威康信托基金会提供开放存取资金牛津大学。罗伯特·迪肯资助，由威康信托基金资助WT084655MA牛津OXION组的成员。

1。设备对于所有的测试，软垫的表面放置在该基地的设备，以减轻任何脱落的小鼠。注意这是未示出在附图中，为了不遮挡任何设备的一部分。数百小鼠在试验过程中，正常和突变的，从来没有观察到任何损伤所造成的下降，从这样的装置，它通常是30-50厘米高。此外，请务必清洗和消毒的设备之间测试每个鼠标。 1.1 Rotarod 该设备在市场上的商业版本有几个，但有些缺点，如没有足够的速度加快，检测运动不协调（而不是耐力）。另一种常见的故障是杆以上的基地不够高，导致老鼠趋于跌落或跳下。 图1中所示的小鼠旋转的设计的作者，牛津大学药理学部门的工程部分。杆的直径3厘米，30厘米以上的装置的基础上。在一系列沿纵向轴线平行的脊的表面滚花，使老鼠所困扰，它有效地（ 图2）。 NB脊的深度是一个重要的细节，如果鼠标可以不得到良好的抓地力测试将是非常多困难;另一方面，如果不太好握鼠标将杆周围的“车轮”，被动地持有到它。对于“业余rotarod决策者”，它可能是有用的，知道那滚花木质dowelling的小鼠的适当大小是市面上，这当然需要治疗，具有良好的防水漆使用前。的起始速度被调整到每分钟4转，20转/分的加速率。最大速度为每分钟40转。 两个法兰防止鼠标离开杆。它们是直径30厘米（这也许可以减少到20厘米）。他们的分离被设定为6厘米（最大），但可能需要调整子成年小鼠小，如果他们往往在杆转身。 1.2三重单杠（图3） 该酒吧的材质有黄铜，38厘米长，板凳上表面49厘米以上，在每年年底的木制支撑柱。列固定到一个沉重的木基地。三杆直径：2，4和6毫米。 2毫米的酒吧是我们使用的标准之一，但许多小鼠达到最高分数。因此，大直径的棒材已计入在细化测试老鼠一样，不能握这些企图。 1.3静态棒五木销钉或杆粗细不一（35（1杆），28，22，15和9（5杆）毫米直径）各60厘米长的固定由G-钳实验室架子棒等水平突出成的空间（ 图4）。年底标志杆附近的长凳上有一个10厘米到底，表示终点线。在地板上方的杆的高度为60厘米。 如果需要一个较短和较详细的测试，只能使用其最大，最小和中间棒。凡Dellen 等14只用一个单一的杆亨廷顿氏病在小鼠中的进展进行评估。 1.4双杠两个平行的钢条1米长，直径4毫米，被固定在其端部（ 图5）的木制的支承柱30毫米（在其中心）。酒吧是60厘米以上的地板。 2。程序对于所有的测试，把老鼠的试验室测试前5-20分钟，以确保它们是完全清醒的。作为一般规则，以允许恢复肌肉力量和回归到正常水平的觉醒，老鼠休息后返回家笼每个电机测试。/ P&gt; 2.1 Rotarod 与一开始设定的小鼠旋转速度为每分钟4转，加速速率每分钟20转/分钟。按住鼠标的尾巴，把它放在旋转杆，背对的旋转方向，所以它具有向前走，保持直立。这是最简单的做，如果鼠标被带起朝向杆在水平45°以下的角度，试图降低来自上述鼠标将导致扩散其后腿和把握的凸缘的边缘。快速释放鼠标时，它几乎是接触杆，只是向前和棒上方的上死点中心，从而使其能够很容易地把持棒。 （可能更容易和更可靠）技术是使用薄木钉（10-15毫米）。按住鼠标，在一方面，降低它的定位销上，平行于长轴。木钉成鼠标的头部向下约30°，给它更多的时间来夹住小鼠旋转，一旦它被释放。放下鼠标，日之间的木钉ê法兰的小鼠在Rotarod，一次在旋转杆拉的木钉向下远离鼠标，这将掌握杆，往前走。 在10秒后，将鼠标杆，开始加速（只要鼠标正面临着：如果没有，等到它确实面临着加速开始前），并注意鼠标的速度脱落。如果脱落，前10秒，请注意时间秋天，再试一次，最多三次总记录的速度10秒点后的首次下降。然而，落在前5秒，是由于被实验者配售差不应该被记录。的平均下落速度为基准，而不是使用的最大速度，这额外的练习校正鼠标接收过程中发生故障的运行，其中假设将有助于性能。因此，鼠标落在前10秒一次，然后保持至12转第二次得分（4 +12）/ 2 = 8转。鼠标两次，然后下降停留在10转分数（4 +4 +10）/ 3 =每分钟6转。如果鼠标没有握在10秒三次分配一个得分4转。 其中一个问题是，老鼠有时会停止向前走，而不是紧紧握住杆，因此，他们被动地翻腾轮，虽然大多数最终脱落。因此，它可能是有用的，也注意到第一个这样的反转发生时的速度。一种可能的解决办法可能是使用直径较大或不太明显的脊杆。 2.2单杠老鼠发现最容易掌握的窄2毫米bar，最好是先测试他们在这一个。握住鼠标的尾巴，将其放置在板凳上设备前，迅速向后滑动约20厘米（垂直对齐到酒吧），迅速提高并让它抓住单杠，在中央点它的前爪，然后松开尾巴，同时启动stopclock。这个C难以有效实现;一些老鼠抓地力更好，如果尾部突然被释放，如果他们觉得自己仍然支持，它们可能会握。标准点是一个秋天从酒吧，鼠标达到前年底列的酒吧之一，或时间，直到一个前爪触及一列。最大测试时间（截止时间）为30秒。 如果将鼠标失败来把握吧妥善第一次和这似乎是由于实验者的技术，而不是鼠标，尝试再次（后稍作休息，同时测试了另外一个或两个小鼠）和不记录在今年秋天。如果鼠标落在前5秒，这显然是不由于配售差由实验者，重复三次，试图得到一个&gt; 5秒得分。如果&gt; 5秒在第二次尝试，不要做第三个审判。采取的最好成绩为基准。鼠标反复无法支持&gt; 5秒的成绩只有1。 如果三杆正在使用的版本，如果鼠标在此第2毫米bar成绩5就可以进行测试上较厚的酒吧，经过短暂的休息时间，而另外一个或两个小鼠进行测试。为2毫米bar为4和6毫米酒吧的评分系统是相同的，最终得分的累积总数。因此，一个鼠标，5分2毫米的酒吧，但下降后，从4毫米的酒吧13秒的成绩5 +3 = 8。 2.2.1 单杠得分：前两个间隔低于过去两年，因为一旦老鼠已经初步掌握了任务，他们不太可能回落： 之间1-5秒= 1 下降6-10秒= 2 在11-20秒之间下降= 3 在21-30秒之间下降= 4 后30秒= 5 酒吧的支持，而不配售前爪下降= 5 NB选用4和6毫米的酒吧是一个新的除了我们的协议中，我们还没有广泛与他们的经验。 2.3静态棒将鼠标最广泛杆（鼻尖一个头的长度到底是理想的）的尽头。采取两项措施：定向时间（时间采取从起始位置朝货架方向180°）和过境时间（前往货架端（鼻超出10厘米从架子上的标志杆底）所花费的时间。 取向依赖于鼠标保持直立;如果事实颠倒低于杆紧贴，任意分配（为统计目的）的分数为120秒的最大取向。不要测试小棒，在途时间也变成最大值作为过渡直立鼠标显然是一个过渡，同时挂倒挂更好地协调。 取向后，如果鼠标落在或达到最大测试时间（任意地设定在120秒），没有S上的测试小光伏棒。用于统计目的的分配，分配小鼠下降，特别棒和后续的120，因为它假定他们会再次下跌，从较窄的杆。如果鼠标变成倒挂，也注意到这一事件，成功的过境以及方向，它必须保持直立杆。否则，分配给它的最高的得分。取出鼠标杆或跌倒后到达终点。 经过测试，在一杆鼠标回到家里笼休息，而测试另一个鼠标。然后将其放在下一个较小尺寸的杆，并再次以同样的方式进行测试。停止测试，如果超过5秒后，鼠标杆脱落。如果它掉下来了，在不到5秒替换它，并允许另一种尝试（如5秒内下降可能是由于被实验者故障配售），最多三项试验，并使用最好的结果。 2.4双杠将鼠标在t他中心的两间酒吧，其纵轴垂直的酒吧，两个前爪应该是在一个酒吧，其他酒吧的两个后爪。杆数是类似于静态棒的。采取两种措施：所花费的时间，直至鼠标的方向90°至开始位置，其后到达的时间，直到它的端部支撑。如果鼠标变成倒挂，也注意到此事件。至于静态棒测试，必须达到的方向和过境不翻转，否则被分配的最大时间。合理健康的小鼠，这是罕见的下跌，在不到5秒，如果这确实发生，它可能是由于配售不佳。因此丢弃的结果，并重新测试鼠标。

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M., Casteels, C., Coun, F., Ni, Y., Van Laere, K., Baekelandt, V. <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></ol>

作者没有利益冲突的披露。

行为显明行动和行动需要运动技能，包括身体的协调。在历史上的第一个实验动物行为之一的，由科林·斯图尔特，在美国11克拉克大学的博士生。他测量老鼠的活动，把它们放入一个鼓，数算转身的时候。这种早期的装置是可能的运行车轮和本发明使用的旋转试验，，尽管的斯图尔特装置中的大鼠滚筒内部的，这是驱动的大鼠的运动，而在rotarod动物行走的外侧上的前身电机驱动的旋转鼓或杆。 原来由邓纳姆和宫（1957年）7杆一组的速度，但如今加速版本（琼斯和罗伯茨1968年）9可用。设定的速度小鼠旋转的缺点是与协调性差，有些动物会脱落开始，而那些留在测试将很快开始测量耐力，而不是本身的协调。将所得的双峰分布的数据点可以足够异常排除参数分析。当用于加速小鼠旋转，尤其是当它被加速，仅当一组条件，如保持为10秒，来实现，数据分配是更可能是高斯的。可以调整的速率加速，使电机不疲劳性能中扮演一个重要的角色;明显疲劳后在小鼠的Rotarod上的时间只需2-3分钟，一个鼠标是不大可能的。 “字符串测试”，“衣架测试”或单杠措施前肢的力量和协调性。两个绰号报价是指其发展;原本是一个字符串，但作者指出，性能很大程度上取决于如何绷紧字符串，因此未来的实验者用一根金属棒，往往是一个衣架。我们已经发现，鼠标的能力夹住酒吧是它的直径成反比，标准使用2毫米的酒吧。 （见朱庇特的协议“测量老鼠的实力”）的极细的金属丝水壶毛皮/规模集热器允许在重量测试最强的动物，以确保他们能够举起超过自身体重的这样一个良好的抓地力。但是，使用直径较大的水平条可以是一个优势，因为测试的灵敏度提高了，只有2毫米bar最正常的C57BL / 6小鼠达到上限的性能。因此，我们最近采用了一个三杆装置，可以随时互换的酒吧，因为他们到合适的支撑柱顶部切成缺口。张力（尤其重要的是要刚性2毫米bar）下举行的酒吧，因为它们只适合的缺口支撑小幅一起绘制（如长弓，并将其字符串）（ 见图3）。 经常被用来静态杆或横梁衡量协调，但额外的动机总是用说服的动物穿越杆。他们协商的水平梁，以达到理想的目标，如家笼，从明亮的光线下避难，或食物（如果他们已经饿了）。然而，在调查的过程中，这种类型的测试中，我发现，小鼠会自发地转身放在“开放”悬臂杆结束时，然后遍历它，直到他们达到支持的结束。这种技术消除了需要培训（必要时在前面提到的测试变体）升高的杆的端部被放置在靠近鼠标的先天反应是试图达到的支撑端。当然，这不，消除动机的转变可能会影响性能的问题，这始终是一种可能性，这种类型的程序。 目前多用于纸板隧道家笼环境富集，但他们的劣势在目前情况下是有利的，动物难免会被实行平衡缸，可以摇晃约其长轴，模仿必要的技能，以平衡对小鼠旋转。因此家笼富集可能掩盖的行为表型，一个例子是亨廷顿疾病表型富集14的改良。动物也经常步行沿长轴这些隧道，行走时升高静态梁。因此，我构建了一个对双杠升高，并放置在中间的老鼠与酒吧和另一方面，他们从来没有遇到过的情况后爪的前爪上。小鼠到达终点支持双杠，而其余的直立通过测试。有些类似的设备是一个系列鼠笼放置在一个双杠，以评估发展痒病引起的运动功能障碍13。 现在，我们已经测试几百在牛津大学的电机运转的小鼠。这些曾接受各种治疗，包括药物，或被动转移组织（浆）从人类，但大多是转基因动物。有些动物更加减值协调任务，强度比别人反之亦然。一些小鼠受损小鼠在Rotarod比静态基材如静态杆和双杠，反之亦然。所以本文随附的朱庇特的协议“在小鼠强度测量”）描述了如何表征的方式，不同的治疗方法和的突变差异可以影响小鼠的运动功能。除小鼠在Rotarod外此描述的仪器简单，价格便宜，特别适合没有专门的运动技能测试实验室。对于判罚尺度的步态分析，的时装表演系统是一个更为复杂的系统比旧的方法，如说服鼠标穿行墨浴，然后下一张纸，但它是一个expensivê片设备。 所有这些测试旨在衡量协调，而不是实力，但显然是一个最低程度的肌张力是必要的仅仅为鼠标使用它的四肢。强度评估，倒屏程序10更适合，或更梯度测量是必需的，的重量测试或自动化的酒吧拉当量（朱庇特的协议“测量强度在小鼠中描述”）。唯一的例外是单杠测试，这既需要力量和协调成功的表现。

measuring motor coordination in mice

正越来越多地被用于小鼠在行为神经科学，在很大程度上取代大鼠行为主义的首选动物。之前的行为，如情绪性或认知的方面可以进行评估，但是，这是至关重要的，以确定是否允许电机的能力， 例如，突变体或损伤小鼠这样的评估。一个迷宫的任务，需要强度和协调，如Morris水迷宫，性能上可能受损，鼠标，由电动机，而不是认知障碍的，所以有必要选择性地解剖后者从前者。例如，感觉障碍引起的NMDA拮抗剂已显示损害水迷宫性能2。 动作协调历来被评定在小鼠和大鼠的旋转试验，在该动物被放置在一个水平杆，其长轴旋转;动物必须向前走，保持直立，不会脱落。这两个设定的速度和加速版本的rotarod都可用。 其他三个测试在这篇文章中（单杠，静电棒和双杠）静态设备上的所有措施的协调。单杠还需要足够的性能实力，尤其是前肢鼠标最初只是用前爪交手吧。 成年大鼠不执行测试，如静态杆和双杠（个人意见）;他们比老鼠出现良好的协调。我只测试雄性大鼠，然而，雄性小鼠似乎比女性一般不很好地协调。老鼠似乎有较高的强度重量比的大鼠相比，拉丁名， 小家鼠 ，似乎完全合适。 rotarod的脚的故障测试12或猫步（Noldus）15装置的变化一般是用来评估大鼠运动协调。

预期结果静态杆原本是用来监测亨廷顿氏病的进展，突变小鼠，带或不带环境富集在他们的家笼14。铀浓缩计划大大推迟了上杆的延迟在起病后爪紧紧抱住它，一般的神经功能缺损的迹象，这是平行运动功能障碍的发病。 感染瘙痒病的小鼠表现出减少在单杠性能从第16周后喷射，在大鼠旋转运动检测从第18周，和静态棒从至 ​​少19周，8（较早的测试没有执行）。 KATP通道亚基的Kir6.2的KO小鼠受损的静态杆和单杠，，但不的rotarod 4。海马病变有任何措施不会影响运动能力5，但129S2/Sv小鼠受损相对C57BL / 6株大鼠旋转运动和静态棒3。 C57BL/10的小鼠也有较差的电机的性能比C57BL / 6小鼠6，虽然没有作为129S2/Sv老鼠到相同的程度。 <img alt="图1" src="/files/ftp_upload/2609/2609fig1.jpg" /> 图1。小鼠旋转。 <img alt="图2" src="/files/ftp_upload/2609/2609fig2.jpg" /> 图2。详细滚花杆的小鼠在Rotarod的。 <img alt="图3" src="/files/ftp_upload/2609/2609fig3.jpg" /> 图3。三联单杠酒吧举行的槽口中，是基本的2毫米1;上涨之下（仅用于演示目的）是4和6毫米的酒吧。在实践中，在同一时间只有一个酒吧开槽成的缺口支撑。 <imG ALT =“图4”SRC =“/ files/ftp_upload/2609/2609fig4.jpg”/&gt; 图4。静态杆穿越鼠标。 <img alt="图5" src="/files/ftp_upload/2609/2609fig5.jpg" /> 图5。鼠标在双杠上。填充已被省略仅用于演示目的。

Watch this Scientific Journal Video about Measuring Motor Coordination in Mice at JoVE.com

Measuring Motor Coordination in Mice

议定书两个电机成立协调任务，加快大鼠旋转运动和单杠，还有两个最近在牛津大学开发的测试，静态棒和双杠。这些测试可以检测运动损伤潜在的兴趣在自己的权利，以及作为可能的变量在其他领域的行为测试。

测量小鼠运动协调

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such ...

Research

JoVE Journal

Neuroscience

99.4K 次观看.  University of Oxford. 议定书两个电机成立协调任务，加快大鼠旋转运动和单杠，还有两个最近在牛津大学开发的测试，静态棒和双杠。这些测试可以检测运动损伤潜在的兴趣在自己的权利，以及作为可能的变量在其他领域的行为测试。

视频: Measuring Motor Coordination in Mice

Assessment of Motor Balance and Coordination in Mice using the Balance Beam

Brain injury, genetic manipulations, and pharmacological treatments can result in alterations of motor skills in mice. Fine motor coordination and balance can be assessed by the beam walking assay. The goal of this test is for the mouse to stay upright and walk across an elevated narrow beam to a safe platform. This test takes place over 3 consecutive days: 2 days of training and 1 day of testing. Performance on the beam is quantified by measuring the time it takes for the mouse to traverse the beam and the number of paw slips that occur in the process. Here we report the protocol used in our laboratory, and representative results from a cohort of C57BL/6 mice. This task is particularly useful for detecting subtle deficits in motor skills and balance that may not be detected by other motor tests, such as the Rotarod.

Deficits in fine motor coordination can be assessed with the balance beam test. Performance on the beam is quantified by the speed at which the beam is traversed and the number of times the mouse slips on the beam.

在小鼠体内的汽车平衡和协调的评估使用的平衡木

Brain injury, genetic manipulations, and pharmacological treatments can result in alterations of motor skills in mice.  Fine motor coordination and ...

科研

神经科学

Measuring Circadian and Acute Light Responses in Mice using Wheel Running Activity

Circadian rhythms are physiological functions that cycle over a period of approximately 24 hours (circadian- circa: approximate and diem: day)1, 2. They are responsible for timing our sleep/wake cycles and hormone secretion. Since this timing is not precisely 24-hours, it is synchronized to the solar day by light input. This is accomplished via photic input from the retina to the suprachiasmatic nucleus (SCN) which serves as the master pacemaker synchronizing peripheral clocks in other regions of the brain and peripheral tissues to the environmental light dark cycle3-7. The alignment of rhythms to this environmental light dark cycle organizes particular physiological events to the correct temporal niche, which is crucial for survival8. For example, mice sleep during the day and are active at night. This ability to consolidate activity to either the light or dark portion of the day is referred to as circadian photoentrainment and requires light input to the circadian clock9. Activity of mice at night is robust particularly in the presence of a running wheel. Measuring this behavior is a minimally invasive method that can be used to evaluate the functionality of the circadian system as well as light input to this system. Methods that will covered here are used to examine the circadian clock, light input to this system, as well as the direct influence of light on wheel running behavior.

This article will review methods that can be used to determine circadian function and light responsiveness in mice.

测量小鼠使用车轮运行活动的昼夜和急性对光反应

Circadian rhythms are physiological functions that cycle over a period of approximately 24 hours (circadian- circa: approximate and diem: day)1, 2.  They ...

Extracellularly Identifying Motor Neurons for a Muscle Motor Pool in Aplysia californica

In animals with large identified neurons (e.g. mollusks), analysis of motor pools is done using intracellular techniques1,2,3,4. Recently, we developed a technique to extracellularly stimulate and record individual neurons in Aplysia californica5. We now describe a protocol for using this technique to uniquely identify and characterize motor neurons within a motor pool.
This extracellular technique has advantages. First, extracellular electrodes can stimulate and record neurons through the sheath5, so it does not need to be removed. Thus, neurons will be healthier in extracellular experiments than in intracellular ones. Second, if ganglia are rotated by appropriate pinning of the sheath, extracellular electrodes can access neurons on both sides of the ganglion, which makes it easier and more efficient to identify multiple neurons in the same preparation. Third, extracellular electrodes do not need to penetrate cells, and thus can be easily moved back and forth among neurons, causing less damage to them. This is especially useful when one tries to record multiple neurons during repeating motor patterns that may only persist for minutes. Fourth, extracellular electrodes are more flexible than intracellular ones during muscle movements. Intracellular electrodes may pull out and damage neurons during muscle contractions. In contrast, since extracellular electrodes are gently pressed onto the sheath above neurons, they usually stay above the same neuron during muscle contractions, and thus can be used in more intact preparations.
To uniquely identify motor neurons for a motor pool (in particular, the I1/I3 muscle in Aplysia) using extracellular electrodes, one can use features that do not require intracellular measurements as criteria: soma size and location, axonal projection, and muscle innervation4,6,7. For the particular motor pool used to illustrate the technique, we recorded from buccal nerves 2 and 3 to measure axonal projections, and measured the contraction forces of the I1/I3 muscle to determine the pattern of muscle innervation for the individual motor neurons.
We demonstrate the complete process of first identifying motor neurons using muscle innervation, then characterizing their timing during motor patterns, creating a simplified diagnostic method for rapid identification. The simplified and more rapid diagnostic method is superior for more intact preparations, e.g. in the suspended buccal mass preparation8 or in vivo9. This process can also be applied in other motor pools10,11,12 in Aplysia or in other animal systems2,3,13,14.

Extracellularly Identifying Motor Neurons for a Muscle Motor Pool in Aplysia californica

In animals with large identified neurons (e.g. mollusks), analysis of motor pools is done using intracellular techniques1,2,3,4. Recently, we developed a technique to extracellularly stimulate and record individual neurons in Aplysia californica5. We now describe a protocol for using this technique to uniquely identify and characterize motor neurons within a motor pool.

运动神经元细胞外识别的肌肉运动游泳池<em&gt;海兔夜蛾</em

In animals with large  identified neurons (e.g. mollusks), analysis of motor pools is done using  intracellular techniques1,2,3,4. Recently,  we developed ...

Measuring Changes in Tactile Sensitivity in the Hind Paw of Mice Using an Electronic von Frey Apparatus

Measuring inflammation-induced changes in thresholds of hind paw withdrawal from mechanical pressure is a useful technique to assess changes in pain perception in rodents. Withdrawal thresholds can be measured first at baseline and then following drug, venom, injury, allergen,&#160;or otherwise evoked inflammation by applying an accurate force on very specific areas of the skin. An electronic von Frey apparatus allows precise assessment of mouse hind paw withdrawal thresholds that are not limited by the available filament sizes in contrast to classical von Frey measurements. The ease and rapidity of measurements allow for incorporation of assessment of tactile sensitivity outcomes in diverse models of rapid-onset inflammatory and neuropathic pain as multiple measurements can be taken within a short time period. Experimental measurements for individual rodent subjects can be internally controlled against individual baseline responses and exclusion criteria easily established to standardize baseline responses within and across experimental groups. Thus, measurements using an electronic von Frey apparatus represent a useful modification of the well-established classical von Frey filament-based assays for rodent mechanical allodynia that may also be applied to other nonhuman mammalian models.

Electronic von Frey measurements are an effective and improved alternative to classical von Frey filaments, as they allow rapid and precise measurement of changes in rodent mechanical withdrawal thresholds to continuously applied pressure stimuli before and after an inflammatory event.

Measuring inflammation-induced changes in thresholds of hind paw withdrawal from mechanical pressure is a useful technique to assess changes in pain ...

Measuring Spinal Presynaptic Inhibition in Mice By Dorsal Root Potential Recording In Vivo

Presynaptic inhibition is one of the most powerful inhibitory mechanisms in the spinal cord. The underlying physiological mechanism is a depolarization of primary afferent fibers mediated by GABAergic axo-axonal synapses (primary afferent depolarization). The strength of primary afferent depolarization can be measured by recording of volume-conducted potentials at the dorsal root (dorsal root potentials, DRP). Pathological changes of presynaptic inhibition are crucial in the abnormal central processing of certain pain conditions and in some disorders of motor hyperexcitability. Here, we describe a method of recording DRP in vivo in mice. The preparation of spinal cord dorsal roots in the anesthetized animal and the recording procedure using suction electrodes are explained. This method allows measuring GABAergic DRP and thereby estimating spinal presynaptic inhibition in the living mouse. In combination with transgenic mouse models, DRP recording may serve as a powerful tool to investigate disease-associated spinal pathophysiology. In vivo recording has several advantages compared to ex vivo isolated spinal cord preparations, e.g. the possibility of simultaneous recording or manipulation of supraspinal networks and induction of DRP by stimulation of peripheral nerves.

Measuring Spinal Presynaptic Inhibition in Mice By Dorsal Root Potential Recording In Vivo

GABAergic presynaptic inhibition is a powerful inhibitory mechanism in the spinal cord important for motor and sensory signal integration in spinal cord networks. Underlying primary afferent depolarization can be measured by recording of dorsal root potentials (DRP). Here we demonstrate a method of in vivo recording of DRP in mice.

测量脊髓突触前抑制小鼠背根电位记录<em&gt;在体内</em

Presynaptic inhibition is one of the most powerful inhibitory mechanisms in the spinal cord. The underlying physiological mechanism is a depolarization of ...

The Neuromuscular Junction: Measuring Synapse Size, Fragmentation and Changes in Synaptic Protein Density Using Confocal Fluorescence Microscopy

The neuromuscular junction (NMJ) is the large, cholinergic relay synapse through which mammalian motor neurons control voluntary muscle contraction. Structural changes at the NMJ can result in neurotransmission failure, resulting in weakness, atrophy and even death of the muscle fiber. Many studies have investigated how genetic modifications or disease can alter the structure of the mouse NMJ. Unfortunately, it can be difficult to directly compare findings from these studies because they often employed different parameters and analytical methods. Three protocols are described here. The first uses maximum intensity projection confocal images to measure the area of acetylcholine receptor (AChR)-rich postsynaptic membrane domains at the endplate and the area of synaptic vesicle staining in the overlying presynaptic nerve terminal. The second protocol compares the relative intensities of immunostaining for synaptic proteins in the postsynaptic membrane. The third protocol uses Fluorescence Resonance Energy Transfer (FRET) to detect changes in the packing of postsynaptic AChRs at the endplate. The protocols have been developed and refined over a series of studies. Factors that influence the quality and consistency of results are discussed and normative data are provided for NMJs in healthy young adult mice.

The neuromuscular junction (NMJ) is altered in a variety of conditions that can sometimes culminate in synaptic failure. This report describes fluorescence microscope-based methods to quantify such structural changes.

神经肌肉接头处：测量突触大小，碎片和改变突触蛋白密度利用共聚焦荧光显微镜

The neuromuscular junction (NMJ) is the large, cholinergic relay synapse through which mammalian motor neurons control voluntary muscle contraction. ...

Polygraphic Recording Procedure for Measuring Sleep in Mice

Recording of the epidural electroencephalogram (EEG) and electromyogram (EMG) in small animals, like mice and rats, has been pivotal to study the homeodynamics and circuitry of sleep-wake regulation. In many laboratories, a cable-based sleep recording system is used to monitor the EEG and EMG in freely behaving mice in combination with computer software for automatic scoring of the vigilance states on the basis of power spectrum analysis of EEG data. A description of this system is detailed herein. Steel screws are implanted over the frontal cortical area and the parietal area of 1 hemisphere for monitoring EEG signals. In addition, EMG activity is monitored by the bilateral placement of wires in both neck muscles. Non-rapid eye movement (Non-REM; NREM) sleep is characterized by large, slow brain waves with delta activity below 4 Hz in the EEG, whereas a shift from low-frequency delta activity to a rapid low-voltage EEG in the theta range between 6 and 10 Hz can be observed at the transition from NREM to REM sleep. By contrast, wakefulness is identified by low- to moderate-voltage brain waves in the EEG trace and significant EMG activity.

The recording of electroencephalogram (EEG) and electromyogram (EMG) in freely behaving mice is a critical step to correlate behavior and physiology with sleep and wakefulness. The experimental protocol described herein provides a cable-based system for acquiring EEG and EMG recordings in mice.

印刷业记录程序在小鼠体内测量睡眠

Recording of the epidural electroencephalogram (EEG) and electromyogram (EMG) in small animals, like mice and rats, has been pivotal to study the ...

Retrograde Neuroanatomical Tracing of Phrenic Motor Neurons in Mice

Phrenic motor neurons are cervical motor neurons originating from C3 to C6 levels in most mammalian species. Axonal projections converge into phrenic nerves innervating the respiratory diaphragm. In spinal cord slices, phrenic motor neurons cannot be identified from other motor neurons on morphological or biochemical criteria. We provide the description of procedures for visualizing phrenic motor neuron cell bodies in mice, following intrapleural injections of cholera toxin subunit beta (CTB) conjugated to a fluorophore. This fluorescent neuroanatomical tracer has the ability to be caught up at the diaphragm neuromuscular junction, be carried retrogradely along the phrenic axons and reach the phrenic cell bodies. Two methodological approaches of intrapleural CTB delivery are compared: transdiaphragmatic versus transthoracic injections. Both approaches are successful and result in similar number of CTB-labeled phrenic motor neurons. In conclusion, these techniques can be applied to visualize or quantify the phrenic motor neurons in various experimental studies such as those focused on the diaphragm-phrenic circuitry.

Here, we describe a protocol for identifying phrenic motor neurons in mice after intrapleural delivery of fluorophore conjugated cholera toxin subunit beta. Two techniques are compared to inject the pleural cavity: transdiaphragmatic versus transthoracic approaches.

小鼠膈运动神经元逆行神经解剖学追踪的研究

Phrenic motor neurons are cervical motor neurons originating from C3 to C6 levels in most mammalian species. Axonal projections converge into phrenic ...

Evaluation of Hemisphere Lateralization with Bilateral Local Field Potential Recording in Secondary Motor Cortex of Mice

This article demonstrates complete, detailed procedures for both in vivo bilateral recording and analysis of local field potential (LFP) in the cortical areas of mice, which are useful for evaluating possible laterality deficits, as well as for assessing brain connectivity and coupling of neural network activities in rodents. The pathological mechanisms underlying Alzheimer&#39;s disease (AD), a common neurodegenerative disease, remain largely unknown. Altered brain laterality has been demonstrated in aging people, but whether or not abnormal lateralization is one of the early signs of AD has not been determined. To investigate this, we recorded bilateral LFPs in 3-5-month-old AD model mice, APP/PS1, together with littermate wild type (WT) controls. The LFPs of the left and right secondary motor cortex (M2), specifically in the gamma band, were more synchronized in APP/PS1 mice than in WT controls, suggesting a declined hemispheric asymmetry of bilateral M2 in this AD mouse model. Notably, the recording and data analysis processes are flexible and easy to carry out, and can also be applied to other brain pathways when conducting experiments that focus on neuronal circuits.

We present in vivo electrophysiological recording of the local field potential (LFP) in bilateral secondary motor cortex (M2) of mice, which can be applied to evaluate hemisphere lateralization. The study revealed altered levels of synchronization between the left and right M2 in APP/PS1 mice compared to WT controls.

小鼠二次运动皮质中双边局部场电位记录半球横向化评价

This article demonstrates complete, detailed procedures for both in vivo bilateral recording and analysis of local field potential (LFP) in the cortical ...

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

Compared to the Sciatic Functional Index (SFI), kinematic analysis is a more reliable and sensitive method for performing functional evaluations of sciatic nerve injury rodent models. In this protocol, we describe a novel kinematic analysis method that uses a three-dimensional (3D) motion capture apparatus for functional evaluations using a rat sciatic nerve crush injury model. First, the rat is familiarized with treadmill walking. Markers are then attached to the designated bone landmarks and the rat is made to walk on the treadmill at the desired speed. Meanwhile, the posterior limb movements of the rat are recorded using four cameras. Depending on the software used, marker tracings are created using both automatic and manual modes and the desired data are produced after subtle adjustments. This method of kinematic analysis, which uses a 3D motion capture apparatus, offers numerous advantages, including superior precision and accuracy. Many more parameters can be investigated during the comprehensive functional evaluations. This method has several shortcomings that require consideration: The system is expensive, can be complicated to operate, and may produce data deviations due to skin shifting. Nevertheless, kinematic analysis using a 3D motion capture apparatus is useful for performing functional anterior and posterior limb evaluations. In the future, this method may become increasingly useful for generating accurate assessments of various traumas and diseases.

We introduce a kinematic analysis method that uses a three-dimensional motion capture apparatus containing four cameras and data processing software for performing functional evaluations during fundamental research involving rodent models.

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 ...