）。最后，确保在进行研究的研究人员正在失明的实验条件。 注意:建议，一个典型的药物功效研究开始于年龄天50但是根据治疗介入的可能是优选的启动研究较早或较晚的类型。 注:研究用32只小鼠的队列通电检测性别特异性药物效果。如果没有性别区分的比较是必需的，每个队列24只应该足以检测药物的整体效果。</li></ol> 2.体重<ol><li>记录体重为所有小鼠在研究中每天用一克的十分之一的精度。注:体重是任何不适，可能会导致慢性药物治疗的疾病进展和的敏感指标。 </li><li>小鼠体重变化相当一天的过程中，作为在早晨更高，从而每天大约在同一时间记录下来。 </li></ol> 3.神经评分系统（NeuroScore） <ol><li>评估神经学得分（NS）的每只小鼠每日通过以下三种情况下观察鼠标顺序执行:a）该小鼠是由它的尾巴悬挂，b）该小鼠被允许行走，轻瘫C的发病后）的鼠标放置在其一侧。 <ol><li>对于悬尾实验，按住鼠标大约1.5"从尾巴在他们的家笼的线顶部的基地1-2秒，远离菜盆，而OBS欧文的后肢。重复测试悬浮液3次，并记录最一致的结果。 </li><li>对于行走测试，将鼠标放在一个干净的表面提供了一些牵引力和步行距离25厘米（例如，纸巾贴，以防止打滑）。让鼠标一共75厘米（纸巾的3倍长）边走边观察其步态。 </li><li>对于"翻正反射"试验，将鼠标在其左侧或右侧，并用秒表测量才能纠正自己独立站起双方的时间。注:本次测试中仅有一个实验是必需的。 </li></ol></li><li>确定每个后肢的NS（左或右）独立地在一个范围从0到4。注意:例如评分为0，1表示左后肢NS为0，而右后肢NS为1。 </li><li>分配对应的观测（摘要表 1中提供）下面的一组神经学得分（0-4）:<ol><li>分配NS 0（预症状）如果符合下列条件:当鼠标的尾巴悬吊时，后肢呈现正常喷射也就是说，它完全伸展远离横向中线和它保持在这个位置上，持续2秒或更长的时间。当鼠标被允许行走，正常步态是观察。 </li><li>分配NS 1（首发症状）是否遵守以下规定:当鼠标的尾巴悬挂，后肢呈现异常扇， 也就是说，它被折叠或部分向外侧中线倒塌或在悬尾它颤抖或是缩回/抱住。当鼠标被允许行走，正常或稍缓慢步态是观察。 </li><li>分配NS 2（麻痹性痴呆的发病），如果符合下列条件:当鼠标的尾巴悬挂，后肢部分或完全倒塌，很多不延长。 （有可能仍然是关节运动）。当鼠标被允许行走，后肢用于向前MOT然而离子脚趾在一张90厘米的步行向下卷曲至少两次或任何部分的脚的沿笼底/表拖动。当鼠标被放置在它的左侧和右侧，它能够从两侧在10秒内权利本身。 </li><li>分配的NS 3（麻痹）如果符合下列条件:当鼠标的尾巴悬吊，有刚性的麻痹的后肢或最小关节运动。当鼠标被允许行走有向前然而后肢不被用于向前运动的运动。当鼠标被放置在它的左侧和右侧，它能够从两侧在10秒内权利本身。注:很少，瘫发病后，尿液中的水分可能会出现在后肢。如果不进行治疗尿水分，可能会导致尿"烧钱"和皮肤损伤。治疗尿水分被裁剪的头发，应用温水浸泡去除尿液，并轻轻擦干。如果皮肤损伤是存在适用antibio抽动软膏。 </li><li>指定NS 4（人文终点），如果符合下列条件:当鼠标的尾巴悬挂，有刚性瘫痪在后肢。当鼠标被允许行走，没有向前运动。当鼠标被放置在其左，右手边是不是能够在来自任何一方的10秒权利本身。 也就是说，没有翻正反射。 </li></ol></li></ol> 4.数据录入和分析<ol><li>期间药效研究的过程中，进入收集每只小鼠成实验室信息管理系统（LIMS）或到市售的表计算软件的数据。 注:ALS TDI开发的LIMS软件是从内部研究收集的所有数据的中央存储库，使这些数据随着时间的推移ALS TDI研究者方便地访问。 LIMS跟踪任何个人的鼠标，凭借其独特的ID，回到它的血统，体重或神经功能评分在所有研究日，包括在其一生中指出的其他任何意见。 LIMS最终产生其用于后续分析的电子表格。 </li><li>当完成一个药效研究， 即，当所有的小鼠到达人道终点，使用电子表格软件（见表材料）通过平均在左右后肢的NS来计算合并的NS每只小鼠，每天的。接着，计算平均年龄在每个NS对每个小鼠每只小鼠花费在特定的NS的天数。 </li><li>通过从电子表格软件导入数据的统计软件分析在一段时间内的NS总体变化（见表材料）。符合这些数据的使用顺序逻辑回归的模型估计预期的NS，中位年龄在NS和药物治疗作为回归的影响。使用对数秩卡方检验估计的影响似然概率，并确定在NS进展的药物治疗效果是否是统计LY显著（见一步一步的指示补充编码文件）。 注:p值&lt;0.05被认为是统计显著。 注意:统计分析的该方法尊重的NS数据并向NS轨迹单个小鼠的可变性质的序号性质。这是首选，因为个人NS轨迹可以明显不同导致NS时关联组平均数是不充分的反映NS响应。此外，在某些情况下，NS会波动，特别是当在零到一的范围内。 </li><li>最后，使用图形软件（见表材料）生成"序预计NS VS平均年龄NS"图表药物治疗组和对照组动物（ 图2，见补充编码文件一步一步的说明）。使用这些图形由y轴插补疾病发作（ 我的年龄中位数为方便评估药物治疗组和对照组之间的变化 。即，NS 2）上x轴。 注意:在两组中位年龄的差异代表药物引起的移位和被当作效果幅度的实际措施。 注:在预期NS与年龄中位数曲线表明减缓症状的疾病进展右移。相反，在预期的NS对位年龄曲线向左偏移指示加速症状的疾病的进展。 </li></ol>

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The authors have nothing to disclose.

在这份报告中，我们描述了一个快速和简单的视频协议，如果应用得当，能够可靠地评估SOD1-G93A小鼠的疾病进展和确定药物引起的变化。尽管不同群体开发的表型的评分系统对SOD1-G93A小鼠8-10，他们往往不提供关于该过程足够的细节和不足进行复制。其结果，评分系统很少使用，开发他们的特定的组之外。在这份报告中，用视频详细手稿组合最大程度地减少评分含糊和跨实验者变异从而允许该协议由其他实验室采用，使研究发生在不同的设置之间的比较。 本报告中称为NeuroScore中所描述的神经评分系统，其中产生的精确值归因于体征和sy的集有序数据mptoms具有超出建立位列疾病的严重程度没有意义。在这种情况下，NS 0是最不严重的疾病状态和表示一个鼠标与正常后肢功能而NS 4是小鼠中最严重的疾病状态或接近发病继发于神经肌肉疾病。尽管评分系统使得采用对症阈值，每个序号NS值涵盖了广泛的其中许多被捕获的视频和本报告中的可能临床表现谱。 NS 1包括的疾病在SOD1-G93A小鼠中的早期症状，其可以有很大的不同，从小鼠到小鼠的频谱。一些小鼠表现出与倒塌朝中线，而另一些则后肢震颤，有的甚至表现出后肢回缩或扣紧后肢异常张开。此外，在疾病的早期阶段的一些小鼠的NS可能波动为NS 1和0之间的若干天无一例外虽然，在SOD1-G93A小鼠中的频率的NS 1随时间增加，它包括多描述的不同表型的多。平均来说，雄性和雌性的SOD1-G93A小鼠中通常花费37分别天和28天在这个阶段。 NS 2表示轻瘫（轻微或局部麻痹）的发作和提供不可否认的证据，病理的存在。虽然这是很平常的野生型小鼠有时表现为NS 1 NS 2只发生在SOD1-G93A小鼠。类似于NS 1，NS 2捕获的症状和体征的频谱。在早期阶段呈现NS 2的鼠标动态，精力充沛。 NS 2更先进的演示表明与移动一个非常有限的能力的老鼠。在后肢轻瘫先进可以防止老鼠从设置在固定框架顶部到达食物和水，所以食物丸应放置在笼的底部或应提供水合食品。另外，较长的吸管应放在水瓶。 SOD1-G93A小鼠typicaLLY花费5天这个阶段。 NS 3表示完整的后肢瘫痪，并通过所有帐户是最简单的表型鉴定和得分。由NS 3包围的麻痹症状可以是松弛或痉挛性，但通常是痉挛。由这一点上，小鼠是非常虚弱，要经常进行监测。平均来说，SOD1-G93A小鼠花费1至2天在这个阶段。 NS 4表示没有"翻正反射"不完全依赖于像NS 0-3后肢的功能，而是它体现了鼠标的整个身体健康。小鼠表现出正常的"翻正反射"的能力，需要的动力，能源，而且多个肌群包括后肢，前肢和腹部的功能。要注意在这些小鼠中疾病进展经常是不对称的和鼠标时放置在一侧米，可以成功地完成正位反射是很重要的AY是不能这样做的时候放在对方。因此重要的是，对于动物的人道待遇和研究的完整性，对双方进行测试，如果它没有权利本身两侧安乐死鼠标。 NS 4定义安乐死之日起，默认情况下与生存数据直接对齐。但是，总的疾病进展，通过生理盐水有序回归分析所定义，可能不完全重叠与Kaplan-Meier生存曲线和体重的数据。某些治疗干预可能影响疾病发作或症状，但不影响整体的寿命。 而没有深入本报告中所讨论的，体重可以是可被组合使用以NS评估疾病发作和进展中的SOD1-G93A但它可以通过在饮食差异偏斜的另一项措施，并通过疗法引起水肿或影响脂肪代谢11,12。 13，而不是后肢运动障碍，不是由NeuroScore拍摄的表型。此协议的另一个限制是，它是专为，只适用于高仿SOD1-G93A小鼠。根据我们的经验，在ALS的其他小鼠模型的临床表现的疾病，如低拷贝SOD1-G93A小鼠14或 TDP-43小鼠15,16，比在高拷贝的SOD1-G93A小鼠和这个NS系统不同可能并不适合那些模型。因此，现有来确定小鼠的转基因拷贝数，以将其分配到研究和验证它等于高拷贝阳性对照小鼠的是重要的。我们不管，带来的好处荷兰国际集团这个神经评分系统超过了它的局限性，这个视频协议可以作为一个很好的训练工具，用于ALS研究人员。

自从它在90年代中期开发的SOD1-G93A转基因小鼠模型已经肌萎缩侧索硬化症（ALS）1的最广泛使用的动物模型。这种转基因小鼠模型被基因工程化以过表达人铜/锌超氧化物歧化酶1（SOD1）基因携带有ALS相关甘氨酸至丙氨酸的突变在氨基酸93（G93A）的突变体形式。的G93A突变是许多突 ​​变的SOD1基因，它们共同化妆大约五分之一的家族性ALS例2之一。 在SOD1-G93A模式已经成为了ALS药物开发研究的一个主力，因为除了要ALS其明显的遗传链接，这概括了许多ALS在人体内的病理特点，如运动神经元丧失，肌肉逐渐萎缩无力，与最终麻痹而死亡3。虽然，这是常有用的转基因动物模型的情况下，有一种内在的双向与SOD1-G93A小鼠相关ological变化。 Scott等。已经确定，需要在药物功效的研究设计，以克服由生物变异4中创建的噪声至少24垫料匹配性别平衡小鼠的队列。大量在这些研究中与侵袭性疾病进展和需要日常监测组合需要的动物的禁止使用的精心设计的，耗时的和潜在的压力的技术，用于测量神经肌肉强度和功能（例如，肌电图，握力，旋转棒等）。相反，它强调需要一个体内筛选工具，可靠地评估了一个快速的方式SOD1-G93A小鼠的神经肌肉功能。 在ALS TDI的协议被开发，允许SOD1-G93A小鼠的神经肌肉功能的每只小鼠不到30秒，平均可靠的评估。该协议encompa小规模企业后肢功能的使用是该报告在媒体和视频描述了一个简单的神经评分系统（NeuroScore）每天评估。 NeuroScore专注于后肢功能，因为后肢赤字是最早报道的疾病在SOD1-G93A小鼠5,6神经的迹象。此外，通过ALS TDI开发的协议提供了发病轻瘫（轻微或局部麻痹），进展和麻痹的严重程度的公正评估，这是不够敏感，以确定在疾病进展的药物引起的变化。

<table><tbody><tr><td>B6SJLTgN(SOD1G93A)1Gur strain</td><td>The Jackson Laboratory, Bar Harbor, ME</td><td>002726</td><td>Strain of mice used in this study</td></tr><tr><td>Breeders</td><td>Biomedical Research Models, Inc., Worcester, MA</td><td></td><td>Colony maintainance</td></tr><tr><td>Scale</td><td>Navigator OHAUS, Parsippany, NJ</td><td>Model N14120</td><td>Used to weigh mice</td></tr><tr><td>Nutra-GelH</td><td>Bio-Serv, Flemington, NJ</td><td>#S4798</td><td>Wet food provided to sick mice</td></tr><tr><td>Irradiated Lab Animal Diet</td><td>Harlan, Indianapolis, IN</td><td>2918-111914M</td><td>Chow diet</td></tr><tr><td>Lab-grade Sani-chips</td><td>Harlan Teklad, Indianapolis, IN</td><td>7090</td><td>Bedding</td></tr><tr><td>JMPH&amp;nbsp;</td><td>SAS Institute, Inc., SAS Campus Drive, Cary, NC</td><td>&amp;nbsp;v10.0.2</td><td>Statistical software</td></tr><tr><td>Microsoft Excel</td><td>Microsoft, One Microsoft Way, Redmond, WA</td><td>Excel 2013 (v 15.0)</td><td>Spreadsheet software</td></tr><tr><td>GraphPad Prism 5</td><td>GraphPad software Inc., La Jolla, CA</td><td>v5.4</td><td>Graphing software</td></tr><tr><td>Mouse Huts</td><td>Bio-Serv, Flemington, NJ</td><td>K3272</td><td>For environmental enrichment</td></tr></tbody></table>

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.

神经功能评分数据为90男，94女未治疗的SOD1-G93A小鼠就读于ALS TDI在2014年进行了评价。结果表明，雄性小鼠通常具有比雌性小鼠更具侵略性的疾病进展，如由在NS 1的比例更大其寿命相比，女性。 NS 2和NS 3发生跨性别大致相等的频率。 NS 0被从分析中排除，因为它被认为是一个"正常"表型。 NS 4也被排除在外，因为根据定义，它只能出现一次在SOD1-G93A小鼠的生存期（ 表2和图1）。 NS数据从ALS疾病进展药效药理试验化合物TDI173在SOD1-G93A小鼠研究中提出（每组n =性别/治疗16; 64总只小鼠）。在图2中，神经系统疾病进展的速率是作为捕获数据c的顺序回归分析aptured使用本文详述的NeuroScore协议。 TDI173处理的小鼠进行比较，以载体处理的对照。无统​​计学观察显著的差异时，分别进行了分析，但男女两性的时候被合并统计显着性达到。生存的Kaplan-Meier分析，尽管示出了类似的趋势作为有序Logistic统计分析，是不够敏感以揭示显著差异甚至当性别被合并（ 图3）。 <table border="0" cellpadding="0" cellspacing="0"><tbody><tr><td> NS 1 </td><td>后肢呈现异常扇， 也就是说，它是倒塌或部分倒塌朝外侧中线或在悬尾它颤抖或是缩回/紧握。 </td><td>正常或略慢的步态。 </td><td> NA </td></tr><tr><td> NS 2 </td><td>后肢部分或完全倒塌，很多不延长。可能有ST生病是关节运动。 </td><td>后肢用于向前运动然而脚尖期间75厘米步行或任何部分的脚向下卷曲至少两次沿笼底/表拖动。 </td><td>鼠标是能够从双方在10秒内权利本身。 </td></tr><tr><td> NS 3 </td><td>刚性瘫痪后肢或最小关节运动。 </td><td>向前运动然而后肢不被用于前向运动。 </td><td>鼠标是能够从双方在10秒内权利本身。 </td></tr><tr><td> NS 4 </td><td>刚瘫痪的后肢。 </td><td>不向前移动。 </td><td>鼠标不能够在来自任何一方的10秒权利本身。 </td></tr></tbody></table> 表1:用于分配的神经评分表型的观察总结。 <table border="0" cellpadding="0" cellspacing="0"><tbody><tr><td></tD&gt; <td colspan="2"> NS1频率</td><td colspan="2"> NS2频率</td><td colspan="2"> NS3频率</td></tr><tr><td></td><td>男</td><td>女</td><td>男</td><td>女</td><td>男</td><td>女</td></tr><tr><td>小鼠数</td><td> 90 </td><td> 94 </td><td> 90 </td><td> 94 </td><td> 90 </td><td> 94 </td></tr><tr><td>起码</td><td> 12 </td><td>五</td><td> 1 </td><td> 1 </td><td> 0 </td><td> 0 </td></tr><tr><td> 25％ </td><td> 29 </td><td> 16 </td><td> 4 </td><td> 4 </td><td> 0 </td><td> 0 </td></tr><tr><td>中位数</td><td> 37 </td><td> 27.5 </td><td>五</td><td>五</td><td> 1 </td><td> 1 </td></tr><tr><td> 75％ </td><td> 44 </td><td> 37.5 </td><td> 7.5 </td><td> 6.5 </td><td> 2.5 </td><td> 3 </td></tr><tr><td>极大</td><td> 61</td><td> 60 </td><td> 28 </td><td> 11 </td><td> 8 </td><td> 9 </td></tr></tbody></table> 表2:NS数据每天日龄50拿下，直至死亡未治疗的SOD1-G93A小鼠的频率分析。分别计算NS 1，2和3的分布。 <img alt="图1" src="/files/ftp_upload/53257/53257fig1.jpg" /> 图1。天每只小鼠来自未处理的SOD1-G93A小鼠中的细胞集落的群中的每一异常NeuroScore花费的数目的图形表示过程2014（雄性每组90，女94例）的得分频率为同小鼠被表示为每一种可能的NS。 NS 4省略，因为只能事件发生在每只小鼠的生存期的定义一次。 <a href="https://www.jove.com/files/ftp_upload/53257/53257fig1large.jpg" target="_blank">请点击此处查看大versio</a><html> n这个数字。 <img alt="图1" src="/files/ftp_upload/53257/53257fig2.jpg" /> 图2典型的有序Logistic统计分析比较TDI173治疗的小鼠（红色线），以垃圾匹配的车辆处理的对照小鼠的神经系统疾病的发展曲线。（蓝线;报导的P值代表数秩卡方检验） <a href="https://www.jove.com/files/ftp_upload/53257/53257fig2large.jpg" target="_blank">请点击这里查看该图的放大版本。</a> <img alt="图1" src="/files/ftp_upload/53257/53257fig3.jpg" /> 图3比较TDI173治疗的小鼠（红色线），以垃圾匹配的车辆处理的对照小鼠的寿命典型Kaplan-Meier分析图（蓝线;报导的P值代表数秩检验）。jove.com/files/ftp_upload/53257/53257fig3large.jpg"目标="_空白"&gt;点击此处查看该图的放大版本。

Watch this Scientific Journal Video about A Quick Phenotypic Neurological Scoring System for Evaluating Disease Progression in the SOD1-G93A Mouse Model of ALS at JoVE.com

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

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 ...

a-quick-phenotypic-neurological-scoring-system-for-evaluating-disease

Nanyang Technological University

Research

JoVE Journal

Medicine

19.4K 次观看.  ALS Therapy Development Institute. This video protocol describes a sensitive, reliable, and quick method for evaluating the neuromuscular deficits in a transgenic mouse model of amyotrophic lateral sclerosis.

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

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

We describe a protocol for the rapid and sensitive quantification of disease severity in mouse models of cerebella ataxia. It is derived from previously published phenotype assessments in several disease models, including spinocerebellar ataxias, Huntington s disease and spinobulbar muscular atrophy. Measures include hind limb clasping, ledge test, gait and kyphosis. Each measure is recorded on a scale of 0-3, with a combined total of 0-12 for all four measures. The results effectively discriminate between affected and non-affected individuals, while also quantifying the temporal progression of neurodegenerative disease phenotypes. Measures may be analyzed individually or combined into a composite phenotype score for greater statistical power. The ideal combination of the four described measures will depend upon the disorder in question. We present an example of the protocol used to assess disease severity in a transgenic mouse model of spinocerebellar ataxia type 7 (SCA7).
Albert R. La Spada and Gwenn A. Garden contributed to this manuscript equally.

We describe a protocol for the rapid and sensitive quantification of disease severity in mouse models of cerebellar ataxia. Measures include hind limb clasping, ledge test, gait and kyphosis. This protocol effectively discriminates between affected and non-affected individuals, and detects the progression of affected individuals over time.

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 ...

科研

神经科学

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 ...

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

Measuring Progressive Neurological Disability in a Mouse Model of Multiple Sclerosis

After intracerebral infection with the Theiler's Murine Encephalomyelitis Virus (TMEV), susceptible SJL mice develop a chronic-progressive demyelinating disease, with clinical features similar to the progressive forms of multiple sclerosis (MS). The mice show progressive disability with loss of motor and sensory functions, which can be assessed with multiple apparatuses and protocols. Among them, the Rotarod performance test is a very common behavioral test, its advantage being that it provides objective measurements, but it is often used assuming that it is straightforward and simple. In contrast to visual scoring systems used in some models of MS, which are highly subjective, the Rotarod test generates an objective, measurable, continuous variable (i.e., length of time), allowing almost perfect inter-rater concordances. However, inter-laboratory reliability is only achieved if the various testing parameters are replicated. In this manuscript, recommendations of specific testing parameters, such as size, speed, and acceleration of the rod; amount of training given to the animals; and data processing, are presented for the Rotarod test.

An optimized testing protocol is presented in this paper for the Rotarod performance test, used for measuring progressive neurological disability in TMEV-infected mice.

在多发性硬化症的小鼠模型测量进行性神经功能残疾

After intracerebral infection with the Theiler's Murine Encephalomyelitis Virus (TMEV), susceptible SJL mice develop a chronic-progressive demyelinating ...

Assessment of Spontaneous Alternation, Novel Object Recognition and Limb Clasping in Transgenic Mouse Models of Amyloid-&#946; and Tau Neuropathology

Here we describe a staged, behavioral testing approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional motor behaviors in transgenic mouse models of &#946;-amyloidosis and tauopathy. The paradigm includes tests for spontaneous alternation in a Y-maze, novel object recognition, and limb clasping. These tests were selected because they: 1) interrogate function of cognitive or motor domains and the correlate neural circuitry relevant to the human disease state, 2) have clearly defined endpoints, 3) have easily implementable quality control checks, 4) can be run in a moderate throughput format, and 5) require little intervention by the investigator. These methods are designed for investigators looking to screen compounds for activity in short-term and working memory tasks, or functional motor behaviors associated with Alzheimer's disease mouse models. The methods described here use behavioral tests that engage a number of different brain regions including hippocampus and various cortical areas. Investigators that desire cognitive tests that specifically assess cognition mediated by a single brain region could use these techniques to supplement other behavioral tests.

Described here is a staged, behavioral screening approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional motor behaviors in transgenic mouse models of &#946;-amyloidosis and tauopathy. These methods are optimized to screen compounds for activity in short-term and working memory tasks.

自发交替评估，淀粉样蛋白β和脑神经病理学转基因小鼠模型中的新物体识别和肢体紧扣

Here we describe a staged, behavioral testing approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional ...

Repeated Measurement of Respiratory Muscle Activity and Ventilation in Mouse Models of Neuromuscular Disease

Accessory respiratory muscles help to maintain ventilation when diaphragm function is impaired. The following protocol describes a method for repeated measurements over weeks or months of accessory respiratory muscle activity while simultaneously measuring ventilation in a non-anesthetized, freely behaving mouse. The technique includes the surgical implantation of a radio transmitter and the insertion of electrode leads into the scalene and trapezius muscles to measure the electromyogram activity of these inspiratory muscles. Ventilation is measured by whole-body plethysmography, and animal movement is assessed by video and is synchronized with electromyogram activity. Measurements of muscle activity and ventilation in a mouse model of amyotrophic lateral sclerosis are presented to show how this tool can be used to investigate how respiratory muscle activity changes over time and to assess the impact of muscle activity on ventilation. The described methods can easily be adapted to measure the activity of other muscles or to assess accessory respiratory muscle activity in additional mouse models of disease or injury.

This paper introduces a method for repeated measurements of ventilation and respiratory muscle activity in a freely behaving amyotrophic lateral sclerosis (ALS) mouse model throughout disease progression with whole-body plethysmography and electromyography via an implanted telemetry device.

在神经肌肉疾病的模型小鼠呼吸肌肉活动和通风的重复测量

Accessory respiratory muscles help to maintain ventilation when diaphragm function is impaired. The following protocol describes a method for repeated ...

In Vivo Electrophysiological Measurement of Compound Muscle Action Potential from the Forelimbs in Mouse Models of Motor Neuron Degeneration

Assessing the functionality of the nerve axon provides detailed information on the progression of neuromuscular disorders. Electrophysiological recordings provide a sensitive approach to measure nerve conduction in humans and rodent models. To broaden the technical possibilities for electromyography in mice, the measurement of compound muscle action potentials (CMAPs) from the brachial plexus nerve in the forelimb using needle electrodes is described here. CMAP recordings after stimulating the sciatic nerve in hindlimbs have been previously described. The newly introduced method here allows for the evaluation of the nerve conductivity at an additional site, and thus provides a more profound overview of the neuromuscular functionality. The technique provides information on both the relative number of functional axons and the myelination level. Thereby, this method can be applied to assess both axonal diseases as well as demyelinating conditions. This minimally invasive method does not require extraction of the nerve and therefore it is suitable for repeated measurements for longitudinal follow-up in the same animal. Similar recordings are performed in clinical setups to emphasize the translational relevance of the method.

In Vivo Electrophysiological Measurement of Compound Muscle Action Potential from the Forelimbs in Mouse Models of Motor Neuron Degeneration

The measurement of nerve conduction is a useful tool to assess mouse models of neurodegeneration but it is frequently only applied to stimulate the sciatic nerve in hindlimbs. Here, we describe a technique to measure compound muscle action potential (CMAP) in vivo in the mouse forelimb muscles innervated by the brachial plexus.

体内运动神经元变性小鼠前肢肌作用电位的电生理测量

Assessing the functionality of the nerve axon provides detailed information on the progression of neuromuscular disorders. Electrophysiological recordings ...

脊髓组织学染色用于评估 EAE 疾病严重程度

Scoring Central Nervous System Inflammation, Demyelination, and Axon Injury in Experimental Autoimmune Encephalomyelitis

Experimental autoimmune encephalomyelitis (EAE) is a common immune-based model of multiple sclerosis (MS). This disease can be induced in rodents by active immunization with protein components of the myelin sheath and Complete Freund&#39;s adjuvant (CFA) or by the transfer of myelin-specific T effector cells from rodents primed with myelin protein/CFA into na&#239;ve rodents. The severity of EAE is typically scored on a 5-point clinical scale that measures the degree of ascending paralysis, but this scale is not optimal for assessing the extent of recovery from EAE. For example, clinical scores remain high in some EAE models (e.g., myelin oligodendrocyte glycoprotein [MOG] peptide-induced model of EAE) despite the resolution of inflammation. Thus, it is important to complement clinical scoring with histological scoring of EAE, which also provides a means to study the underlying mechanisms of cellular injury in the central nervous system (CNS).
Here, a simple protocol is presented to prepare and stain spinal cord and brain sections from mice and to score inflammation, demyelination, and axonal injury in the spinal cord. The method for scoring leukocyte infiltration in the spinal cord can also be applied to score brain inflammation in EAE. A protocol for measuring soluble neurofilament light (sNF-L) in the serum of mice using a Small Molecule Assay (SIMOA) assay is also described, which provides&#160;feedback on the extent of overall CNS injury in live mice.

作者聚焦：揭示多发性硬化症中肥胖与自身免疫性炎症之间的通路 

Experimental autoimmune encephalomyelitis (EAE) serves as an animal model of multiple sclerosis. This article describes an approach for scoring spinal cord inflammation, demyelination, and axonal injury in EAE. Additionally, a method to quantify soluble neurofilament light levels in the mice serum is presented, facilitating the assessment of axonal injury in live mice.

对实验性自身免疫性脑脊髓炎中的中枢神经系统炎症、脱髓鞘和轴突损伤进行评分

Experimental autoimmune encephalomyelitis (EAE) is a common immune-based model of multiple sclerosis (MS). This disease can be induced in rodents by ...

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

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此视频中的章节

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

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

在多发性硬化症的小鼠模型测量进行性神经功能残疾

自发交替评估，淀粉样蛋白β和脑神经病理学转基因小鼠模型中的新物体识别和肢体紧扣

在神经肌肉疾病的模型小鼠呼吸肌肉活动和通风的重复测量

体内运动神经元变性小鼠前肢肌作用电位的电生理测量

对实验性自身免疫性脑脊髓炎中的中枢神经系统炎症、脱髓鞘和轴突损伤进行评分

对实验性自身免疫性脑脊髓炎中的中枢神经系统炎症、脱髓鞘和轴突损伤进行评分

对实验性自身免疫性脑脊髓炎中的中枢神经系统炎症、脱髓鞘和轴突损伤进行评分