这项工作得到了资助的科学研究 (KAKENHI) 支持: 15H05724, 16H01491 和15H04289。

所有的实验都是按照国家遗传学研究所 (NIG) 的动物实验指南进行的, 所有的程序都是由 NIG 动物护理和使用委员会批准的 (批准 26-9)。
1. 动物
<ol><li>
选育
	<ol>
<li>通过交叉8个自交系 (MSM、HMI、BLG2、PGN2、KJR、冠心病、NJL 和 BFM/2), 创建野生衍生的异质股 (WHS)6只小鼠, 同时避免任何近亲繁殖 (兄弟姐妹或堂兄弟之间的交配)7。进行选择性育种, 从第三代开始 (G3), 通过选择显示更长的 "接触时间" (选定的组, 16 家庭交配对) 根据主动驯服测试 (见步骤 3.1)。</li>
		<li>通过从 G3 的随机交配产生非选定的控制组 (也16个交配对的家庭)。对于具有代表性的实验, 使用从选定的 (S1) 和非选定 (C1) 组 WHS 小鼠获得的小鼠, 以显示主动和被动驯服的模式使用三驯服测试 (见步骤 3.1-3.3)。对于实验, 每组使用 10 G15 小鼠 (S1 和 C1)。</li>
	</ol>
</li>
	<li>
代表主题动物
	<ol>
<li>在3周的时间里, 从父母那里戒掉所有的老鼠, 然后用窝的标准大小的塑料笼子, 把它们放在同性的群体里, 直到测试。维护所有动物在一个 NIG 动物设施在 12/12 h 光/黑暗的周期 (灯从06:00 到 18:00) 和恒定的温度 (23 @ 2 °c)。</li>
		<li>确保食物和水是可利用的广告随意。在光照期间对小鼠进行6周的试验, 分析它们在人类通常活跃的一天中与人类的相互作用。</li>
	</ol>
</li>
	<li>
处理协议
	<ol>
<li>使用用硅管覆盖的大镊子将老鼠转到新笼子里, 用尾巴轻轻地把它们捡起来。用镊子在断奶时将它们与父母分开, 并在实验过程中移动它们。在驯服测试之前, 所有的老鼠都不应该接受处理。</li>
		<li>在驯服测试之前把老鼠转移到实验室。</li>
		<li>在试验后保持雄性小鼠相互分离, 以避免雄性在 WHS 小鼠中的侵袭。</li>
	</ol>
</li>
</ol>2. 设备
<ol><li>使用开放领域设备 (400 x 400 x 400 毫米, 淡灰聚氯乙烯)。</li>
	<li>使用带有中性白色灯泡 (100 莱克丝) 的固定在打开字段上方450毫米的夹灯 (图 1) 来照亮打开的字段。</li>
	<li>使用三脚架 (图 1), 用商业上可用的紧凑型数码相机记录所有行为, 该摄像机固定在850毫米以上的开放场上。</li>
	<li>在无源驯服试验 (罕见反应) 的情况下, 在乳胶手套下戴上布手套, 防止手被咬伤。</li>
	<li>使用秒表测量测试周期。</li>
</ol>3. 测试
注: 前两项试验 (主动驯服和被动驯服) 分别进行了1分钟, 并进行了不到1分钟的手试试验。所有三项测试均在露天仪器中进行, 并使用数码相机进行记录。Iodophore 消毒程序应在所有 apparats 之前进行任何行为研究。
<ol><li>
程序,主动驯服测试
	<ol>
<li>打开数码相机并开始录制。</li>
		<li>用一双大镊子 (见步骤 1.3.1) 轻轻地用尾巴拿起鼠标, 把它从主笼转移到露天设备的中心。</li>
		<li>开始秒表。</li>
		<li>将左手放在器械的左下角。</li>
		<li>保持手掌朝上, 慢慢地和稳步地移动手指, 以确保鼠标把手和人连接起来。将所有四根手指从小指移动到食指的平均时间应约为0.5 秒, 连续重复运动周期。</li>
		<li>将手朝鼠标缓慢移动, 平均速度约为13厘米/秒, 从鼠标身体一侧的10厘米处停止。</li>
		<li>保持相同的距离, 如果鼠标试图离开。如果鼠标接近, 保持手在同一位置, 并继续移动手指轻微。如果鼠标试图爬上手臂, 用自由的手将它返回到现场。</li>
		<li>1分钟后完成试验, 取出手, 进行被动驯服试验。</li>
	</ol>
</li>
	<li>
程序,被动驯服测试
	<ol>
<li>按照与活动驯服测试相同的步骤, 但不要停止从鼠标的身体10厘米, 用指尖慢慢地触摸鼠标。</li>
		<li>把手指放在同一个位置, 联系鼠标, 直到鼠标移开。当鼠标试图打破接触, 跟随它, 以平均速度移动大约23厘米/秒, 并重新启动触摸。</li>
		<li>1分钟后完成测试, 立即重新启动秒表, 开始进行手动测试。</li>
	</ol>
</li>
	<li>
程序,手持式测试
	<ol>
<li>用一双大镊子抬起老鼠尾巴, 轻轻地放在左手掌上。把手紧贴在露天仪器的地板上。</li>
		<li>用左手拇指以0.5 秒/冲程的平均速度轻轻地描鼠标, 直到它离开手。</li>
		<li>重复步骤 3.3.1-3.3.2 3x。</li>
		<li>第三次试用后, 停止录像;所有驯服测试现已结束。</li>
	</ol>
</li>
	<li>
后实验程序
	<ol>
<li>对老鼠进行称量并计算出野外的粪便和尿量。</li>
		<li>通过耳拳分别标记鼠标。</li>
		<li>用0.2% 伏碘消毒液浸泡的纸巾擦拭仪器和实验手套。</li>
		<li>用干纸巾清洁器具。</li>
	</ol>
</li>
</ol>4. 行为分析
注: 视频录制使用 tanaMove 0.01 版 (Windows 7), 事件记录软件进行行为分析 (图 2)。该软件是免费提供 (补充材料 1)和我们的网站 (http://www.mgrl-lab.jp/MGRL_software.html)。一个对测试条件视而不见的人得分了所有的镜头。所有行为指数的持续时间以0.1 秒为增量。在评分过程中, 观察者在注意到给定的行为时按下了键盘上适当的字母 (参见步骤 4.1)。
<ol><li>
视频分析中按键的分配
	<ol>
<li>为每个行为索引分配键。对每个行为索引使用适当的键对于在步骤4.3 中自动汇总数据至关重要。 注意: 本研究中使用的作业如图 3所示。</li>
	</ol>
</li>
	<li>
视频分析
	<ol>
<li>分析每个视频记录3x 的不同行为指数 (图 3B)。自我梳理的行为没有得分。请确保计算机的内置扬声器已打开, 以便在视频中听到计时器声音。</li>
		<li>发射 tanaMove。</li>
		<li>将视频文件拖放到 tanaMove 窗口中进行分析。确认文件已加载, 并且数据文件 [. CSV]框中输出的 CSV 文件的路径和名称正确无误。</li>
		<li>单击 "就绪" 按钮。这将激活一个新窗口以输入指定的键。</li>
		<li>单击 "播放" 按钮启动视频文件。单击 "停止" 按钮暂停视频文件。</li>
		<li>当鼠标被观察到或接近手的时候按下键A , 包括站立嗅探。在活动驯服测试中, 当鼠标被观察到 "locomoting" 时, 按下键R , 而不向手方向移动。</li>
		<li>在六十年代听到计时器声音时, 按住键W至少0.3 秒 (在窗口中至少有三个间隔)。</li>
		<li>在左手进入视频后, 启动被动驯服测试的分析。当鼠标被观察到手和键R时, 按下键A , 当鼠标在被动驯服测试期间不向手方向移动时, 就会被观察到。</li>
		<li>在六十年代听到计时器声音时, 按住键W至少0.3 秒 (在窗口中至少有三个间隔)。</li>
		<li>当鼠标被观察到手上的 "停留" 时, 按下键T 。</li>
		<li>在手动测试之后, 单击 "停止" 按钮停止视频观察。</li>
		<li>单击 "后退" 按钮并键入 CSV 数据输出的新文件名。</li>
		<li>单击 "就绪" 按钮以激活新窗口以输入指定的键。</li>
		<li>单击 "播放" 按钮重新启动视频片段并启动对活动驯服测试的观察。</li>
		<li>按键T当鼠标被观察到 "接触" 人, 其中包括所有的自愿接触行为, 但不是偶然接触, 如在跳跃后降落在手掌, 在积极的驯服测试。</li>
		<li>在六十年代听到计时器声音时, 按住键W至少0.3 秒 (在窗口中至少有三个间隔)。</li>
		<li>当鼠标允许实验者在被动驯服测试时触摸它的身体时按键T 。</li>
		<li>在六十年代听到计时器声音时, 按住键W至少0.3 秒 (在窗口中至少有三个间隔)。</li>
		<li>单击 "停止" 按钮停止视频。</li>
		<li>单击 "后退" 按钮并键入 CSV 数据输出的新文件名。</li>
		<li>单击 "就绪" 按钮以激活新窗口以输入指定的键。</li>
		<li>单击 "播放" 按钮重新启动视频片段并启动对活动驯服测试的观察。</li>
		<li>当鼠标在活动驯服测试中被观察到 "跳跃" 时, 按下键J 。</li>
		<li>在六十年代听到计时器声音时, 按住键W至少0.3 秒 (在窗口中至少有三个间隔)。然后开始观察被动驯服试验。</li>
		<li>当鼠标在被动驯服测试中被观察到 "跳跃" 时, 按下键J 。</li>
		<li>在六十年代听到计时器声音时, 单击 "停止" 按钮停止视频观察。</li>
		<li>单击 "后退" 按钮并键入 CSV 数据输出的新文件名。</li>
	</ol>
</li>
	<li>
汇总数据
	<ol>
<li>将所有原始数据文件 (CSV) 保存在附带的2.xlsm文件 (补充材料 2) 的同一文件夹中。</li>
		<li>打开辅助 2.xlsm文件, 然后单击 "汇总" 按钮。</li>
		<li>将结果另存为单独的文件, 并使用它来进一步分析驯服行为。</li>
	</ol>
</li>
</ol>

<ol>
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作者没有什么可透露的。

代表的结果包括九行为指数从主动驯服, 被动驯服, 和停留在手的测试。对照组的分数比较显示驯服明显差异。结果表明, 三试验对小鼠驯服的测定是有效的。
动物的驯服水平很难从经验上量化。对鹿、小鼠和大鼠8、9、10驯服的评价方法进行了研究。然而, 这些方法并没有分别量化主动和被动驯服。在以前的研究中, 我们开发了三驯服测试, 以量化驯服在小鼠5。利用活性驯服试验的分数, 进行了选择性育种实验, 成功培育了6只活跃的小鼠。除了在主动驯服试验中更愿意接近和接触人类外, 在被动驯服试验中, 有选择地饲养的小鼠还可以耐受人的接触时间延长, 尽管该试验的结果未被用作选择的依据。这些结果表明, 这两项测试不是完全独立的, 并有一定的重叠。
为了准确测量小鼠对人类的先天驯服, 在驯服试验之前, 避免老鼠的处理是至关重要的。为了避免人类和老鼠之间的直接接触, 我们用大镊子盖上了硅管, 用尾巴把老鼠捡起来, 把它们转移到新的笼子5。硅管盖使转移较少痛苦对老鼠。
在每24小时的黑暗阶段, 我们没有进行驯服测试。然而, 测试光线和暗相是否影响驯服可能是很有趣的。由于开放领域的仪器被称为诱发小鼠的恐惧和压力反应, 由于这个原因经常被用来测量情感, 某些程度的驯服可能已被抑制在测试期间的恐惧/胁迫反应的小鼠。尽管我们能够在给定的测试条件下, 建立一些小鼠, 这些老鼠在实验中表现出高水平的活性驯服, 但未来的研究应该设法减少老鼠在测试过程中的恐惧和压力, 例如通过进行测试在黑暗阶段或通过 habituating 老鼠到仪器在测试之前。
此外, 对试验动物 (选择的或非选定的) 的状态进行驯服测试, 可以使测试更可靠8。
值得注意的是, 三的测试可能适用于实验室和野鼠菌株5。由于这些试验可以应用于野生菌株, 我们能够建立一个小鼠种群展示高活性驯服使用一个基因异构的股票下降八野生老鼠菌株6。
驯服测试结果通过视频评分进行量化。为了确保一致性, 一个人分析了目前研究中的选择性育种实验的所有视频数据。然而, 这种方法要求观察员是一个专家谁可以产生可靠和可重复的结果, 所以应注意正确训练视频记分员。驯服在小鼠身上的未来工作可能会利用自动分析软件取代人的观察者, 这种方法在以前的社会行为研究中使用过11。

在驯化过程中选择育种改变了驯养动物的特性, 使其更好地适应人类生活1,2,3。"驯服" 是家养动物观察到的一种常见的行为特征。动物驯服的选择性育种研究揭示了它们在驯化过程中的表型变化。在一项著名的研究中, 狐狸有选择地繁殖为驯服和一组狐狸, 展示了高水平的驯服建立了2。狐狸还显示了其他表型变化, 包括形态学, 繁殖, 大衣颜色和生理特性。驯服的选择性育种是一种流行的实验动物, 可以帮助揭示在驯化过程中各种表型变化的机制。
驯服有二个行为组分: 活跃方法对人 (活跃驯服) 和减少在人的退避 (被动驯服)4。在以前的一项研究中, 我们建立了三项行为测试来表征小鼠的驯服水平: 主动驯服, 被动驯服, 以及手头的测试5。第一个测试的目的是通过测量其自发方法对人类的潜伏期和与人类手接触的时间, 来量化老鼠的活动驯服。第二个测试是为了量化被动驯服通过测量老鼠容忍被人的手接触的时间。该试验旨在量化行为避免被人的手轻轻抚摸。这三项评估只需要一个露天设备、一台数码相机和一双大镊子。它们可以按顺序在3分钟内完成, 以便每天收集大量数据。
在前一项研究中, 我们用一系列的近交系小鼠菌株对驯服行为进行了表征和比较:10 株野生菌株, 一只日本奇特的老鼠源菌株, 六株实验室菌株5。大多数驯化菌株的被动驯服明显高于野生菌株, 但活性驯服在两组间没有差异。从这些结果, 我们得出结论, 驯化的老鼠是实验室菌株的创始人主要选择他们的被动驯服, 但不为活跃的驯服, 在他们的繁殖历史。
我们接下来进行选择性育种与八野生小鼠菌株, 以产生一个基因异构人口6。我们选择了在主动驯服试验中得分高的动物, 并成功地增加了他们的活跃驯服在连续世代, 与对照组相比。这一结果表明, 试验可靠地测量小鼠驯服。
在这里, 我们描述了使用三行为测试测量行为指数的综合方法。首先, 我们解释了设置和必要的设备。接下来, 我们详细介绍了运行实验的分步过程, 通过视频收集数据, 并使用我们开发的免费软件分析所说的数据。最后, 给出了对照组选择性饲养小鼠池的典型结果。

<table border="0" cellpadding="0" cellspacing="0" width="818">
	<tbody>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">open-field apparatus</td>
			<td style="width:169px;">O&#8217;Hara &#38; Co. Ltd.</td>
			<td style="width:151px;">OF-3102</td>
			<td style="width:283px;">400&#215;400&#215;400 mm, grayish polyvinylchloride</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">compact digital camera</td>
			<td>Ricoh Company, Ltd.</td>
			<td>CX-5</td>
			<td>production in the company is over</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">latex pure gloves</td>
			<td>AS ONE</td>
			<td>BioLab, IKG-8002</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">stopwatch</td>
			<td>SEIKO</td>
			<td>SSBJ018</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Iodophor sanitizer</td>
			<td>Ecolab G.K. (CLEA Japan)</td>
			<td>CL-4123</td>
			<td>Mikro Klene</td>
		</tr>
	</tbody>
</table>

measuring active and passive tameness separately in mice

家养的动物, 如狗和实验鼠, 显示出高水平的驯服, 这对于人类很容易处理它们很重要。驯服有两个行为成分: 不情愿避免人类 (被动驯服) 和一种接近人类的动机 (主动驯服)。为了量化这些组分在小鼠, 我们以前开发了行为测试为活跃驯服, 被动驯服和愿意停留在人的手, 每设计在3分钟之内完成。所获得的数据用于选择性育种, 每代分析大量小鼠。活动驯服测试测量鼠标的移动对人的手和它参与的接触。被动驯服测试测量老鼠容忍人类接触的时间长短。在手持测试中, 鼠标放在人的手上, 用手的拇指慢慢地接触;动物保持在手上的时间长短被测量。在这里, 我们描述的测试设置和仪器, 解释程序, 并讨论适当的数据分析。最后, 我们解释了如何解释结果。

我们以前进行了选择性育种的活性驯服在 miceandsuccessfully 生产的小鼠, 显示高水平的积极驯服6。图 4显示了15代 (G15) 中选定和非选定的小鼠组的驯服测试的代表性结果。所选组的小鼠在主动驯服试验中得分较高, 与对照鼠相比, 与人手接触的时间明显延长。这种接触持续时间是一种用于选择性育种的指标。两组在航向、运动或跳跃持续时间上没有显著差异 (图 4A,表 1), 但选定的小鼠在被动驯服测试中得分较高, 使人类接触时间大大延长。比控制鼠做的。此外, 选定的老鼠的运动和跳跃持续时间明显缩短 (图 4B,表 1)。最后, 选定的老鼠在人类手上的停留时间明显长于在手试验中的控制鼠 (图 4C,表 1)。这些结果表明, 选择性饲养的小鼠比控制小鼠更有教养。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/58048/58048fig1.jpg"> 图 1: 驯服测试仪.这些面板显示的仪器从不同的角度 (左和右)。该设置包括一个开放的领域, 三脚架, 剪辑灯, 数码相机, 秒表, 耳冲床, 大镊子, 和一个塑料烧杯含有0.2% 碘伏消毒剂。在测试期间, 剪辑指示灯已打开。三脚架高度调整, 使数码相机可以捕捉整个空间内的仪器。测试后立即进行耳部冲孔和体重测量。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58048/58048fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/58048/58048fig2.jpg"> 图 2: 通过视频评分测量行为指数.使用软件 tanaMove 对驯服测试中的行为指标进行了评分。此图显示了一个在活动驯服测试开始后的17.7 秒的视频录制中的静止镜头。鼠标与实验者接触, 所以观察者记录了键T, 如 tanaMove 的编辑窗口所示。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58048/58048fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/58048/58048fig3.jpg"> 图 3: 使用 tanaMove 对视频数据进行分析.(A) 每个行为索引都分配给键盘上的一个键。(B) 进行主动和被动的驯服试验, 每1分钟进行一次, 然后进行持续试验。观察指标按时间顺序测定, 1 至3。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58048/58048fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/58048/58048fig4.jpg"> 图 4: 驯服测试的代表性结果.对于代表性实验, 我们使用了从选定的 (S1) 和非选择 (C1) 组 WHS 的小鼠, 以显示主动和被动驯服使用三驯服测试的模式。实验中, 使用了每组 10 G15 小鼠 (S1 和 C1)。(A) 这些面板显示了活动驯服测试的结果。四行为指数被比较的选择和对照组: 标题, 运动, 接触和跳跃。(B) 这些小组显示被动驯服测试的结果。四行为指数被比较的选择和对照组: 标题, 运动, 接受和跳跃。(C) 本小组展示了手头测试的结果。在选择和控制组之间比较了一个索引: 停留在手上。所有值均为三项试验的平均值。误差线表示 SEM.<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58048/58048fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td>驯服测试</td>
			<td>行为指数</td>
			<td>组</td>
			<td>夏皮罗-Wilk 测试</td>
			<td>曼-惠特尼 U 测试</td>
		</tr>
<tr>
<td rowspan="8">主动驯服试验</td>
			<td rowspan="2">标题</td>
			<td>控制</td>
			<td>0.3991</td>
			<td rowspan="2">0.5922</td>
		</tr>
<tr>
<td>选择</td>
			<td>0.4907</td>
		</tr>
<tr>
<td rowspan="2">运动</td>
			<td>控制</td>
			<td>0.3832</td>
			<td rowspan="2">0.0853</td>
		</tr>
<tr>
<td>选择</td>
			<td>0.3599</td>
		</tr>
<tr>
<td rowspan="2">接触</td>
			<td>控制</td>
			<td>< 0.0001</td>
			<td rowspan="2">0.0049</td>
		</tr>
<tr>
<td>选择</td>
			<td>0.1336</td>
		</tr>
<tr>
<td rowspan="2">跳</td>
			<td>控制</td>
			<td>< 0.0001</td>
			<td rowspan="2">0.2105</td>
		</tr>
<tr>
<td>选择</td>
			<td>―</td>
		</tr>
<tr>
<td rowspan="8">被动驯服试验</td>
			<td rowspan="2">标题</td>
			<td>控制</td>
			<td>0.0408</td>
			<td rowspan="2">0.0600</td>
		</tr>
<tr>
<td>选择</td>
			<td>0.7725</td>
		</tr>
<tr>
<td rowspan="2">运动</td>
			<td>控制</td>
			<td>0.1470</td>
			<td rowspan="2">0.0373</td>
		</tr>
<tr>
<td>选择</td>
			<td>0.4356</td>
		</tr>
<tr>
<td rowspan="2">接受</td>
			<td>控制</td>
			<td>0.5834</td>
			<td rowspan="2">0.0023</td>
		</tr>
<tr>
<td>选择</td>
			<td>0.7742</td>
		</tr>
<tr>
<td rowspan="2">跳</td>
			<td>控制</td>
			<td>0.0048</td>
			<td rowspan="2">0.0108</td>
		</tr>
<tr>
<td>选择</td>
			<td>―</td>
		</tr>
<tr>
<td rowspan="2">停留在手测试</td>
			<td rowspan="2">住</td>
			<td>控制</td>
			<td>0.6683</td>
			<td rowspan="2">0.0003</td>
		</tr>
<tr>
<td>选择</td>
			<td>0.1897</td>
		</tr>
</tbody></table>表 1: 比较所选和对照组行为指数的统计分析结果.Wilk 和曼-惠特尼 U 型测试的p值被显示出来。鉴于某些行为指数具有非正态分布, 我们使用曼-惠特尼 U 型测试来检验所选和对照组之间的差异。破折号 (-) 意味着不能计算统计, 因为所有的人都表现出零分。

Watch this Scientific Journal Video about Measuring Active and Passive Tameness Separately in Mice at JoVE.com

Measuring Active and Passive Tameness Separately in Mice

驯服在动物包括减少退避反应对人 (被动驯服) 和增加活跃方法对人 (活跃驯服)。在这里, 我们描述了三行为测试 (主动驯服, 被动驯服和保持手测试) 的详细协议, 以单独测量小鼠的主动和被动驯服。

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 ...

measuring-active-and-passive-tameness-separately-in-mice

Nanyang Technological University

Research

JoVE Journal

Behavior

7.0K 次观看.  National Institute of Genetics. 驯服在动物包括减少退避反应对人 (被动驯服) 和增加活跃方法对人 (活跃驯服)。在这里, 我们描述了三行为测试 (主动驯服, 被动驯服和保持手测试) 的详细协议, 以单独测量小鼠的主动和被动驯服。

视频: Measuring Active and Passive Tameness Separately in Mice

The Successive Alleys Test of Anxiety in Mice and Rats

The plus-maze was derived from the early work of Montgomery. He observed that rats tended to avoid the open arms of a maze, preferring the enclosed ones. Handley, Mithani and File et al. performed the first studies on the plus-maze design we use today, and in 1987 Lister published a design for use with mice. 
Time spent on, and entries into, the open arms are an index of anxiety; the lower these indices, the more anxious the mouse is. Alternatively, a mouse that spends most of its time in the closed arms is classed as anxious. 
One of the problems of the plus-maze is that, while time spent on, and entries into, the open arms is a fairly unambiguous measure of anxiety, time in the central area is more difficult to interpret, although time spent here has been classified as &#x201C;decision making&#x201D;. In many tests central area time is a considerable part of the total test time.
Shepherd et al. produced an ingenious design to eliminate the central area, which they called the &#x201C;zero maze&#x201D;. However, although used by several groups, it has never been as widely adopted as the plus-maze. 
In the present article I describe a modification of the plus-maze design that not only eliminates the central area but also incorporates elements from other anxiety tests, such as the light-dark box and emergence tests. It is a linear series of four alleys, each having increasing anxiogenic properties. It has given similar results to the plus-maze in general. Although it may not be more sensitive than the plus-maze (more data is needed before a firm conclusion can be reached on this point), it provides a useful confirmation of plus-maze results which would be useful when, for example, only a single example of a mutant mouse was available, as, for example, in ENU-based mutagenesis programs.

The plus-maze measures anxiety-like behaviour in rodents. There are two opposite closed and two opposite open arms; anxious rodents avoid the open arms. The central area is neither completely open nor closed, so time spent here is ambiguous and difficult to interpret. Here a modification of the plus-maze protocol eliminating this area is described.

在小鼠和大鼠连续的小巷测试焦虑

The plus-maze was derived from the early work of Montgomery. He observed that rats tended to avoid the open arms of a maze, preferring the enclosed ones.  ...

科研

行为学

Trace Fear Conditioning in Mice

In this experiment we present a technique to measure learning and memory. In the trace fear conditioning protocol presented here there are five pairings between a neutral stimulus and an unconditioned stimulus. There is a 20 sec trace period that separates each conditioning trial. On the following day freezing is measured during presentation of the conditioned stimulus (CS) and trace period. On the third day there is an 8 min test to measure contextual memory. The representative results are from mice that were presented with the aversive unconditioned stimulus (shock) compared to mice that received the tone presentations without the unconditioned stimulus. Trace fear conditioning has been successfully used to detect subtle learning and memory deficits and enhancements in mice that are not found with other fear conditioning methods. This type of fear conditioning is believed to be dependent upon connections between the medial prefrontal cortex and the hippocampus. One current controversy is whether this method is believed to be amygdala-independent. Therefore, other fear conditioning testing is needed to examine amygdala-dependent learning and memory effects, such as through the delay fear conditioning.

In the following experiment we describe a protocol for trace fear conditioning in mice. This type of associative memory includes a trace period that separates the neutral stimulus and the unconditioned stimulus.

跟踪恐惧条件小鼠

In this experiment we present a technique to measure learning and memory. In the trace fear conditioning protocol presented here there are five pairings ...

Measuring Attentional Biases for Threat in Children and Adults

Investigators have long been interested in the human propensity for the rapid detection of threatening stimuli. However, until recently, research in this domain has focused almost exclusively on adult participants, completely ignoring the topic of threat detection over the course of development. One of the biggest reasons for the lack of developmental work in this area is likely the absence of a reliable paradigm that can measure perceptual biases for threat in children. To address this issue, we recently designed a modified visual search paradigm similar to the standard adult paradigm that is appropriate for studying threat detection in preschool-aged participants. Here we describe this new procedure. In the general paradigm, we present participants with matrices of color photographs, and ask them to find and touch a target on the screen. Latency to touch the target is recorded. Using a touch-screen monitor makes the procedure simple and easy, allowing us to collect data in participants ranging from 3 years of age to adults. Thus far, the paradigm has consistently shown that both adults and children detect threatening stimuli (e.g., snakes, spiders, angry/fearful faces) more quickly than neutral stimuli (e.g., flowers, mushrooms, happy/neutral faces). Altogether, this procedure provides an important new tool for researchers interested in studying the development of attentional biases for threat.

Here we describe a touch-screen visual search paradigm that can be used to study threat detection across the lifespan. The paradigm has already been used in various studies demonstrating that both children and adults detect threatening stimuli like snakes, spiders, and angry faces faster than non-threatening stimuli.

测量注意偏向对威胁儿童和成人

Investigators have long been interested in the human propensity for the rapid detection of threatening stimuli. However, until recently, research in this ...

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal&#8217;s behavioral profile can be investigated in one single test.
The apparatus consists of a box, representing the &#8216;protected&#8217; area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the &#8216;unprotected&#8217; area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related- and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals&#8217; adaptive capacities.
Due to testing order effects in a behavioral test battery, na&#239;ve animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress.

This protocol describes the modified hole board, which is a behavioral test set-up that comprises the characteristics of an open field and a traditional hole board. This set-up enables the differential analysis of unconditioned behavior of small laboratory mammals as well as the analysis of cognitive abilities.

修改后的孔板 - 测量行为，认知和社会互动小鼠和大鼠

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

Testing for Odor Discrimination and Habituation in Mice

This video demonstrates a technique to establish the presence of a normally functioning olfactory system in a mouse. The test helps determine whether the mouse can discriminate between non-social odors and social odors, whether the mouse habituates to a repeatedly presented odor, and whether the mouse demonstrates dishabituation when presented with a novel odor. Since many social behavior tests measure the experimental animal&#8217;s response to a familiar or novel mouse, false positives can be avoided by establishing that the animals can detect and discriminate between social odors. There are similar considerations in learning tests such as fear conditioning that use odor to create a novel environment or olfactory cues as an associative stimulus. Deficits in the olfactory system would impair the ability to distinguish between contexts and to form an association with an olfactory cue during fear conditioning. 
In the odor habitation/dishabituation test, the mouse is repeatedly presented with several odors. Each odor is presented three times for two minutes. The investigator records the sniffing time directed towards the odor as the measurement of olfactory responsiveness. A typical mouse shows a decrease in response to the odor over repeated presentations (habituation). The experimenter then presents a novel odor that elicits increased sniffing towards the new odor (dishabituation). After repeated presentation of the novel odor the animal again shows habituation. This protocol involves the presentation of water, two or more non-social odors, and two social odors. In addition to reducing experimental confounds, this test can provide information on the function of the olfactory systems of new knockout, knock-in, and conditional knockout mouse lines.

This manuscript describes a protocol to examine the olfactory system of rodents. The olfactory habituation/dishabituation test will allow investigators to determine whether a mouse habituates to a repeatedly presented odor and whether the mouse demonstrates dishabituation when presented a novel odor.

在小鼠试验的气味辨别和习惯化

This video demonstrates a technique to establish the presence of a normally functioning olfactory system in a mouse. The test helps determine whether the ...

Continuous Noninvasive Measuring of Crayfish Cardiac and Behavioral Activities

A crayfish is a pivotal aquatic organism that serves both as a practical biological model for behavioral and physiological studies of invertebrates and as a useful biological indicator of water quality. Even though crayfish cannot directly specify the substances that cause water quality deterioration, they can immediately (within a few seconds) warn humans of water quality deterioration via acute changes in their cardiac and behavioral activities.
In this study, we present a noninvasive method that is simple enough to be implemented under various conditions due to a combination of simplicity and reliability in one model.
This approach, in which the biological organisms are implemented into environmental evaluation processes, provides a reliable and timely alarm for warning of and preventing acute water deterioration in an ambient environment. Therefore, this noninvasive system based on crayfish physiological and ethological parameter recordings was investigated for the detection of changes in an aquatic environment. This system is now applied at a local brewery for controlling&#160;quality of the water used for beverage production, but it can be used at any water treatment and supply facility for continuous, real-time water quality evaluation and for regular laboratory investigations of crayfish cardiac physiology and behavior.

This article presents a noninvasive biomonitoring system for the continuous recording and analyses of crayfish cardiac and locomotor activities. This system consists of a near-infrared optical sensor, a video-tracking module, and software for evaluating crayfish heartbeats that reflects its physiological condition and characterizes crayfish behavior during heartbeat fluctuations.

小龙虾心脏的连续无创测量与行为活动

A crayfish is a pivotal aquatic organism that serves both as a practical biological model for behavioral and physiological studies of invertebrates and as ...

Assessing the Autonomic and Behavioral Effects of Passive Motion in Rats using Elevator Vertical Motion and Ferris-Wheel Rotation

The overall goal of this study is to assess the autonomic and behavioral effects of passive motion in rodents using the elevator vertical motion and Ferris-wheel rotation devices. These assays can help confirm the integrity and normal functioning of the autonomic nervous system. They are coupled to quantitative measures based on defecation counting, open-field examination, and balance beam crossing. The advantages of these assays are their simplicity, reproducibility, and quantitative behavioral measures. The limitations of these assays are that the autonomic reactions could be epiphenomena of non-vestibular disorders and that a functioning vestibular system is required. Examination of disorders such as motion sickness will be greatly aided by the detailed procedures of these assays.

Protocols are presented to assess the autonomic and behavioral effects of passive motion in rodents using elevator vertical motion and Ferris-wheel rotation.

使用电梯垂直运动和摩天轮旋转评估大鼠被动运动的自主性和行为效应

The overall goal of this study is to assess the autonomic and behavioral effects of passive motion in rodents using the elevator vertical motion and ...

Assessment of Memory Function in Pilocarpine-induced Epileptic Mice

Cognitive impairment is one of the most common comorbidities in temporal lobe epilepsy. To recapitulate epilepsy-associated cognitive decline in an animal model of epilepsy, we generated pilocarpine-treated chronic epileptic mice. We present a protocol for three different behavioral tests using these epileptic mice: novel object location (NL), novel object recognition (NO), and pattern separation (PS) tests to evaluate learning and memory for places, objects, and contexts, respectively. We explain how to set the behavioral apparatus and provide experimental procedures for the NL, NO, and PS tests following an open field test that measures the animals&#8217; basal locomotor activities. We also describe the technical advantages of the NL, NO, and PS tests with respect to other behavioral tests for assessing memory function in epileptic mice. Finally, we discuss possible causes and solutions for epileptic mice failing to make 30 s of good contact with the objects during the familiarization sessions, which is a critical step for successful memory tests. Thus, this protocol provides detailed information about how to assess epilepsy-associated memory impairments using mice. The NL, NO, and PS tests are simple, efficient assays that are appropriate for the evaluation of different kinds of memory in epileptic mice.

This article presents experimental procedures for assessing memory impairments in pilocarpine-induced epileptic mice. This protocol can be used to study the pathophysiologic mechanisms of epilepsy-associated cognitive decline, which is one of the most common comorbidities in epilepsy.

皮洛卡平诱导癫痫小鼠记忆功能评估

Cognitive impairment is one of the most common comorbidities in temporal lobe epilepsy. To recapitulate epilepsy-associated cognitive decline in an animal ...

Tactile Semiautomatic Passive-Finger Angle Stimulator (TSPAS)

Passive tactile perception is the ability to passively and statically perceive stimulus information coming from the skin; for example, the ability to sense spatial information is the strongest in the skin on the hands. This ability is termed tactile spatial acuity, and is measured by the tactile threshold or discrimination threshold. At present, the two-point threshold is extensively used as a measure of tactile spatial acuity, although many studies have indicated that critical deficits exist in two-point discrimination. Therefore, a computer-controlled tactile stimulus system was developed, the tactile semiautomated passive-finger angle stimulator (TSPAS), using the tactile angle discrimination threshold as a new measure for tactile spatial acuity. The TSPAS is a simple, easily operated system that applies raised angle stimuli to a subject&#39;s passive fingerpad, while controlling movement speed, distance, and contact duration. The components of the TSPAS are described in detail as well as the procedure to calculate the tactile angle discrimination threshold.

Tactile Semiautomatic Passive-Finger Angle Stimulator TSPAS

Presented is the tactile semiautomated passive-finger angle stimulator TSPAS, a new way to assess tactile spatial acuity and tactile angle discrimination using a computer-controlled tactile stimulus system that applies raised angle stimuli to a subject&#39;s passive fingerpad, while controlling for movement speed, distance, and contact duration.

触觉半自动被动手指角刺激器 （TSPAS）

Passive tactile perception is the ability to passively and statically perceive stimulus information coming from the skin; for example, the ability to ...

Chronic Stress Shifts Effort-Related Choice Behavior in a Y-Maze Barrier Task in Mice

Mood disorders, including major depressive disorder, can be precipitated by chronic stress. The Y-maze barrier task is an effort-related choice test that measures motivation to expend effort and obtain reward. In mice, chronic stress exposure significantly impacts motivation to work for a higher value reward when a lesser value reward is freely available compared to unstressed mice. Here we describe the chronic corticosterone administration paradigm, which produces a shift in effortful responding in the Y-maze barrier task. In the Y-maze task, one arm contains 4 food pellets, while the other arm contains only 2 pellets. After mice learn to select the high reward arm, barriers with progressively increasing height are then introduced into the high reward arm over multiple test sessions. Unfortunately, most chronic stress paradigms (including corticosterone and social defeat) were developed in male mice and are less effective in female mice. Therefore, we also discuss chronic non-discriminatory social defeat stress (CNSDS), a stress paradigm we developed that is effective in both male and female mice. Repeating results with multiple distinct chronic stressors in male and female mice combined with increased usage of translationally relevant behavior tasks will help to advance the understanding of how chronic stress can precipitate mood disorders.

The Y-maze barrier task is a behavior test that examines motivation to expend effort for reward. Here, we discuss testing multiple well-validated chronic stressors including chronic corticosterone and social defeat stress with this behavior, as well as the novel chronic non-discriminatory social defeat stress (CNSDS), which is effective in females.

在小鼠的 Y-Maze 障碍任务中，慢性应力转移与工作量相关的选择行为

Mood disorders, including major depressive disorder, can be precipitated by chronic stress. The Y-maze barrier task is an effort-related choice test that ...