这项研究是由来自弗吉尼亚州的联邦卫生研究委员会补助经费支持。

为便于说明的协议已被划分成两部分:1）实验步骤和2）的自动分析。所有的动物都根据指南实验动物15的护理和使用治疗，所有手续由机构动物护理和使用委员会批准。 1.实验步骤<ol><li>设置保持架，这是一个黑色的无反射有机玻璃矩形箱（52×25×23 毫升 ）分成三个室:社会，非社会，和中性中间隔间（ 图1）。放置在会议期间在各端隔室的倒置丝杯I和II（下文讨论）和会话第二期间容纳所述刺激小鼠。 </li><li>设置摄像机，所述笼填充的视场的方式。由于照明可以用鼠标行为干预，维持在笼子里面不超过3.5勒克斯的照度，从间接光从未来白炽灯泡和在向上方向上的笼底测量。也使用红外光源，小鼠无法感知但相机可检测。记录所有电影720×480标清。 </li><li>录制后图像处理的空笼（称为"参照系"）的一帧，并保存为灰度图像。 </li><li>第一节:介绍测试鼠标放到笼子里和波拱手笼，表明会话的开始（比较自动分析）。影片记录了10分钟。在10分钟结束时，把鼠标放回原来的笼子里。 </li><li>议题二:把鼠标刺激倒杯之一的（平衡）。 </li><li>使用"社会左"或"社会权利"按钮，选择刺激的鼠标是否是在左侧还是在右侧的倒杯的倒杯。 注:与刺激鼠标倒杯被称为"社会的杯具"，而其他杯"非社会茶杯"。其中社会杯位于被称为社会隔室的隔室，与非社会杯隔室是非社会隔室，中间室是"中性隔室"。 </li><li>引进试验老鼠入笼。波拱手笼指示会话的开始。影片记录了10分钟。重要的是，保持社交性设备移除老鼠倒杯开始第三场之前。 </li><li>议题三:删除倒杯，松开小鼠和允许测试和刺激老鼠相互自由交互。波拱手笼指示会话的开始。影片记录了10分钟。在10分钟结束时，把老鼠都回到各自的笼子里。 </li></ol> 2.自动分析注:在第2的命令经由图形用户界面发出（参见图2），该CONtrols我们的软件。在这里我们讲解一步一步的处理，我们的软件允许;该软件还能够在无用户输入执行批处理模式的分析。 <ol><li>对于每部电影，请执行以下处理步骤: <ol><li>负载电影（点击Load按钮）。 </li><li>选择笼构成，它告诉分析程序笼边界，隔室的边界，和杯位置的位置。选择四个保存的配置中的一种或手动调整所有坐标。 注:配置通常不改变从一个实验到下一个，因此可以重复使用。 </li><li>为电影的每一帧，执行以下的处理步骤（2.1.3.1-2.1.3.6）。 <ol><li>转换帧通过点击"灰度"按钮灰度。 </li><li>通过点击"减去"按钮，数字减去灰度参考框架（见步骤1.3）。注意:该步骤的目的是使像素VA框架非常接近的梅毒到处为零，除了鼠标的位置。 </li><li>通过设置低于某个阈值电平为黑色的所有像素二值化的帧，高于该水平的白色所有像素（点击"二值化"按钮）。 注意:我们的想法是，由于鼠标象素通常具有较高的值比剩余像素，鼠标像素将变成白色，所有其他像素黑色。最佳阈值电平取决于照明条件，并且可以通过在整个电影评估鼠标形状来确定。如果选择阈值太低，则鼠标出现过大，以及不属于鼠标像素可被着色为白色。如果阈值太高，鼠标变小，并且可以分裂成断开件。在记录的电影，最佳阈值总是20和35之间（从0到255的范围）。 </li><li>侵蚀13二进制图像e时代，那么13扩张2E次，然后ERO德再次e时代，按下按钮"EROD /语"。 注意:目的此过程是删除尾部。对于已使用的分辨率（720×480），E = 3在所有情况下工作。 注:该软件的手在画面中存在自动检查（动画的录制过程中，手都通过图片摆手示意会话的开始）。如果手存在，就会有在二值化的帧异常许多白色像素，因为手是比鼠标大得多。一只手的存在的条件是，有在二值化的帧超过10000白色像素。该软件I，II和III，通过检测其手部动作发生的帧的范围确定会议的开端。 </li></ol></li></ol></li><li>在会话I和II，继续如下: <ol><li>检测组白色像素的最大连接组件（点击"LCOMP"）。注:目的ØF该步骤是为了消除该不属于鼠标， 例如 ，引起刺激小鼠的会话第二运动白色像素。最大的连接组件是在几乎所有帧的鼠标。 </li><li>通过点击"查找完"确定了两个最遥远的白色像素（称为鼠标的"目的"）。 注意:软件建立的两端， 即的连续性，对于每个在第一帧的端部到在它们所属的后续帧的端部。总的想法是，由于帧以高速率记录中，端部可以不远处一帧行进到下一个，使得用于从一帧到下一的端部的正确延续是该距离的总和最小化两端移动。见讨论更多细节。 </li><li>确定哪些鼠标到底是鼻子。 <ol><li>确定的方向，其中通过观察一个移动鼠标如何重心（COG）从帧变为帧（焦炉煤气的动议是所有小鼠像素的总和的运动）。 注意:一组象素P 1的COG，...，坐标P N×1，...，×n个被定义为: <img alt="式（1）" src="/files/ftp_upload/52508/52508eq1.jpg" /> 鼠标通常向前（ 即往自己的鼻子）。在几百帧的部分，总是有很高可信度的头部/尾部的检测。 注意:软件自动确定作为所检测到结束鼠标移动;为此被识别为鼻和标有在电影的红色圆圈。当检测到的鼻子被标记在每一个帧中的红色圆圈，用户可以检查该检测的质量。 </li></ol></li></ol></li><li>在会议第三，继续如下: <ol><li>检测两个最大的白色像素组件。 <ol><li>如果两者都为上午可接受大小的乌斯和其重心（COGS）是与前一帧的鼠标COGS，考虑这两个部件的两只小鼠兼容。 </li><li>考虑鼠标尺寸上可接受的，如果他们不超过20％，因为前一帧改变，并且考虑连续两帧为兼容的嵌齿，如果它们的差值不大于鼠标可以在帧之间的时间移动的距离越大（在此距离如下从最大鼠标速度，这可以通过分析多个单独的小鼠的运动被确定为一个实验系列的制剂的一部分（如在会话I和II）。 注意:如果最大的两个组成部分是一个鼠标不能接受尺寸或嵌齿是从那些在先前帧过于不同，直到最大的两个组成部分是可接受的大小的软件自动增加二值化阈值，并且它们的齿轮与兼容那些先前帧。 </li></ol></li><li>点击测试鼠标第一帧。在使用2.3.1分析会议III的每一帧识别测试和刺激鼠标。 </li><li>确定会议III，其中一个鼠标社会接近其他的帧。注意:软件实现此步骤通过自动检查的小鼠之间的距离小于小鼠的典型相互作用距离（我们使用2个厘米）。 </li><li>当软件播放影片与所检测的方式向用户的一个短段，点击一个按钮，以指示该方法是否符合由测​​试小鼠，社交回避由测试鼠标，或既不是社会的方法。 注意:步骤2.3.4能，如果它是已知的鼠标是测试鼠标自动化。由于当小鼠太靠近彼此此信息丢失（或爬在彼此的顶部），而不是因为我们还没有发现一种方法来标记的小鼠中，可以可靠地通过我们的软件进行检测并执行的方式不会影响他们的行为，我们已经恢复了人工分类现在。 </li></ol></li><li>要使用Load按钮评估处理电影，电影负荷。 <ol><li>点击开始会话1显示会话1的第一帧（或开始第二节，显示会话2的第一帧）。 </li><li>点击播放/停止播放和停止影片。 注:处理影片将显示鼠标的头部和尾部标有不同颜色的圆圈。显示在屏幕上的所有社交措施和更新的影片播放。 </li><li>通过点击"导出"导出社交性Excel格式的数据。 </li><li>点击"编译数据"编译文件夹中的所有影片中所有的数据到一个单一的Excel文件。 </li></ol></li></ol>

<ol><li>Brodkin, E. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((BALB%2Fc+mice%3A+Low+sociability+and+other+phenotypes+that+may+be+relevant+to+autism%5BTitle%5D)+AND+%22Behavioural+Brain+Research%22%5BJournal%5D)">BALB/c mice: Low sociability and other phenotypes that may be relevant to autism.</a> Behavioural Brain Research. 176, 53-65 (2007).</li>
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<li>Babineau, B. A., Yang, M., Berman, R. F., Crawley, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Low+home+cage+social+behaviors+in+BTBR+T%2Btf%2FJ+mice+during+juvenile+development%5BTitle%5D)+AND+%22Physiology+%26+Behavior%22%5BJournal%5D)">Low home cage social behaviors in BTBR T+tf/J mice during juvenile development.</a> Physiology & Behavior. 114-115, 49-54 (2013).</li>
<li>Benson, A. D., Burket, J. A., Deutsch, S. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((BALB%2Fc+mice+treated+with+d-cycloserine+arouse+increased+social+interest+in+conspecifics%5BTitle%5D)+AND+%22Brain+Research+Bulletin%22%5BJournal%5D)">BALB/c mice treated with d-cycloserine arouse increased social interest in conspecifics.</a> Brain Research Bulletin. 99, 95-99 (2013).</li>
<li>Burket, J. A., Benson, A. D., Tang, A. H., Deutsch, S. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Rapamycin+improves+sociability+in+the+BTBR+T%28%2B%29Itpr3%28tf%29%2FJ+mouse+model+of+autism+spectrum+disorders%5BTitle%5D)+AND+%22Brain+Research+Bulletin%22%5BJournal%5D)">Rapamycin improves sociability in the BTBR T(+)Itpr3(tf)/J mouse model of autism spectrum disorders.</a> Brain Research Bulletin. 100, 70-75 (2014).</li>
<li>Burket, J. A., Benson, A. D., Tang, A. H., Deutsch, S. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((D-Cycloserine+improves+sociability+in+the+BTBR+T%2B+Itpr3tf%2FJ+mouse+model+of+autism+spectrum+disorders+with+altered+Ras%2FRaf%2FERK1%2F2+signaling%5BTitle%5D)+AND+%22Brain+Research+Bulletin%22%5BJournal%5D)">D-Cycloserine improves sociability in the BTBR T+ Itpr3tf/J mouse model of autism spectrum disorders with altered Ras/Raf/ERK1/2 signaling.</a> Brain Research Bulletin. 96, 62-70 (2013).</li>
<li>Burket, J. A., Herndon, A. L., Deutsch, S. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Locomotor+activity+of+the+genetically+inbred+BALB%2Fc+mouse+strain+is+suppressed+by+a+socially+salient+stimulus%5BTitle%5D)+AND+%22Brain+Research+Bulletin%22%5BJournal%5D)">Locomotor activity of the genetically inbred BALB/c mouse strain is suppressed by a socially salient stimulus.</a> Brain Research Bulletin. 83, 255-256 (2010).</li>
<li>Deutsch, S. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((D-cycloserine+improves+sociability+and+spontaneous+typic+behaviors+in+4-week+old+mice%5BTitle%5D)+AND+%22Brain+Research%22%5BJournal%5D)">D-cycloserine improves sociability and spontaneous typic behaviors in 4-week old mice.</a> Brain Research. 1439, 96-107 (2012).</li>
<li>Jacome, L. F., Burket, J. A., Herndon, A. L., Deutsch, S. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Genetically+inbred+BALB%2Fc+mice+differ+from+outbred+Swiss+Webster+mice+on+discrete+measures+of+sociability%3A+relevance+to+a+genetic+mouse+model+of+autism+spectrum+disorders%5BTitle%5D)+AND+%22Autism+Research%22%5BJournal%5D)">Genetically inbred BALB/c mice differ from outbred Swiss Webster mice on discrete measures of sociability: relevance to a genetic mouse model of autism spectrum disorders.</a> Autism Research. 4, 393-400 (2011).</li>
<li>Jacome, L. F., Burket, J. A., Herndon, A. L., Cannon, W. R., Deutsch, S. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((D-serine+improves+dimensions+of+the+sociability+deficit+of+the+genetically-inbred+BALB%2Fc+mouse+strain%5BTitle%5D)+AND+%22Brain+Research+Bulletin%22%5BJournal%5D)">D-serine improves dimensions of the sociability deficit of the genetically-inbred BALB/c mouse strain.</a> Brain Research Bulletin. 84, 12-16 (2011).</li>
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<li>National Research Council. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=%22Guide+for+the+Care+and+Use+of+Laboratory+Animals%22">Guide for the Care and Use of Laboratory Animals</a>. , Eighth Edition, The National Academic Press. (2011).</li>
</ol>

作者什么都没有透露。

通过结合偷拍小鼠和自动电影的分析，我们已经创建了一个经济实惠，可靠的高通量筛选技术。 自动分析的准确度为100多部电影手工分析的结果进行比较。在帧的绝大多数帧（&gt; 99％），鼠标的鼻部被识别以良好的精度。大部分的（少数）误检测的对社交性措施没有效果，因此，电影的80％左右，不需要任何校正。 在那些确实包含错误检测，是迄今为止...

受损社交性是在一系列神经发育障碍，包括孤独症谱系障碍（自闭症）1,2的主要症状域之一。这严重限制了患者的发展关系的能力，并极大地影响了病人，家属和护理人员2,3。 在为了更好地理解受损社交性背后的机理和开发治疗，几种小鼠模型进行了研究1,4-9。一个这样的菌株是BALB / c小鼠，表现出显着低社交性，类似于患者ASD5 8-12可见特征的近交系。例如，BALB / c小鼠显示社会的方法和通过刺激老鼠（ 例如，社交回避）超过瑞士韦伯斯特比较10株对社会的办法通常是一个否定的答复较低的频率。 为了测试提出的治疗是否有效实现了在增加社交E，需要客观的措施。一种广泛接受的框架是分析在具有三个隔室的标准化装置社交性（ 见图1）。 左，右室都包含一个倒线杯里面有一个突出的社会刺激鼠标。测试小鼠被释放到装置的10分钟三个区间（会话Ⅰ-Ⅲ）;会话我评估测试小鼠的运动活性，而会话II和III评估测试小鼠的社交性的不同方面。 在会话我，测试鼠标使其适应该装置。在此会话，测量该测试鼠标使得不同区室之间的转变的数目。重要的是，在标准的三隔室装置得到社交性措施取决于运动活性，并降低了运动活性可混淆的SOCI解释能力数据。 第二场会议，测试鼠标自由移动，并刺激鼠标倒置一只杯子之下（平衡）。刺激小鼠4周龄远系繁殖雄性ICR小鼠，年龄和性别相匹配的测试小鼠。刺激小鼠被封闭在指定为社会隔室侧的倒置的杯，和一个空的倒置的杯被放置在指定为非社会隔室的一侧。第二场会议，测试鼠标的社交性是通过比较其花费附近的"社会"倒杯（小于2厘米的距离） 与它花费附近的"非社会"倒杯时的时间量化，并通过时间它花费在社交车厢与它在非社会隔间花费的时间。瑞士韦伯斯特比较小鼠花费显著更多的时间在社会隔室和邻近相比BALB / c小鼠，显示没有显著偏爱社会社会倒杯倒杯或社会舱12。 在会话III，测试和刺激小鼠被允许自由互动（倒杯被删除）。车厢之间的社交性措施，刻板行为和过渡的测试和刺激小鼠和分析之间的自由互动的第三10分钟会话中获得可靠。在这份报告中，测试鼠标的社交性通过两个措施量化:1）它使向着刺激鼠标，在一个社会的方式被定义为向着刺激鼠标前进（鼻第一）的社交方式号码，这样的鼻子是从刺激小鼠小于2厘米的距离; 2）由刺激小鼠社会方法忌的数量。测试鼠标被认为是避免方法，如果事实证明或刺激鼠标移开，或者如果它的刺激后鼠标会暂时停止移动（"冻结行为"），已接近测试鼠标。 BALB /c小鼠显示在朝向刺激小鼠和增加的由刺激小鼠社会办法否定答复的号码进行社会的方法数量的减少，相比于瑞士韦伯斯特比较应变10。 行为电影的评价历来手工完成， 即通过（可能以较低的速度），仔细观看电影和激活秒表的鼠标在某车厢或接近倒杯10的一个花费的时间。这种方法需要重复的电影的观看，因此是非常耗时。另外，精度由评价者的反应时间为秒表测量和能力的限制来感知鼠标是否为2cm倒杯内。在最近的工作中，两种化合物（D-丝氨酸和D-环丝氨酸）已被证明有效，在BALB / c小鼠9,10,12恢复社交性。已知的是两种成分MODU迟NMDA受体，这是众所周知的社会行为2,12发挥中心作用。化合物的大阵列已知调节在各种良好定义的方式NMDA受体活性，而对于在ASD治疗的突破的潜力，使这些化合物优先将评价高度期望快速自动方法的加速评估。 下面描述的是用于小鼠社交性的评估开发了一种自动分析过程。它使用鼠标电影的所有帧的图像处理方法来确定鼠标或小鼠的位置。在会话I和II，它进一步检测鼻子的位置，并使用它来计算社交性的措施。在会议第三，它会自动检测所有的方法，但需要人工分类（社会的方式，社交回避，或其他）。

<table><tbody><tr><td>Plexiglas cage</td><td>Norva Plastics, Norfolk, VA</td><td>custom made</td><td>Dimensions and layout described in manuscript,&lt;br/&gt; can be adjusted according to needs.&lt;br/&gt; Use non-reflective plexiglas for to facilitate image processing</td></tr><tr><td>Wire cups</td><td>Kitchen plus</td><td>315</td><td>Use one to house stimulus mouse, one empty</td></tr><tr><td>Video camera</td><td>SONY</td><td>HDR-PJ790</td><td>Can be replaced by any camera with&lt;br/&gt; Comparable specifications</td></tr><tr><td>OpenCV (Image processing library)</td><td>Willow Garage</td><td>N/A</td><td>Any modern image processing library&lt;br/&gt; can be used.</td></tr></tbody></table>

experimental assessment of mouse sociability using an automated image processing approach

鼠标是检验药物设计，增加社交性的首选模式生物。我们提出量化小鼠社交性其中试验小鼠被放置在一个标准化的装置和相关的行为在三个不同的会话评估（称为会话I，II和III）的方法。 该装置具有三个隔室（参见图1），左和右室含有一个倒置的杯，可容纳一个鼠标（称为"刺激小鼠"）。 在会话我，测试小鼠被放置在笼和其移动性的特征在于舱室之间进行转换的数量。第二场会议，刺激老鼠置于倒杯之一的，测试鼠标的社交性是由一次花费近包含封闭刺激鼠标与空倒杯杯的数额量化。在会话III，倒杯卸下了<html> ð两个小鼠自由互动。在会议III测试鼠标的社交性是由它使向着刺激老鼠的社会方式的数量和次数它避免受了刺激鼠标一个社会的办法量化。 影片的自动评估检测测试鼠标，它允许会话I和II的所有描述社交性措施的决心鼻子（会话III，方法被自动识别，但手动分类）。找到鼻，一个空的保持器的图像被数字从影片的每一帧中减去，所得的图像进行二值化，以识别鼠标像素。小鼠尾部被自动删除，剩余的小鼠的两个最远点被确定;这些都是接近机头和机尾的基地。通过分析鼠标的移动和使用的连续性参数，鼻子识别。 <img alt="&lt;/ HTML">"图1"的src ="/文件/ ftp_upload / 52508 / 52508fig1.jpg"/&gt; 图1.社交性的评估期间3届 会议我 （上）:测试鼠标的驯化笼子第二节 （中）:当鼠标刺激被封闭在一个倒杯测试鼠标在笼子里自由移动的第三场 （下图）:两种测试鼠标和鼠标刺激允许自由移动和互动与对方。

图3示出一个行为电影的与空笼一个彩色帧。鼠标还没有被引入笼子里呢。的杯子，（社会和非社会）以及隔室边界的位置重叠。 如图4和用作基准帧的空笼的彩色帧被转换为8位灰度。基准帧被从捕获的电影的每隔一帧中减去。 图5显示从电影会话我在帧中的一个，再次叠加隔间边界和杯子的位置。参照帧是从样本帧，其结果，所谓的差?...

Watch this Scientific Journal Video about Experimental Assessment of Mouse Sociability Using an Automated Image Processing Approach at JoVE.com

Experimental Assessment of Mouse Sociability Using an Automated Image Processing Approach

本协议描述的量化鼠标社交性的方法。因为它们移动和在一个特殊的笼相互作用小鼠录像。电影处理允许社交性具有优良的准确性和可靠性的自动化定量。

使用自动图像处理方法鼠标社交性的实验评估

Mouse is the preferred model organism for testing drugs designed to increase sociability. We present a method to quantify mouse sociability in which the ...

experimental-assessment-mouse-sociability-using-an-automated-image

Research

JoVE Journal

Behavior

8.4K 次观看.  Old Dominion University. 本协议描述的量化鼠标社交性的方法。因为它们移动和在一个特殊的笼相互作用小鼠录像。电影处理允许社交性具有优良的准确性和可靠性的自动化定量。 

视频: Experimental Assessment of Mouse Sociability Using an Automated Image Processing Approach

Automated, Quantitative Cognitive/Behavioral Screening of Mice: For Genetics, Pharmacology, Animal Cognition and Undergraduate Instruction

We describe a high-throughput, high-volume, fully automated, live-in 24/7 behavioral testing system for assessing the effects of genetic and pharmacological manipulations on basic mechanisms of cognition and learning in mice. A standard polypropylene mouse housing tub is connected through an acrylic tube to a standard commercial mouse test box. The test box has 3 hoppers, 2 of which are connected to pellet feeders. All are internally illuminable with an LED and monitored for head entries by infrared (IR) beams. Mice live in the environment, which eliminates handling during screening. They obtain their food during two or more daily feeding periods by performing in operant (instrumental) and Pavlovian (classical) protocols, for which we have written protocol-control software and quasi-real-time data analysis and graphing software. The data analysis and graphing routines are written in a MATLAB-based language created to simplify greatly the analysis of large time-stamped behavioral and physiological event records and to preserve a full data trail from raw data through all intermediate analyses to the published graphs and statistics within a single data structure. The data-analysis code harvests the data several times a day and subjects it to statistical and graphical analyses, which are automatically stored in the &#34;cloud&#34; and on in-lab computers. Thus, the progress of individual mice is visualized and quantified daily. The data-analysis code talks to the protocol-control code, permitting the automated advance from protocol to protocol of individual subjects. The behavioral protocols implemented are matching, autoshaping, timed hopper-switching, risk assessment in timed hopper-switching, impulsivity measurement, and the circadian anticipation of food availability. Open-source protocol-control and data-analysis code makes the addition of new protocols simple. Eight test environments fit in a 48 in x 24 in x 78 in&#160;cabinet; two such cabinets (16 environments) may be controlled by one computer.

Fully automated system for measuring physiologically meaningful properties of the mechanisms mediating spatial localization, temporal localization, duration, rate&#160;and probability estimation, risk assessment, impulsivity, and the accuracy and precision of memory, in order to assess the effects of genetic and pharmacological manipulations on foundational mechanisms of cognition in mice.

小鼠自动化，定量认知/行为筛选：对于遗传学，药理学，动物认知与本科教学

We describe a high-throughput, high-volume, fully automated, live-in 24/7 behavioral testing system for assessing the effects of genetic and ...

科研

行为学

Behavioral Phenotyping of Murine Disease Models with the Integrated Behavioral Station (INBEST)

Due to rapid advances in genetic engineering, small rodents have become the preferred subjects in many disciplines of biomedical research. In studies of chronic CNS disorders, there is an increasing demand for murine models with high validity at the behavioral level. However, multiple pathogenic mechanisms and complex functional deficits often impose challenges to reliably measure and interpret behavior of chronically sick mice. Therefore, the assessment of peripheral pathology and a behavioral profile at several time points using a battery of tests are required. Video-tracking, behavioral spectroscopy, and remote acquisition of physiological measures are emerging technologies that allow for comprehensive, accurate, and unbiased behavioral analysis in a home-base-like setting. This report describes a refined phenotyping protocol, which includes a custom-made monitoring apparatus (Integrated Behavioral Station, INBEST) that focuses on prolonged measurements of basic functional outputs, such as spontaneous activity, food/water intake and motivated behavior in a relatively stress-free environment. Technical and conceptual improvements in INBEST design may further promote reproducibility and standardization of behavioral studies.

Behavioral Phenotyping of Murine Disease Models with the Integrated Behavioral Station INBEST

Prolonged and comprehensive monitoring of mice in a home-cage environment provides a deeper understanding of aberrant behavior in murine models of brain diseases. This paper describes the Integrated Behavioral Station (INBEST) as the key component of contemporary behavioral analysis.

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 ...

Assessment of Social Interaction Behaviors

Social interactions are a fundamental and adaptive component of the biology of numerous species. Social recognition is critical for the structure and stability of the networks and relationships that define societies. For animals, such as mice, recognition of conspecifics may be important for maintaining social hierarchy and for mate choice 1.
A variety of neuropsychiatric disorders are characterized by disruptions in social behavior and social recognition, including depression, autism spectrum disorders, bipolar disorders, obsessive-compulsive disorders, and schizophrenia. Studies of humans as well as animal models (e.g., Drosophila melanogaster, Caenorhabditis elegans, Mus musculus, Rattus norvegicus) have identified genes involved in the regulation of social behavior 2. To assess sociability in animal models, several behavioral tests have been developed (reviewed in 3). Integrative research using animal models and appropriate tests for social behavior may lead to the development of improved treatments for social psychopathologies.
The three-chamber paradigm test known as Crawley's sociability and preference for social novelty protocol has been successfully employed to study social affiliation and social memory in several inbred and mutant mouse lines (e.g. 4-7). The main principle of this test is based on the free choice by a subject mouse to spend time in any of three box's compartments during two experimental sessions, including indirect contact with one or two mice with which it is unfamiliar. To quantitate social tendencies of the experimental mouse, the main tasks are to measure a) the time spent with a novel conspecific and b) preference for a novel vs. a familiar conspecific. Thus, the experimental design of this test allows evaluation of two critical but distinguishable aspects of social behavior, such as social affiliation/motivation, as well as social memory and novelty. "Sociability" in this case is defined as propensity to spend time with another mouse, as compared to time spent alone in an identical but empty chamber 7. "Preference for social novelty" is defined as propensity to spend time with a previously unencountered mouse rather than with a familiar mouse 7. This test provides robust results, which then must be carefully analyzed, interpreted and supported/confirmed by alternative sociability tests. In addition to specific applications, Crawley's sociability test can be included as an important component of general behavioral screen of mutant mice.

Here we describe a detailed protocol for examination of sociability in mice by using Crawley's sociability and preference for social novelty test. We describe the advantages and possible applications for this procedure, including critical details important for correct interpretation of the results.

社会交往行为的评估

Social interactions are a fundamental and adaptive component of the biology of numerous species. Social recognition is critical for the structure and ...

神经科学

Simple and Computer-assisted Olfactory Testing for Mice

Olfaction is highly conserved among species and is required for reproduction and survival.
In humans, olfaction is also one of the senses that is affected with aging and is a strong predictor of neurodegenerative diseases. Thus, olfaction testing is used as a non-invasive diagnostic method to detect neurological deficits early on. In order to understand the mechanisms underlying olfactory network susceptibility, olfactory research in rodents has gained momentum in the past decade.
Here, we present a very simple, time efficient and reproducible olfactory testing method of innate odor perception and sensitivity in mice without the need of any prior food or water restriction. The tests are performed in a familiar environment to the mice, require only the scents and a 2 min session of odorant exposure. The analysis is performed, post-hoc, using computer-assisted commands on ImageJ and can be, therefore, carried out from start to end by one researcher.
This protocol does not require any special hardware or setup and is indicated for any laboratory interested in testing olfactory perception and sensitivity.

We present a simple and unbiased olfactory test in mice. With this protocol olfactory discrimination, preference, avoidance and sensitivity to a novel odor as compared to water can be assessed in single behavioral sessions. This method is indicated for a single experimenter and analysis is based on computer-assisted video processing.

简单和计算机辅助测试嗅觉的小鼠

Olfaction is highly conserved among species and is required for reproduction and survival.
In humans, olfaction is also one of the senses that is affected ...

Recording Mouse Ultrasonic Vocalizations to Evaluate Social Communication

Mice emit ultrasonic vocalizations in different contexts throughout development and in adulthood. These vocal signals are now currently used as proxies for modeling the genetic bases of vocal communication deficits. Characterizing the vocal behavior of mouse models carrying mutations in genes associated with neuropsychiatric disorders such as autism spectrum disorders will help to understand the mechanisms leading to social communication deficits. We provide here protocols to reliably elicit ultrasonic vocalizations in pups and in adult mice. This standardization will help reduce inter-study variability due to the experimental settings. Pup isolation calls are recorded throughout development from individual pups isolated from dam and littermates. In adulthood, vocalizations are recorded during same-sex interactions (without a sexual component) by exposing socially motivated males or females to an unknown same-sex conspecific. We also provide a protocol to record vocalizations from adult males exposed to an estrus female. In this context, there is a sexual component in the interaction. These protocols are established to elicit a large amount of ultrasonic vocalizations in laboratory mice. However, we point out the important inter-individual variability in the vocal behavior of mice, which should be taken into account by recording a minimal number of individuals (at least 12 in each condition). These recordings of ultrasonic vocalizations are used to evaluate the call rate, the vocal repertoire and the acoustic structure of the calls. Data are combined with the analysis of synchronous video recordings to provide a more complete view on social communication in mice. These protocols are used to characterize the vocal communication deficits in mice lacking ProSAP1/Shank2, a gene associated with autism spectrum disorders. More ultrasonic vocalizations recordings can also be found on the mouseTube database, developed to favor the exchange of such data.

Mouse ultrasonic vocalizations are used as proxies to model the genetic bases of vocal communication deficits in mouse models for neuropsychiatric disorders. The present protocol describes three experimental contexts that reliably elicit ultrasonic vocalizations from pups (throughout development) and adult mice (same-sex interactions, male-estrus female interactions).

录制鼠标超声波发声评价社会交往

Mice emit ultrasonic vocalizations in different contexts throughout development and in adulthood. These vocal signals are now currently used as proxies ...

Analyzing Spatial Learning and Prosocial Behavior in Mice Using the Barnes Maze and Damsel-in-Distress Paradigms

The Barnes maze is a reliable measure of spatial learning and memory that does not require food restriction or exposure to extremely stressful stimuli. The Barnes maze can also assess other mouse behaviors, such as general motivation to escape from the maze platform and exploratory behavior. The Barnes maze can measure whether a genetic mutation or environmental variable can impact the acquisition and retention of spatial memories, as well as provide information about the search strategy employed by the mice. Here we use the Barnes maze to detect a memory deficit in adult mice following a single developmental ethanol exposure event. The newly described Damsel-in-Distress paradigm exposes a male mouse to a female mouse trapped in a chamber in the open center field of the arena. It provides an opportunity for the mouse to socially respond to the trapped female and exhibit prosocial behavior. The Damsel-in-Distress paradigm can also be used to examine mouse behavior in a novel arena and measure locomotor activity. Both the Barnes Maze and the Damsel-in-Distress protocols require minimal financial investment and most aspects of the tests can be constructed from common lab supplies. These flexible and accessible tools can also be used to detect behavioral changes over the course of development.

This protocol measures spatial learning and memory using the Barnes maze. A novel Damsel-in-Distress paradigm is used to assess locomotor activity and prosocial behavior in mice.

利用巴恩斯迷宫和困境中的破坏范式分析小鼠的空间学习和社会行为

The Barnes maze is a reliable measure of spatial learning and memory that does not require food restriction or exposure to extremely stressful stimuli. ...

Measuring Active and Passive Tameness Separately in Mice

Domesticated animals such as dogs and laboratory mice show a high level of tameness, which is important for humans to handle them easily. Tameness has two behavioral components: a reluctance to avoid humans (passive tameness) and a motivation to approach humans (active tameness). To quantify these components in mice, we previously developed behavioral tests for active tameness, passive tameness, and the willingness to stay on a human hand, each designed to be completed within 3 min. The data obtained were used for selective breeding, with a large number of mice analyzed per generation. The active tameness test measures the movement of the mouse toward a human hand and the contact it engages in. The passive tameness test measures the duration of time that a mouse tolerates human touch. In the stay-on-hand test, a mouse is placed on a human hand and touched slowly using the thumb of that hand; the duration of time that the animal remains on the hand is measured. Here, we describe the test set-up and apparatus, explain the procedures, and discuss the appropriate data analysis. Finally, we explain how to interpret the results.

Tameness in animals includes a reduction of avoidance responses to humans (passive tameness) and an increase in active approaches to humans (active tameness). Here, we describe detailed protocols for three behavioral tests (active tameness, passive tameness, and stay-on-hand tests) to separately measure the active and passive tameness of mice.

小鼠主动和被动驯服的测定

Domesticated animals such as dogs and laboratory mice show a high level of tameness, which is important for humans to handle them easily. Tameness has two ...

A System for Tracking the Dynamics of Social Preference Behavior in Small Rodents

Exploring the neurobiological mechanisms of social behavior requires behavioral tests that can be applied to animal models in an unbiased and observer-independent manner. Since the beginning of the millennium, the three-chamber test has been widely used as a standard paradigm to evaluate sociability (social preference) and social novelty preference in small rodents. However, this test suffers from multiple limitations, including its dependence on spatial navigation and negligence of behavioral dynamics. Presented and validated here is a novel experimental system that offers an alternative to the three-chamber test, while also solving some of its caveats. The system requires a simple and affordable experimental apparatus and publicly available open-source analysis system, which automatically measures and analyzes multiple behavioral parameters at individual and population levels. It allows detailed analysis of the behavioral dynamics of small rodents during any social discrimination test. We demonstrate the efficiency of the system in analyzing the dynamics of social behavior during the social preference and social novelty preference tests as performed by adult male mice and rats. Moreover, we validate the ability of the system to reveal modified dynamics of social behavior in rodents following manipulations such as whisker trimming. Thus, the system allows for rigorous investigation of social behavior and dynamics in small rodent models and supports more accurate comparisons between strains, conditions, and treatments.

Described here is a novel automated experimental system that offers an alternative to the three-chamber test and also solves several caveats. This system supplies multiple behavioral parameters that enable rigorous analysis of small rodent behavioral dynamics during the social preference and social novelty preference tests.

跟踪小啮齿动物社会偏好行为动态的系统

Exploring the neurobiological mechanisms of social behavior requires behavioral tests that can be applied to animal models in an unbiased and ...

<meta charset="utf8"></head><body>小鼠模型中自闭症谱系障碍的行为检测

Strategies for Assessing Autistic-Like Behaviors in Mice

Autism spectrum disorder (ASD) is a neurobiologically complex condition with a heterogeneous genetic etiology. Clinically, ASD is diagnosed by social communication impairments and restrictive or repetitive behaviors, such as hand flapping or lining up objects. These behavioral patterns can be reliably observed in mouse models with ASD-linked genetic mutations, making them highly useful tools for studying the underlying cellular and molecular mechanisms in ASD. Understanding how genetic changes affect the neurobiology and behaviors observed in ASD will facilitate the development of novel targeted therapeutic compounds to ameliorate core behavioral impairments. Our lab has employed several protocols encompassing well-described training and testing procedures that reflect a wide range of behavioral deficits related to ASD. Here, we detail two assays to study the core features of ASD in mouse models: self-grooming (a measure of repetitive behavior) and the three-chamber social interaction test (a measure of social interaction approach and preference for social novelty).

<meta charset="utf8"></head><body>评估小鼠自闭症样行为的策略

Rodent models are valuable tools for studying core behaviors related to autism spectrum disorder (ASD). In this article, we expound on two behavioral tests for modeling the core features of ASD in mice: self-grooming, which assesses repetitive behavior, and the three-chamber social interaction test, which documents social impairments.

评估小鼠自闭症样行为的策略

Autism spectrum disorder (ASD) is a neurobiologically complex condition with a heterogeneous genetic etiology. Clinically, ASD is diagnosed by social ...

使用自动图像处理方法鼠标社交性的实验评估

摘要

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跟踪小啮齿动物社会偏好行为动态的系统

评估小鼠自闭症样行为的策略

评估小鼠自闭症样行为的策略