This work was supported by the Fondation de France; by the ANR FLEXNEURIM [ANR09BLAN034003]; by the ANR [ANR- 08-MNPS-037-01-SynGen]; by Neuron-ERANET (EUHF-AUTISM); by the Fondation Orange; by the Fondation FondaMentale; by the Fondation de France; by the Fondation Bettencourt-Schueller. The research leading to this article has also received support from the Innovative Medicine Initiative Joint Undertaking under grant agreement no. 115300, resources of which are composed of financial contribution from the European Union's Seventh Framework Program (FP7/2007-2013) and EFPIA companies' in kind contribution. We thank Julie L&#233;vi-Strauss for helpful comments on the manuscript and six anonymous reviewers whose comments noticeably improved the manuscript.

伦理学声明:涉及动物主题程序已经批准科米特D&#39;Ethique恩实验安尼米尔（CETEA）N°89在巴黎巴斯德研究所。 1.动物的制备<ol><li>要录制小狗隔离来电，感兴趣的小鼠品系获得怀孕女性。注:繁殖杂男性和女性获得至少10窝，包括野生型，杂合基因敲除的幼崽获得鲁棒控制动物。 </li><li>获取两类成年小鼠的录制过程中同性相互作用发声。 <ol><li>获得至少12的男性或女性的每个基因型从感兴趣应变试验小鼠的12（考虑到个体间变异性）。注意:此协议是公适于成人的，但它可以在实验9之前进行调整，以青少年，具有降低的隔离时间。该测试适用于男性或女性。然而，要避免男性测试期从可显示在男男性社会交往测试一个明确的激进型小鼠品系。 </li><li>为了最大限度地提高affiliative相互作用的量，在实验之前隔离试验动物。房子单独男性3周（减少侵略性互动到最低限度14,17），女性为3天（E. Ey和未发表的数据），以增加他们的社会动机。 </li><li>获得男性或女性从株代表测试应变用它们作为新来者的遗传背景（ 例如，作为互动老鼠，见3.1.3）。例如，如果在C57BL / 6J背景已经生成所研究的突变体菌株，使用C57BL / 6J小鼠作为新来者。计算所需使得每个这些小鼠不使用一天的2倍以上为新来的动物的数量。房子他们组。 </li></ol></li><li>获取两类成年小鼠在发情期的女性存在来记录男性发声。 <ol><li>获取为L从感兴趣的菌株的每种基因型东12性成熟雄性（考虑到个体间变异性）。注意:如果男性从未有过的女性经验之前，把它们放在单独的笼子，让他们度过一个晚上与女性两天的测试，以增加他们的动机发出超声波发声6日前。 </li><li>获得记录的男性的背景应变性成熟女性。例如，如果正在研究已在C57BL / 6J背景产生的突变体雄性，使用C57BL / 6J雌性。计算所需使得每个这些小鼠不使用一天3次以上的女性的数目。房子他们组。 </li></ol></li></ol> 2.小​​狗隔离电话<ol><li>小狗鉴定<ol><li>出生当天预测前三天，分离出怀孕女性。 </li><li>检查出生每天早上和每天晚上的女性。注意诞生的那天为P0。 </li><li>确定使用时间持久的爪子纹身（绿色文身的皮下注射用0.3毫米×13毫米[30 G½"]针贴）在P1的幼崽。创建标志着一个，两个，三个或四个爪子代码。要为快如可能，微创扰乱幼崽，并尽快将它们放回巢越好。 </li></ol></li><li>成立了凯奇记录小狗隔离通话。 <ol><li>使用一个自制的隔音室（ 图1A）或简单的泡沫塑料盒。放置箱内的温度计来监视每个记录的温度。确保温度保持在18°C和22℃之间。 </li><li>放置一个麦克风在顶部（通过在箱的顶部的孔）。调整麦克风的高度，使麦克风的膜是12-15厘米的对话框，小狗会躺在底部上方。连接麦克风到声卡和声卡到计算机。 </li><li>要调整SOU的增益第二卡，进​​行记录试验带，不会在实验中使用的小狗。把在相同条件下的小狗如在实验（见2.3.1）。关上门。调节声音卡上的增益，使得其上的最大值（具有用于所述发声最高幅度可能），但没有超载（检查在记录软件的实时频谱图上）。 注意:如果大多数通话的幅度保持在尽可能高的水平打了几个电话可能会超载。 </li><li>注册为每个记录会话的增益级别，不幼仔/窝/会话之间改变。增益水平的变化会导致不准确的呼叫检测，并使用自动检测和测量（见5.1至5.4）的分析相同的阈值声变量的措施。 </li></ol></li></ol><img alt="图1" src="/files/ftp_upload/53871/53871fig1.jpg" /> <str翁&gt;图1:设置记录隔离从小鼠的幼崽和超声波发声的谱图调用（A）自制隔音室录制小狗的隔离调用的例子。在本呼叫类型分类所使用的不同的呼叫类型（B）的频谱图;看到说明如表1。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53871/53871fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <ol start="3"><li>进行小狗隔离的记录每两天呼叫。进行录音早晨出生在夜间幼崽，并于当天下午在白天出生的，以避免在同一个年龄组的幼崽归类与年龄的差异半天幼崽。这是最明显的非常年轻的阶段P2和P4。 <ol><li>您可以在垃圾1小狗。以最快的速度，轻轻地将其放置在用10％乙醇和干燥（直径的塑料收件人:9厘米;高度:10厘米，以防止旧的幼崽从由麦克风覆盖的区域）逃逸。把收件人刚下麦克风。 </li><li>关闭对话框迅速，默默地越好。开始在记录软件小狗超声波发声的记录（16位格式，300千赫的采样频率来捕获声音幅度高达150千赫的高品质）。 </li><li>记录的所需的时间已经过去（最多5分钟）后，停止该记录。就拿小狗开箱。写下小狗的爪子纹身。 </li><li>就拿小狗的腋温用探针温度计。马克它与带有异味少笔（水性油墨），认识到一个很小的点后面的小狗更容易在巢已记录的幼崽时，下一个选择，避免每次新的操纵所有幼崽选择。把小狗放回窝。 </li><li>清洗塑料收件人和塑料覆盖其底部有10％的乙醇，干燥得很好前把小狗旁边里面。 </li><li>选择在枯枝落叶下一个小狗，并重复2.3。 </li></ol></li><li>检查体重，运动协调，负趋地性，和发育标记（用于减少试验电池的细节，请参见在施迈瑟等人 18和Ey的等 19的方法部分）的期间1小时的休息后，以允许幼仔超声波发声的排气排放后恢复。如果完整发育试验电池如在Chadman 等人 20和Scattoni 等人 21进行使用动物的另一个队列。 </li><li>重复这些录音P2和P12之间每隔两天来表征小狗声乐的行为和发展整个头两个星期的生活。 </li></ol> 3.在同性社会交往超声波发声<ol><li>发声录音<ol><li>准备一个测试笼（50×25厘米×30℃米3;有机玻璃，100勒克斯[低强度的白光]）清洗用肥皂水，干燥和隔音室充满2厘米清新的床上用品。 <ol><li>放置使得从笼的所有角落发出叫声可以记录麦克风。固定在测试笼子的一个角落里的麦克风（无论是在笼子里或放在三脚架上），并调整角度麦克风笼底覆盖笼子的整个表面。 注:超声波探伤仪是非常方向性。 </li><li>放置在隔音室的顶部的视频摄像机来捕获测试笼子的整个表面。 注意:检查麦克风是不是隐藏在视频测试笼的一角。 </li><li>测试前，调整声卡的用备用男性和不会被在实验后所使用的备用母增益。把这些动物在记录室中的测试笼。调整声卡上的增益水平，最大限度地记录发声的幅度BU吨至减少超载所看到在录音软件实时频谱显示。 注意:增益取决于麦克风与发声动物之间的距离。 </li></ol></li><li>引进的动物进行测试（男性或女性，它会被称为"乘客"）在清新的床上用品测试笼子。离开它habituate至测试笼中的隔音室20分钟以最大限度地发挥其在3.1.3引入的未知同种的兴趣。 </li><li>这个习惯时间之后，引入交互的第二届动物（男性或女性，性别相同的乘客，但不同的耳唛/爪子纹身后辨别出来，它会被称为"新来者"）。 <ol><li>开始录制超声波发声（16位格式，300 kHz采样率捕获声音振幅高达150 kHz的高品质）以及视频捕捉介绍在测试笼子里新来的。开始录制的U习惯（单独乘员）如果需要笼勘探和社会互动中发声发射的基线电平之间的比较期间ltrasonic发声。 </li><li>通过在时间紧迫的手表（传声器近"BIP"的声音）是在什么时候新来的老鼠的后爪着地手动同步/视音频和视频录制。 </li><li>离开两种动物所需时间（例如4分钟，持续时间足以收集足够的超声波发声）相互作用。 </li></ol></li><li>把乘员和新来的人回到各自的家笼。清空使用的床上用品从测试笼，用肥皂清水冲洗，并用纸巾擦干。把新鲜的床上用品，并把它重新下一次测试隔音室。 </li></ol></li></ol> 4.男发声期间，与发情雌性互动<ol><li>测试的清晨天男性，取阴道涂片，从每一位女性来确定发情周期内的性状态。 <ol><li>尾巴抓住女性和她保持上笼网格。使用吸管以20微升的PBS冲洗阴道几次（ 即注入和回忆同20微升PBS几次）。具有相同的移液管尖回忆20微升PBS中。使用无菌PBS，以避免任何感染如果女性是连续数天进行测试。 </li><li>涂布到含有阴道细胞上的滑动悬架的PBS中。把四个样品上的一个滑板（标识个人在滑动的用铅笔的一侧）。让幻灯片在进行染色前干燥。 </li><li>实验室通风柜下工作。 <ol><li>制备纯月-Grünwald的之一浴，磷酸盐缓冲液（0.1M）和（在磷酸盐缓冲液1/20）吉姆萨一个R浴一浴。 </li><li>把载玻片在纯月-Grünwald的的浴3分钟，然后冲洗它们的1分钟的磷酸盐缓冲溶液的浴中，最后（在磷酸盐缓冲液1/20）传送它们10分钟在吉姆萨R的浴中。 </li><li>在这之后，再次冲洗载玻片在磷酸盐缓冲溶液的浴中10秒，并让它们干燥。 </li></ol></li><li>检查显微镜下的染色切片。可以在白天使用女性是那些样品目前只有大的角化上皮细胞（无细胞核，在蓝染色;全发情）。 </li></ol></li><li>测试之前把男性在试验室至少30分钟。 </li><li>发声录音<ol><li>如有必要，重复3.1。 </li><li>介绍要测试的男性（新鲜床上用品）。离开他habituate至测试笼中的隔音室10分钟。 </li><li>这个习惯时间之后，引入在发情期女性（从染色选择那些中）。 <ol><li>开始录制超声波发声和视频捕捉到在测试笼子里引进的女性。开始习惯（雄性单独）如果需要笼勘探和社会互动中发声发射的基线电平之间的比较期间记录所述超声波发声。 </li><li>通过在时间紧迫的手表（传声器近"BIP"的声音）恰好在雌性小鼠的后爪着地手动同步/视音频和视频录制。 </li><li>离开两种动物所需时间（例如4分钟，持续时间足以收集足够的超声波发声）相互作用。 </li></ol></li><li>把阳和阴背部在其各自的家笼中。清空使用的床上用品从测试笼，用肥皂清水冲洗，并用纸巾擦干。把新鲜的床上用品，并把它重新下一次测试隔音室。 注:这是最佳的每一天只能一次使用每个发情雌性（但如果需要它可以使用高达3倍的同一天，但不是在一个行）。 </li></ol></li></ol> 5.变量要提取<ol><li>准备音频文件的分析。注意:下面的步骤是特定的一种visoft SASLab临 ，并且可以根据所使用的软件变化。 <ol><li>切文件，以便它们在时间腕表"BIP"恰好开始，并且所需的持续时间（5分钟的小狗录像，录音成人4分钟）之后结束。 </li><li> （;高通以30 kHz的频率切断编辑&gt;滤镜&gt; FIR时域滤波器）通过使用高通滤波器滤除在30千赫的幅度。使用批处理来筛选的利息（操作&gt;批处理&gt; FIR滤波器）的所有文件。 </li><li>通过与软件被贴上标识每个超声波发声。 <ol><li>使用小狗录音自动检测（工具&gt;标签&gt;创建波形事件部分标签）。调整门槛，保持时间和边缘f还是最准确的检测。手动检查检测，必要时（推荐）调​​整标签。 </li><li>使用成人用录音的背景噪音的视觉检测（标签手动插入）​​（选择发声，点击右键，然后从标记插入分节标签）。 </li></ol></li><li>创建谱图。激活自动参数测量（工具&gt;自动参数测量&gt;自动参数测量设置）。 </li><li>选中"启用自动测量"，"从整个频谱计算参数"和"自动更新"复选框。选择"元素分离":以交互方式（部分标签）。 </li><li>复选框来计算所需的时间参数，并基于频谱的参数（峰值频率）（元，时间间隔，开始/结束时间的持续时间），以及测量位置（元素开始，元素结束，平均值，最大值，最小值） 。 </li><li>复制测量并粘贴米的电子表格。 注意:对于与成人进行录音，基于频谱参数测量可能会因为背景噪音是不可能的。通过点击不同的频率值上直接频谱和在表手动粘贴的值，用峰值频率的手动测量。 </li></ol></li><li>通过首先确定发射发声的数量（标签总数）确定呼叫速率， 即，每分钟的呼叫数。然后，除以该文件的持续时间（以分钟为单位）记录的发声总数计算出的传输率。 </li><li>确定时间的组织， 即调用来确定序列组织之间的时间间隔分布。 <ol><li>计算发声n的端部和发声的n + 1的使用各标签的开始/结束时间的开始之间的时间间隔。 </li><li>建立了Ultrason之间的时间间隔的分布密度IC发声。 </li></ol></li><li>确定通话剧目， 即定义出现在记录的呼叫类型。使用分级在表1和图1B介绍的例子。 <ol><li>当5.1.3标记各个发声，写在标签中的呼叫类型的名称。 </li><li>计算出准确的数量和每个呼叫类型建立声剧目的比例。 </li></ol></li><li>确定每个发声， 即，持续时间，峰值频率的声学特性（ 例如，具有最高振幅频率）在开始和呼叫，最大和最小峰值频率的端部，并平均频率，如果自动测量是可能的（ 图1B） 。 <ol><li>使用自动参数测量功能，在软件中自动测量持续时间，峰值频率特性小狗的记录（ 例如 ，开始，结束，平均值，最大值，最小值）。 </li&gt; <li>使用的标签的持续时间作为发声成人记录的持续时间。手动测量谱图窗口中的峰值频率特性（ 例如，开始，结束，最大值，最小值）上。 </li></ol></li><li>夫妇超声波发声数据和社交互动数据（从冰冷的平台22 MiceProfiler插件）。 <ol><li>确保为精确同步地将音频和在协议中所描述的视频记录。 </li><li>使用ICY平台的小鼠探查器跟踪插件在肖蒙等 22所描述的编码社会互动的视频。恰好开始跟踪时引入的动物的后爪接触地面。 </li><li>上传编码的视频文件和相应的XML文件中的ICY平台的小鼠探查视频标签制作插件（通过小鼠探查器跟踪生成）在肖蒙等描述人22。 注: 老鼠探查视频标签制作插件会自动链接视频文件和音频文件的分析产生的，如果他们有相同名称的文本文件（参见:http://icy.bioimageanalysis.org/plugin/Mice_Profiler_Video_Label_Maker） 。 </li><li>检查鼠标规模是正确后，点击"创建USV统计"为每个文件，以获得数量和分离的文件中的每个社交活动时发出发声的比例。 </li></ol></li></ol> 6.上传文件的mouseTube数据库<ol><li>确保文件是可以从机构外部访问存储服务器上。 注:在一些机构高安全级别的托管的服务器需要一个特定的配置，以从机构以外人们连接访问。 </li><li>进入mouseTube网站（http://mousetube.pasteur.fr）。登录（登录本第二密码由管理员分配给每个用户）。 </li><li>检查已经记录的小鼠品系是否通过点击"应变"按钮，在mouseTube数据库中存在。如果没有，请管理员来创建它。 </li><li>创建使用科目"科目&gt;创建"按钮。输入记录动物的识别码。收集他们组来缓解数据进行检索。 </li><li>输入用于记录使用超声波发声"协议&gt;创建"按钮协议的描述。 </li><li>使用每个记​​录会话创建实验"实验&gt;创建"按钮。指定协议，这已被记录，硬件和使用的软件和其特异性的组的个人。 注:实验收集所有相应的发声文件的元数据。 </li><li>创建使用"Vocalisations链接到发声文件&gt;新建＆＃34;按钮。选择列表中的实验。复制并粘贴发声文件的URL（此链接以http:// ...）插入的相应字段"文件链接"一栏。通过点击按钮，验证项目"创建mouseTube和文件之间的链接。" 注意:它不要求在同一时间，以填补对每个文件中的每个框。 </li><li>如有必要，修改在任何时候进入的链接，而"注意事项"一节中添加详细信息。不要犹豫，写下笔记透露更多的细节。举例来说，如果朝这与音频文件同时记录的视频文件的链接已输入，这可以在"说明"中指定。 </li></ol>

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The authors have nothing to disclose.

这里介绍的协议提供了标准化的，可靠的方式来收集老鼠的超声波发声实验室。这些非常有限的情况下，目前标准化的优势。它们被成功用于比较株18,19,26,27内菌株或基因型。正如代表提出的结果，这些方法让非典型社会沟通的突变为Shank2，与自闭症有关的基因小鼠的鉴定。小鼠品系，不同的上下文实验室之间或者甚至之间的比较将被mouseTube数据库在较大的数据集的可用性被触发。这个工具应该允许多元分析提高鼠标的超声波发声的研究。 这里描述的方案进行了优化，一个应变内测试不同基因型的小鼠，因为它是在大多数研究完成建模遗传Çontribution为神经精神障碍。建议设计实验研究每一个有可能的最好的控制。事实上，垃圾影响可能掩盖或人为夸大遗传效应28,29。因此，建议包括同窝控制每个基因型。因此育种杂家长应该受到青睐，因为它将使突变体和对照组小鼠的窝内的正确匹配。这证明了爪子纹身所有幼崽的标记（盲基因型）在整个录音跟踪个人每两天。基因分型在断奶完成，通过取尾样品。当从P2上录制小狗隔离来电，我们不建议服用尾样本已经幼崽，因为该操作包括补充操纵和压力非常接近的时间来记录会话。 这里建议的协议引发超声波发声成人不允许的EMI明确确定在发声的垃圾。这就解释了为什么我们操纵试验动物的动机。事实上，测试小鼠中分离，而不是新来的和试验动物habituate期间同性相互作用时间长到测试笼中。在男女互动中，介绍了女性不是孤立的，测试的男性habituates更短的时间，因为动机可能是在这种情况下性更高。动机的这些操作应尽可能测试鼠标发射的发声和不引入的一个的概率。要记录在性方面男性超声波发声，与发情雌性新鲜的（ 即不冻结）尿一个简单的棉签也可以在笼子里30推出。此方法允许超声发声到测试男性用100％的确定性的分配，但它可以防止收集关于这些发声的发射的实际社会背景的任何具体信息。因此，我们赞成protoco升这里描述（具有可自由移动的发情女性）。我们还建议从突变株的小鼠进行测试时，始终使用一脉相承引入小鼠和分析数据，一对老鼠发声的。最近的一项研究推广使用三角测量来定位发射器31。在这项研究中，发现女性与男性接触时也发出超声波发声。这可能是由以下事实，他们在记录会话之前分离为至少两个星期来解释。使用在本研究中提出的三角测量的泛化仍应允许在大多数情况下，发声的发射极的识别如果录像正确同步。 从开发过程中记录的幼仔隔离呼叫不从被褥背景噪音的干扰。通常一个自动分析作品非常好，提取的主要变量。与此相反，从大人记录发声是d-从动物的被褥移动背景噪声isturbed。自动分析可能会失败，因此应采用人工分析。然而，在测试笼子加入被褥应当提供用于动物比裸土压力较小（老鼠不喜欢）的条件。在社区进一步努力都集中在改善超声波发声的自动检测在各种条件下，即使是那些暗示背景噪声。例如，语音软件允许分析已被手动选择了没有背景噪声32的发声。在该软件中，声变量的提取是自动但需要的手动初始选择。 应当指出的是，个体间变异性是在小鼠的发声特性非常重要。例如，在一个发情女性的存在成年男性的呼叫率是非常分布（ 图1）。我们苏ggest这些标准化协议已经引发超声波发声来限制有关的实验内容的可变性。尽管如此，我们想指出的提出不仅均值和SEM的数据的重要性，但最重要的是体积小33样品的各个点。这也是非常相关 - 如果不是必要 - 记录每个组/基因型的至少12个人收集的代表性数据。在许多情况下，个体间变异不应被隐藏（通常它不可能是），它可能是非常重要，以确定携带所研究的基因突变，但不显示任何非典型表型的个体。这样的个人可以提供有关补偿，这可能会打开新的通路遗传性疾病的治疗线索。 在小鼠模型的神经精神障碍，声乐行为和社会交往的行为大部分是刻画考虑ED外（ 如 19,27,34,35）。现在最近的分析方法的相互作用（使用MiceProfiler例如）36，以及这种分析与来自录音数据结合的可能性期间提供的社交活动和事件的序列的半自动详细的表征。该方法的主要优点是提供了社会的通信的综合视图中的ASD的小鼠模型，以更精确地确定哪些社会交往的各方面都受到影响。在本协议的同步是仍然手册，但此可通过音频记录软件触发视频记录的提高。这种类型的分析应该成为提供在神经精神障碍的小鼠模型中的社会交流障碍的更全面的标准。此外，到现在为止，歌声信号大多从发射极侧进行分析（ 即 ，测试都建有利于沃的发射被测试鼠标校准信号，如在本协议）。重点应现在也对这些信号的接收器进行设置，以更好地识别这些声信号的功能。这应该由也评价在本协议的新来的小鼠在成人的行为（用MiceProfiler例如）36，通过使用播放实验16来完成，或通过建立新的协议。实际上，本协议提供可能不反映在小鼠发声排放的确切行为学条件非常受限制的情况。超声波发声的自发辐射将不得不使用连续录音录像摆脱对小鼠的自发行为声乐更多的光线较好的特点。

患者神经精神障碍通常显示社会交往障碍（ 如 ，患有自闭症，精神分裂症或阿尔茨海默病）1。遗传工程小鼠越来越经常用于这些病症2的遗传原因建模。在这些小鼠模型研究社会的沟通是高利息的了解基因突变导致的非典型社会功能障碍的机制和测试新的疗法。因为老鼠是群居动物，并用嗅觉，触觉，视觉和听觉信号中的每个人沟通，他们是合适的模型来评估社会沟通。 现在鼠标超声波发声目前用于作为代理为有声通信赤字3,4-（但发声学习在这种仍有争议5,6-存在的遗传基础建模，即使大多数最近的研究的申辩E对于没有声乐学习七 ）。实验室老鼠被发现发射出超声波发声的母婴关系中，男性与女性的社会性互动，同性社交互动（参考点评 ）和少年，少年的社会交往9。鼠标幼仔从大坝和同窝10孤立时，他们的头两个星期的生活中散发出的隔离通话。男性发射出超声波发声的（从她或泌尿线索）11,12的发情女性在场的时候。男性和女性与同性13,14的同种未知交互时发射出超声波发声。该组织与这些发声的功能尚不完全清楚，需要进一步调查。在功能方面目前的知识仅限于检索行为的启发在母亲听到小狗隔离电话，对成年男性vocalizat成年女性接近的便利离子15和成年男性听力成年女性发声16增加的探索行为。 表征在神经精神障碍的小鼠模型有声通信的异常应在标准条件下进行，以排除的实验条件重大贡献。这样的表征，同时与社会交往和神经生物学研究，在各种遗传模型的评价相结合应该提高我们对鼠标的超声波通信的不同方面的遗传贡献知识。在长期，它应该对社会交往的人类神经生物学的一些基地给予进一步的光。目前，我们的目标是提供简单的协议，在开发过程中，并在成年后在实验室雄性和雌性小鼠可靠引起超声波发声。这样的协议将缓解录音更可靠地比较ULTR标准化株和实验室之间asonic发声排放。还应该促进这种录音中具有鼠标超声波发声录音没有现成的经验实验室设立。我们还突出显示当前可能性超声波发声数据与在成年小鼠社会互动同时收集，以获得对社会障碍以及对超声波发声的排放的情况下的关键信息的详细行为数据相结合。这种分析会的组织和老鼠的超声波发声的功能，新的视角。最后，我们还发布给mouseTube数据库（http://mousetube.pasteur.fr）与整个科学界分享超声波发声录音的可能性。音频记录数据开放访问应该允许科学家记录在其他拉波超声波发声比较自己的数据提升鼠标超声波通信知识ratories（具有类似的或不同的菌株/协议）和/或挑战他们的分析方法不同的条件下记录的文件。

<table><tbody><tr><td>needles 0.3mm x 13 mm [30&amp;nbsp;G 1/2&quot;]</td><td>BD Microlance</td><td>304000</td><td>-</td></tr><tr><td>green tattoo paste</td><td>Ketchum Manufacturing Inc., Ottawa, Canada</td><td>329AA</td><td>-</td></tr><tr><td>thermometer</td><td>Fisherbrand, Waltham, USA</td><td>4126 (W255NA)</td><td>-</td></tr><tr><td>self-made soundproof chamber (pups)</td><td>Institut Pasteur, Paris</td><td>-</td><td>acoustic foam + plexiglas; inside dimensions (W x H x D): 32 cm x 33 cm x 32 cm</td></tr><tr><td>small surface thermister + single probe thermocouple</td><td>Harvard Apparatus</td><td>599814 + 601956</td><td>-</td></tr><tr><td>smell-less pen</td><td>for instance: Giotto</td><td>-</td><td>ink made with water, washable: these pens are designed for babies</td></tr><tr><td>Ethanol absolute (100%)</td><td>Sigma Aldrich,&amp;nbsp; Saint-Quentin Fallavier, France</td><td>24103</td><td>diluted 1/10</td></tr><tr><td>Condenser ultrasound microphone Avisoft-Bioacoustics CM16/CMPA</td><td>Avisoft Bioacoustics, Berlin, Germany</td><td>#40011</td><td>furnished with extension cables by the Avisoft company</td></tr><tr><td>Ultrasound Gate 416H</td><td>Avisoft Bioacoustics, Berlin, Germany</td><td>#34163</td><td>sound card</td></tr><tr><td>Avisoft Recorder USGH</td><td>Avisoft Bioacoustics, Berlin, Germany</td><td>#10301; #10302</td><td>recording software for Windows Vista, 7 and 8</td></tr><tr><td>Avisoft SASLab Pro</td><td>Avisoft Bioacoustics, Berlin, Germany</td><td>#10101, 10111; #10102, 10112;&amp;nbsp;</td><td>Windows 10, 8.1, 8, 7 or Vista including Intel-based Apple Macintosh running Boot Camp, Parallels or similar virtualization software.</td></tr><tr><td>Laptop or Apple Macintosh running Boot Camp</td><td>-</td><td>-</td><td>running Windows 10, 8.1, 8, 7 or Vista; for the Apple Macintosh, Boot Camp is preferred to virtualizations softwares such as Parallels due to memory constraints</td></tr><tr><td>plastic recipient (pup recordings)</td><td>Lock &amp;amp; Lock, Chatswood, USA</td><td>HPL932D</td><td>Lock &amp;amp; Lock Stackable Airtight Container Round 700&amp;nbsp;ml; use without the cover; dimensions: 9&amp;nbsp;cm diameter, 10 cm height</td></tr><tr><td>PBS 1x (pH = 7.4)</td><td>Gibco (Life Technologies)</td><td>10010-023</td><td>-</td></tr><tr><td>slides</td><td>Menzel-Gl&amp;auml;ser, Thermo Scientific</td><td>J1800AMNZ</td><td>Superfrost Plus</td></tr><tr><td>May-Gr&amp;uuml;nwald solution 500 ml</td><td>RAL R&amp;eacute;actifs, Martillac, France</td><td>320070-0500</td><td>-</td></tr><tr><td>Giemsa R 500 ml</td><td>RAL R&amp;eacute;actifs, Martillac, France</td><td>720-1107</td><td>diluted 1/20 in phosphate buffer solution</td></tr><tr><td>phosphate buffer solution (self-made)</td><td>-</td><td>-</td><td>pH = 7, 0.1 M: 39 ml NaH&lt;sub&gt;2&lt;/sub&gt;PO&lt;sub&gt;4&lt;/sub&gt; 0.2 M + 61 ml Na&lt;sub&gt;2&lt;/sub&gt;HPO&lt;sub&gt;4&lt;/sub&gt; 0.2 M + 100&amp;nbsp;ml H&lt;sub&gt;2&lt;/sub&gt;O (final volume: 200&amp;nbsp;ml)</td></tr><tr><td>test cage</td><td>Institut Pasteur, Paris</td><td>-</td><td>50&amp;nbsp;x&amp;nbsp;25&amp;nbsp;cm, 30&amp;nbsp;cm height; Plexiglas</td></tr><tr><td>self-made soundproof chamber (adult recordings)</td><td>Institut Pasteur, Paris</td><td>-</td><td>acoustic foam + PVC; inside dimensions (W x H x D): 66 cm x 90 cm x 46 cm</td></tr><tr><td>video camera</td><td>From Noldus Information Technologies, Wageningen, The Netherlands</td><td>-</td><td>high-resolution CamTech Super-Hi-Res video camera; 25 fps&amp;nbsp;</td></tr><tr><td>EthoVision XT</td><td>Noldus Information Technology, Wageningen, The Netherlands</td><td>http://www.noldus.com/animal-behavior-research/products/ethovision-xt</td><td>video acquisition software</td></tr><tr><td>Mice Profiler Tracker plugin from the ICY platform</td><td>Bio Image Analysis, Institut Pasteur, Paris</td><td>http://icy.bioimageanalysis.org/plugin/Mice_Profiler_Tracker</td><td>tracking software to analyse behavioral events during social interactions</td></tr></tbody></table>

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.

本协议，我们的特点缺乏ProSAP1 / Shank2，自闭症谱系障碍（ASD）23-25 ​​相关的基因小鼠的声乐行为。 ASD是由在社会交往赤字和刻板行为特点1。我们Shank2 - / -小鼠显示多动，焦虑加重和非典型声乐通信18,26。事实上，我们注意到，Shank2 - / -小鼠与它们的野生型同窝典型倒U形曲线的比较显示的小狗隔离呼叫他们的发射率非典型发育轮廓Shank2 - / -小鼠显示增加的呼叫速率在P4和拆借利率P6与它们的野生型同窝（ 图2）相比下降。我们也观察到女性的相互作用降低拆借利率涉及Shank2 - / -女性在试比较Ñ ​​与涉及野生型同窝（ 图2）的相互作用。我们研究了5种不同的呼叫类型的剧目。它似乎是幼仔之间不同（例如这里P2，P6和P10）和成人（ 图3）。基因型相关差异显著主要是在成年期。在涉及成年Shank2社交互动- / -男性或女性用的C57BL / 6N雌性，更短的呼叫和非结构化呼叫记录了与涉及它们的野生型同窝小鼠（ 图3D和E）的相互作用比较。较不复杂的呼叫和跳频的呼叫也被与涉及成年Shank2一个C57BL / 6N雌性相互作用期间记录- / -雌性相比与涉及Shank2 + / +雌性（ 图3E）的相互作用。最后，我们还测定了手动声学变量。有没有显著的基因型相关差异杜圈发展。与此相反，涉及成年Shank2相互作用期间记录的呼叫的持续时间- / -雌性比那些涉及它们的野生型同窝小鼠（ 图4A）的相互作用过程中记录短。我们还强调，超声波发声的峰值频率不显著的基因型相关差异26小狗在开发过程中增加。在涉及Shank2相互作用- / -男性或女性用的C57BL / 6N雌性，超声波发声与涉及它们的野生型同窝小鼠（ 图4B）的相互作用过程中记录的呼叫相比具有较低的峰值频率。 此外，本方案还允许通过从录音的数据组合以从MiceProfiler（ICY软件，研究所过去提取的行为数据来研究超声波发声的排放的情况下欧元，巴黎）。例如，在女性雌相互作用，最超声波发声被发射时动物是在接触，更具体乘员嗅探新来的肛门生殖器区域，或至少乘员作为新来的后面。小鼠也发出许多超声波发声当乘客走近新来的（ 图5，上图）。 / - -小鼠与新来的身体接触（ 例如 ，嗅着新来的肛门生殖器区域），比当乘客是野生型小鼠在乘员Shank2少发声记录。 / - -鼠标时相比，这是一个野生型小鼠时，新来背后的乘员是一个Shank2少发声被触发。更发声也记录，当新来的人是在乘员小鼠的视场，更多的是在野生型相比在突变体（ 图5，下图）。 S ="jove_content"FO: - together.within页保留="1"&gt; <img alt="图2" src="/files/ftp_upload/53871/53871fig2.jpg" /> 图 2:在开发过程中和在成年男性和女性 Shank2 超声波发声的发射率 - / - 小鼠和野生型同窝呼叫幼仔的速率（每两天从P2到P12中，n = 18-19 Shank2 + / +， N = 15-16 Shank2 - / -在男性发情母相互作用）和成年人（N = 15 Shank2 + / +，N = 16 Shank2 - / - ）和女女的相互作用（N = 15 Shank2 + / +，N = 13 Shank2 - / - ）中的野生型小鼠（左面板）和Shank2 - / -小鼠（右图）。数据表示为平均值+/- SEM和个别点（非成对的Wilcoxon测试:* P &lt;0.05，** P &lt;0.01，*** P &lt;0.001）。ge.jpg"目标="_空白"&gt;点击此处查看该图的放大版本。 <img alt="图3" src="/files/ftp_upload/53871/53871fig3.jpg" /> 图 3:Shank2声乐曲目 - / -由P2幼崽发出的五种不同的呼叫类型小鼠和野生型同窝比例（A，N = 20 Shank2 + / +，N = 18 Shank2 - / - ），P6幼崽（B; N = 19 Shank2 + / +，N = 18 Shank2 - / - ），P10幼仔（C; N = 20 Shank2 + / +，N = 18 Shank2 - / - ），成年男性具有发情雌性（D ; N = 16 Shank2 + / +，N = 16 Shank2 - / - ）和成年女性与另一雌（E; N = 15 Shank2 + / +，N = 13 Shank2 </eM&gt; -野生型小鼠（左小图）和Shank2） - - / / -小鼠（右图）。数据显示为平均值+/- SEM和个别点（卡方检验:* P &lt;0.05，** P &lt;0.01，*** P &lt;0.001）。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53871/53871fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/53871/53871fig4.jpg" /> 图 4:从超声波发声的 Shank2 提取声变量 - / - 小鼠和野生型同窝 （A）中的所有呼叫类型的时间混淆由P2幼崽发出的（N = 20 Shank2 + / +，N = 18 Shank2 - / - ），P6幼仔（N = 19 Shank2 + / +，N = 18 Shank2 - / - ），P10幼仔（N = 20 Shank2 + / + </sUP&gt;，N = 18 Shank2 - / - ），成年男性与发情雌性（N = 16 Shank2 + / +，N = 16 Shank2 - / - ）和成年女性与其他女性（N = 15 Shank2 + / +， N = 13 Shank2 - / - ）中的野生型小鼠（左面板）和Shank2 - / -小鼠（右图）。所有的呼叫类型测量（B）的最大峰值频率P2幼仔，幼仔P6，P10幼仔，成年男性困惑与发情雌性和成年女性与另一名女性（相同Ns个如上）。数据显示为平均值+/- SEM和个别点（非成对的Wilcoxon测试:* P &lt;0.05，** P &lt;0.01，*** P &lt;0.001）。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53871/53871fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图5" src="/files/ftp_upload/53871/53871fig5.jpg" /> 图5:上下文通过涉及对发出的超声波发声的女性，成年女性的社会交往鼠标超声波发声发射第比例Shank2 + / +与C57BL / 6N小鼠（N = 16，A），并涉及Shank2双- / -与一个C57BL / 6N小鼠（N = 13，b）在以下类型的行为事件（红:乘员，绿色环保:新来的）:社会交往，口-口接触，从乘员鼠肛门生殖器嗅探，ano-生殖器由新来的鼠标嗅探，后面新来的，后面的乘员新来的，乘客不动，新来的不动，方法与乘员及乘客来自新来逃避，方法与新来的＆来自乘客，方法逃避与来自乘客，方法逃避与来自新来逃生，乘客按照新来的，新来的乘员，乘员的视野中新来的视野。数据是p不满为平均值+/- SEM和个别点（非成对的Wilcoxon测试:* P &lt;0.05，** P &lt;0.01）。未公布的数据， <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53871/53871fig5large.jpg" target="_blank">请点击这里查看该图的放大版本。</a> <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td> 呼叫类型 </td><td> 描述 </td></tr><tr><td>短</td><td>持续时间≤5毫秒和频率范围≤6.25千赫</td></tr><tr><td>简单</td><td>持续时间&gt; 5毫秒和频率范围≤6.25千赫（平），或频率调制仅在一个频率范围&gt; 6.25 kHz的方向（向上或向下） </td></tr><tr><td>复杂</td><td>在多于一个方向上和频率范围&gt; 6.25千赫频率调制（调制），或包括一个或多​​个附加频率的component（谐波或非线性现象，但没有饱和），但对频率范围无约束（配合物） </td></tr><tr><td>频率跳跃</td><td>包括一次跳跃（一个频率跳变）以上的跳跃（频率跳跃，其他），频率不连续的频率分量，与之间的时间间隔（混合）或无纯音呼叫内的任何部分吵</td></tr><tr><td>非结构化</td><td>没有纯音成分标识; "噪音"电话</td></tr></tbody></table> 表1:五种类型的老鼠的超声波发声的特性用于确定鼠标超声波发声内5个不同的呼叫类型持续时间，频率范围，频率调制和频率跳跃的标准范例。

Watch this Scientific Journal Video about Recording Mouse Ultrasonic Vocalizations to Evaluate Social Communication at JoVE.com

Recording Mouse Ultrasonic Vocalizations to Evaluate Social Communication

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 ...

recording-mouse-ultrasonic-vocalizations-to-evaluate-social

Universidad de Cartagena UDEC

Research

JoVE Journal

Behavior

22.2K 次观看.  University Paris Diderot, CNRS UMR 3571, Institut Pasteur. 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).

视频: Recording Mouse Ultrasonic Vocalizations to Evaluate Social Communication

Using Chronic Social Stress to Model Postpartum Depression in Lactating Rodents

Exposure to chronic stress is a reliable predictor of depressive disorders, and social stress is a common ethologically relevant stressor in both animals and humans. However, many animal models of depression were developed in males and are not applicable or effective in studies of postpartum females. Recent studies have reported significant effects of chronic social stress during lactation, an ethologically relevant and effective stressor, on maternal behavior, growth, and behavioral neuroendocrinology. This manuscript will describe this chronic social stress paradigm using repeated exposure of a lactating dam to a novel male intruder, and the assessment of the behavioral, physiological, and neuroendocrine effects of this model. Chronic social stress (CSS) is a valuable model for studying the effects of stress on the behavior and physiology of the dam as well as her offspring and future generations. The exposure of pups to CSS can also be used as an early life stress that has long term effects on behavior, physiology, and neuroendocrinology.

This article describes the use of chronic resident intruder social stress as an ethologically relevant paradigm to model postpartum depression and anxiety in lactating rodents.

哺乳期鼠害使用慢性社会压力型号产后抑郁症

Exposure to chronic stress is a reliable predictor of depressive disorders, and social stress is a common ethologically relevant stressor in both animals ...

科研

行为学

Assessment of Ultrasonic Vocalizations During Drug Self-administration in Rats

Drug self-administration procedures are commonly used to study behavioral and neurochemical changes associated with human drug abuse, addiction and relapse. Various types of behavioral activity are commonly utilized as measures of drug motivation in animals. However, a crucial component of drug abuse relapse in abstinent cocaine users is "drug craving", which is difficult to model in animals, as it often occurs in the absence of overt behaviors. Yet, it is possible that a class of ultrasonic vocalizations (USVs) in rats may be a useful marker for affective responses to drug administration, drug anticipation and even drug craving. Rats vocalize in ultrasonic frequencies that serve as a communicatory function and express subjective emotional states. Several studies have shown that different call frequency ranges are associated with negative and positive emotional states. For instance, high frequency calls ("50-kHz") are associated with positive affect, whereas low frequency calls ("22-kHz") represent a negative emotional state. This article describes a procedure to assess rat USVs associated with daily cocaine self-administration. For this procedure, we utilized standard single-lever operant chambers housed within sound-attenuating boxes for cocaine self-administration sessions and utilized ultrasonic microphones, multi-channel recording hardware and specialized software programs to detect and analyze USVs. USVs measurements reflect emotionality of rats before, during and after drug availability and can be correlated with commonly assessed drug self-administration behavioral data such lever responses, inter-response intervals and locomotor activity. Since USVs can be assessed during intervals prior to drug availability (e.g., anticipatory USVs) and during drug extinction trials, changes in affect associated with drug anticipation and drug abstinence can also be determined. In addition, determining USV changes over the course of short- and long-term drug exposure can provide a more detailed interpretation of drug exposure effects on affective functioning.

Drug self-administration and ultrasonic vocalizations (USV) are used as behavioral assessments in animal research, but rarely in combination. The purpose of this article is to describe the advantages of recording USVs during drug self-administration procedures to assess affective responses to drug experience.

在大鼠的药物自我管理的评估超声发声

Drug self-administration procedures are commonly used to study behavioral and neurochemical changes associated with human drug abuse, addiction and ...

神经科学

Experimental Assessment of Mouse Sociability Using an Automated Image Processing Approach

Mouse is the preferred model organism for testing drugs designed to increase sociability. We present a method to quantify mouse sociability in which the test mouse is placed in a standardized apparatus and relevant behaviors are assessed in three different sessions (called session I, II, and III).
The apparatus has three compartments (see Figure 1), the left and right compartments contain an inverted cup which can house a mouse (called &#34;stimulus mouse&#34;).
In session I, the test mouse is placed in the cage and its mobility is characterized by the number of transitions made between compartments. In session II, a stimulus mouse is placed under one of the inverted cups and the sociability of the test mouse is quantified by the amounts of time it spends near the cup containing the enclosed stimulus mouse vs. the empty inverted cup. In session III, the inverted cups are removed and both mice interact freely. The sociability of the test mouse in session III is quantified by the number of social approaches it makes toward the stimulus mouse and by the number of times it avoids a social approach by the stimulus mouse.
The automated evaluation of the movie detects the nose of the test mouse, which allows the determination of all described sociability measures in session I and II (in session III, approaches are identified automatically but classified manually). To find the nose, the image of an empty cage is digitally subtracted from each frame of the movie and the resulting image is binarized to identify the mouse pixels. The mouse tail is automatically removed and the two most distant points of the remaining mouse are determined; these are close to nose and base of tail. By analyzing the motion of the mouse and using continuity arguments, the nose is identified.
<img alt="Figure 1" src="/files/ftp_upload/52508/52508fig1.jpg" /> 
Figure 1. Assessment of Sociability During 3 sessions. Session I (top): Acclimation of test mouse to the cage. Session II (middle): Test mouse moving freely in the cage while the stimulus mouse is enclosed in an inverted cup. Session III (bottom): Both test mouse and stimulus mouse are allowed to move freely and interact with each other.

This protocol describes a method to quantify mouse sociability. Mice are videotaped as they move and interact in a special cage. Movie processing allows for the automated quantification of sociability with excellent accuracy and reliability.

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 ...

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 ...

Determining Ultrasonic Vocalization Preferences in Mice using a Two-choice Playback Test

Mice emit ultrasonic vocalizations (USVs) during a variety of conditions, such as pup isolation and adult social interactions. These USVs differ with age, sex, condition, and genetic background of the emitting animal. Although many studies have characterized these differences, whether receiver mice can discriminate among objectively different USVs and show preferences for particular sound traits remains to be elucidated. To determine whether mice can discriminate between different characteristics of USVs, a playback experiment was developed recently, in which preference responses of mice to two different USVs could be evaluated in the form of a place preference.
First, USVs from mice were recorded. Then, the recorded USVs were edited, trimmed accordingly, and exported as stereophonic sound files. Next, the USV amplitudes generated by the two ultrasound emitters used in the experiment were adjusted to the same sound pressure level. Nanocrystalline silicon thermo-acoustic emitters were used to play the USVs back. Finally, to investigate the preference of subject mice to selected USVs, pairs of two differing USV signals were played back simultaneously in a two-choice test box. By repeatedly entering a defined zone near an ultrasound emitter and searching the wire mesh in front of the emitter, the mouse reveals its preference for one sound over another. This model allows comparing the attractiveness of the various features of mouse USVs, in various contexts.

Ultrasonic vocalizations (USVs) in mice differ depending on age, sex, condition, and genetic background. Using two ultrasound emitters broadcasting simultaneously in different locations, this two-choice test can evaluate murine recognition and preference responses to different characteristics of USVs.

采用双选择播放测试确定超声波发声首小鼠

Mice emit ultrasonic vocalizations (USVs) during a variety of conditions, such as pup isolation and adult social interactions. These USVs differ with age, ...

The Social Dimension of Stress: Experimental Manipulations of Social Support and Social Identity in the Trier Social Stress Test

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations supportive behavior of other people as well as positive social relationships can act as powerful resources to cope with stress. In order to study the interplay between these variables, this protocol describes two effective experimental manipulations of social relationships and supportive behavior in the laboratory. In the present article, these two manipulations are implemented in the Trier Social Stress Test (TSST)&#8212;a standard stress induction paradigm in which participants are subjected to a simulated job interview. More precisely, we propose (a) a manipulation of the relationship between different protagonists in the TSST by making a shared social identity salient and (b) a manipulation of the behavior of the TSST-selection committee, which acts either supportively or unsupportively. These two experimental manipulations are designed in a modular fashion and can be applied independently of each other but can also be combined. Moreover, these two manipulations can also be integrated into other stress protocols and into other standardized social interactions such as trust games, negotiation tasks, or other group tasks.

Previous research on the social dimension of stress has focused on two important variables: social identity and social support. This protocol introduces an effective experimental manipulation of these two social variables and describes their implementation in a standard stress induction paradigm (Trier Social Stress Test).

应力的社会问题:社会支持和社会认同的特里尔社会的实验操作压力测试

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations ...

Experimental Research Examining How People Can Cope with Uncertainty Through Soft Haptic Sensations

Human beings are constantly surrounded by uncertainty and change. The question arises how people cope with such uncertainty. To date, most research has focused on the cognitive strategies people adopt to deal with uncertainty. However, especially when uncertainty is due to unpredictable societal events (e.g., economical crises, political revolutions, terrorism threats) of which one is unable to judge the impact on one&#8217;s future live, cognitive strategies (like seeking additional information) is likely to fail to combat uncertainty. Instead, the current paper discusses a method demonstrating that people might deal with uncertainty experientially through soft haptic sensations. More specifically, because touching something soft creates a feeling of comfort and security, people prefer objects with softer as compared to harder properties when feeling uncertain. Seeking for softness is a highly efficient and effective tool to deal with uncertainty as our hands are available at all times. This protocol describes a set of methods demonstrating 1) how environmental (un)certainty can be situationally activated with an experiential priming procedure, 2) that the quality of the softness experience (what type of softness and how it is experienced) matters and 3) how uncertainty can be reduced using different methods.

To date research has focused on cognitive strategies people adopt to cope with uncertainty. This research examines instead an experiential way of dealing with uncertainty and introduces a set of experimental methods showing how the experience of haptic softness can serve as a tool to deal with uncertainty.

实验研究研究如何人能不确定性通过软触觉感知应对

Human beings are constantly surrounded by uncertainty and change. The question arises how people cope with such uncertainty. To date, most research has ...

Examining Recall Memory in Infancy and Early Childhood Using the Elicited Imitation Paradigm

The ability to recall the past allows us to report on details of previous experiences, from the everyday to the significant. Because recall memory is commonly assessed using verbal report paradigms in adults, studying the development of this ability in preverbal infants and children proved challenging. Over the past 30 years, researchers have developed a non-verbal means of assessing recall memory known as the elicited or deferred imitation paradigm. In one variant of the procedure, participants are presented with novel three-dimensional stimuli for a brief baseline period before a researcher demonstrates a series of actions that culminate in an end- or goal-state. The participant is allowed to imitate the demonstrated actions immediately, after a delay, or both. Recall performance is then compared to baseline or to performance on novel control sequences presented at the same session; memory can be assessed for the individual target actions and the order in which they were completed. This procedure is an accepted analogue to the verbal report techniques used with adults, and it has served to establish a solid foundation of the nature of recall memory in infancy and early childhood. In addition, the elicited or deferred imitation procedure has been modified and adapted to answer questions relevant to other aspects of cognitive functioning. The broad utility and application of imitation paradigms is discussed, along with limitations of the approach and directions for future research.

The elicited imitation procedure was established to examine the development of recall memory in infancy and early childhood. This procedure has been widely used to establish a solid foundation of the nature of recall memory in infancy and early childhood.

使用仿引起范式检查召回内存在婴幼儿时期

The ability to recall the past allows us to report on details of previous experiences, from the everyday to the significant. Because recall memory is ...

Targeted Training of Ultrasonic Vocalizations in Aged and Parkinsonian Rats

Voice deficits are a common complication of both Parkinson disease (PD) and aging; they can significantly diminish quality of life by impacting communication abilities. 1, 2 Targeted training (speech/voice therapy) can improve specific voice deficits,3, 4 although the underlying mechanisms of behavioral interventions are not well understood. Systematic investigation of voice deficits and therapy should consider many factors that are difficult to control in humans, such as age, home environment, age post-onset of disease, severity of disease, and medications. The method presented here uses an animal model of vocalization that allows for systematic study of how underlying sensorimotor mechanisms change with targeted voice training. The ultrasonic recording and analysis procedures outlined in this protocol are applicable to any investigation of rodent ultrasonic vocalizations.
The ultrasonic vocalizations of rodents are emerging as a valuable model to investigate the neural substrates of behavior.5-8 Both rodent and human vocalizations carry semiotic value and are produced by modifying an egressive airflow with a laryngeal constriction.9, 10 Thus, rodent vocalizations may be a useful model to study voice deficits in a sensorimotor context. Further, rat models allow us to study the neurobiological underpinnings of recovery from deficits with targeted training.
To model PD we use Long-Evans rats (Charles River Laboratories International, Inc.) and induce parkinsonism by a unilateral infusion of 7 &mu;g of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle which causes moderate to severe degeneration of presynaptic striatal neurons (for details see Ciucci, 2010).11, 12 For our aging model we use the Fischer 344/Brown Norway F1 (National Institute on Aging).
Our primary method for eliciting vocalizations is to expose sexually-experienced male rats to sexually receptive female rats. When the male becomes interested in the female, the female is removed and the male continues to vocalize. By rewarding complex vocalizations with food or water, both the number of complex vocalizations and the rate of vocalizations can be increased (Figure 1).
An ultrasonic microphone mounted above the male's home cage records the vocalizations. Recording begins after the female rat is removed to isolate the male calls. Vocalizations can be viewed in real time for training or recorded and analyzed offline. By recording and acoustically analyzing vocalizations before and after vocal training, the effects of disease and restoration of normal function with training can be assessed. This model also allows us to relate the observed behavioral (vocal) improvements to changes in the brain and neuromuscular system.

Voice disorders are debilitating in aging and Parkinson disease. The ultrasonic vocalizations of rats, also affected by these conditions, can be used to study these voice disorders, their neural substrates, and the nature of functional recovery with behavioral intervention.

老年帕金森病大鼠的超声波发声的有针对性的培训

Voice deficits are a common complication of both Parkinson disease (PD) and aging; they can significantly diminish quality of life by impacting ...

Eliciting and Analyzing Male Mouse Ultrasonic Vocalization (USV) Songs

Mice produce ultrasonic vocalizations (USVs) in a variety of social contexts throughout development and adulthood. These USVs are used for mother-pup retrieval1, juvenile interactions2, opposite and same sex interactions3,4,5, and territorial interactions6. For decades, the USVs have been used by investigators as proxies to study neuropsychiatric and developmental or behavioral disorders7,8,9, and more recently to understand mechanisms and evolution of vocal communication among vertebrates10. Within the sexual interactions, adult male mice produce USV songs, which have some features similar to courtship songs of songbirds11. The use of such multisyllabic repertoires can increase potential flexibility and information they carry, as they can be varied in how elements are organized and recombined, namely syntax. In this protocol a reliable method to elicit USV songs from male mice in various social contexts, such as exposure to fresh female urine, anesthetized animals, and estrus females is described. This includes conditions to induce a large amount of syllables from the mice. We reduce recording of ambient noises with inexpensive sound chambers, and present a quantification method to automatically detect, classify and analyze the USVs. The latter includes evaluation of call-rate, vocal repertoire, acoustic parameters, and syntax. Various approaches and insight on using playbacks to study an animal&#39;s preference for specific song types are described. These methods were used to describe acoustic and syntax changes across different contexts in male mice, and song preferences in female mice.

Eliciting and Analyzing Male Mouse Ultrasonic Vocalization USV Songs

Mice produce a complex multisyllabic repertoire of ultrasonic vocalizations (USVs). These USVs are widely used as readouts for neuropsychiatric disorders. This protocol describes some of the practices we learned and developed to consistently induce, collect, and analyze the acoustic features and syntax of mouse songs.

引导和分析男性超声波声（USV）歌曲

Mice produce ultrasonic vocalizations (USVs) in a variety of social contexts throughout development and adulthood. These USVs are used for mother-pup ...