这项工作得到了中国国家自然科学基金委员会（61327902-6对W.王和81801318对丁F.）的支持。我们感谢西格丽德·维塞博士建立了SF实验系统，并亲切地提供了技术细节。我们感谢迈肯·内德加德博士对相关实验的启发性评论。

该协议经同济医院、同济医学院、华中科技大学动物护理与使用委员会批准。
1. 老鼠筛选和实验准备
<ol><li>在整个实验中，选择体重为20-28克的野生型成年小鼠（8-10周大）雄性小鼠。 注：野生型C57BL/6小鼠来自中国湖北实验室动物研究中心。</li>
	<li>随机将所有小鼠分配给CSF和对照组。每个笼子里住3-5只老鼠，以避免社会隔离压力。控制笼中容纳的老鼠数量与配对CSF笼中的老鼠数量相匹配。 注：同一组笼子中的老鼠被集中起来进行后续行为实验。</li>
	<li>将控制笼与CSF笼子放在同一个房间内，以保持周围环境和劳动效果的一致。</li>
	<li>使用耳标来标记和标记每组耳朵上的鼠标，用于监测目的。</li>
	<li>保持21-23°C至35%-60%之间的环境温度和湿度。</li>
	<li>保持环境环境在12小时的光-暗周期（上午8：00-晚上8：00点开灯，晚上8：00-上午8：00光关），以避免对小鼠正常睡眠节奏的偏差影响。</li>
	<li>当研究人员在建模室时，尽量减少噪音和干扰。</li>
	<li>为老鼠提供充足的食物和水。在水瓶上使用带球阀尖的长喷嘴，防止平台运动时漏水。用弹簧将水瓶固定在笼子顶部，以避免在转子运行过程中瓶子脱位。</li>
</ol>2. 轨道转子的准备和设置
<ol><li>准备一个电动控制的轨道转子与扩大的平台（67厘米x110厘米），其中最多可以放置10个笼子。</li>
	<li>在由程序计时器控制的光上阶段（上午 8：00-8：00）期间设置轨道转子，此时小鼠将展示其大部分日常睡眠。</li>
	<li>设置轨道转子，速度为 110 rpm，重复周期为 10 s-on，110 s-off，由固态定时器控制。 注：平台的负载容量为 50 kg。转子地平线振动的固定振幅为2.5厘米。</li>
	<li>用厚厚的弹簧将CSF笼子固定在转子平台顶部，以防止在平台旋转时笼子脱位。</li>
</ol>3. 慢性睡眠碎片建模和监测
<ol><li>在实验前将CSF和对照小鼠的笼子放入建模室一周，让小鼠适应环境环境。</li>
	<li>在建模开始时，确保所有小鼠在轨道旋转期间都能自由获得食物和水。</li>
	<li>在建模开始时，至少观察1小时，以确保轨道转子在齿轮上运行。</li>
	<li>在建模期间，检查轨道转子是否正常运行，小鼠每2天进行一次条件检查，以确保小鼠有足够的食物和水。每周更换笼子的床上用品。</li>
	<li>在建模期间，更换床上用品时，每周上午 8：00 给小鼠称重。将体重显著减轻的小鼠从建模中取出，也从实验组中取出。 注：显著减肥的定义是体重低于20克持续2周。</li>
	<li>在整个建模过程中，将侵略者（如果有的话）从笼子里移走，也从实验组中取出。</li>
	<li>建模终止后，继续维持和喂养原房间内的小鼠。</li>
</ol>4. 莫里斯水迷宫（MWM）测试
<ol><li>测试准备<ol>
<li>准备装满温水（20-23°C）的圆形水箱的装置。</li>
		<li>将四个形状和颜色不同的标志悬挂在坦克周围的窗帘上，以四个象限方向作为远视参考。添加奶粉使水显得不透明。</li>
		<li>在西南象限的中间定位一个平台。</li>
	</ol>
</li>
	<li>培训测试<ol>
<li>在为期5天的训练期间，每天上午8：00至12：00对小鼠进行连续四次试验。</li>
		<li>在四个试验中，将每只鼠标释放到面向侧壁的水中，这是四个象限之一。在每一次试验中，让老鼠游60年代才能找到平台。如果鼠标在 60 年代内无法到达平台，则引导它到平台，并在那里停留 15 s。</li>
		<li>使用视频跟踪系统自动记录小鼠的逃生延迟，以查找隐藏的平台。</li>
	</ol>
</li>
	<li>探头测试<ol>
<li>经过5天的训练后，于第六天进行探针测试。</li>
		<li>删除平台。从东北象限释放每只鼠标，并允许它游泳60s</li>
		<li>使用视频跟踪系统自动记录小鼠的跟踪数据。</li>
	</ol>
</li>
</ol>5. 新颖对象识别 （NOR） 测试
<ol><li>熟悉的阶段<ol>
<li>将小鼠按顺序放置在一个水箱中（长30厘米，宽28厘米，高35厘米），其中包含两个副本的物体（A1和A2）。让小鼠自由探索（每次试验10分钟）。</li>
		<li>使用视频跟踪系统自动记录小鼠的跟踪数据。</li>
	</ol>
</li>
	<li>测试阶段<ol>
<li>在熟悉阶段延迟 1 小时后进行测试试验。将其中一个原始对象替换为油箱中的新对象（"小说"），使另一个对象保持不变。将小鼠送回水箱，每次试用5分钟。</li>
		<li>使用视频跟踪系统自动记录每个鼠标在探索每个对象时花费的时间。 注：物体的探索是由舔、嗅、咀嚼或移动振动决定的，同时将鼻子朝向物体，距离物体不到1厘米。歧视索引 （DI） 是用方程 （TN + TF） / （TN + TF） 计算的，其中 TN = 探索"新颖"对象的时间和 TF = 探索"熟悉"对象所花费的时间。</li>
	</ol>
</li>
</ol>6. 开放式现场测试（OFT）
<ol><li>准备坦克的装置（30厘米×28厘米×35厘米）。</li>
	<li>在测试期间，将每个鼠标放入油箱中心，并允许它自由探索 5 分钟。每次试验后用 75% 的乙醇清洁油箱，以避免前一个鼠标的剩余效果。</li>
	<li>使用视频跟踪系统自动记录小鼠的跟踪数据。</li>
</ol>7. 强迫游泳测试（FST）
<ol><li>准备一个开放的圆柱形容器的仪器，其中包含水（20-23°C）是15厘米深。</li>
	<li>在测试期间，将每个鼠标放入气缸中，并允许其在那里停留 6 分钟。</li>
	<li>使用视频跟踪系统自动记录每个鼠标测试的最后 4 分钟的不动时间。 注：当老鼠停止挣扎和漂浮在水中时，它决心不动，只做必要的运动，以保持它的头在水面上。</li>
</ol>8. 数据分析
<ol></ol>

作者宣称他们没有相互竞争的经济利益。

当前协议中的关键步骤包括根据研究目的设置具有优化参数的睡眠碎裂机，并在整个建模过程中保持小鼠在舒适和安静的生活环境中。决定适当的时间来中断或停止睡眠分裂，并为这些小鼠安排行为测试也是至关重要的。与其他睡眠操作模型一样，在具有受控光周期且没有所有可能的不必要干扰的专用房间执行协议非常重要。应努力避免引起噪音，尽量减少研究人员检查、补充食物和供水、更换?...

睡眠障碍在睡眠干扰状况患者和有睡眠干扰事件的健康人中越来越常见。据观察，患有神经退行性疾病、慢性疼痛、情绪压力、呼吸系统疾病、泌尿系统疾病等的患者，通常抱怨睡眠不愉快的经历1、2、3、4、5。阻塞性睡眠呼吸暂停（OSA）、睡眠中的周期性肢体运动（PLMS）、睡眠维持失眠等睡眠障碍是最常见的原因，诱发睡眠分裂6、7。在发达国家，OSA在成人人口中的患病率超过5%至9%，在8、9、10岁儿童中患病率超过2%。同时，由于智能手机的过度使用、不规则的睡眠习惯、恼人的噪音和工作职责（如照顾者的夜班）而遭受睡眠障碍的健康人群比例越来越大。睡眠被认为是重要的脑废物清除11，12，记忆巩固13，14，代谢平衡15，16，许多其他生理过程。然而，长期睡眠障碍是否会导致健康人不可逆转的发病机制变化，以及它是未来几年内发展中枢神经系统疾病（如神经退行性疾病）的病因还是促成因素，仍在很大程度上还不得而知。我们的目标是报告一个实验模型，在经过2个月的睡眠分裂治疗后，在年轻的野生小鼠中产生稳定和明显的认知缺陷和焦虑样的行为。此模型将用于回答上述科学问题。
睡眠障碍被列为发展阿尔茨海默氏症（AD）或痴呆症的潜在危险因素。康等人首次发现并描述了AD病理学恶化6小时急性睡眠剥夺17。此后，许多其他研究报告说，睡眠不足或分裂可能加剧转基因AD小鼠模型18，19，20的发病机理。然而，很少有研究人员研究过幼小野鼠睡眠障碍的后果：即睡眠障碍是否会引起年轻野性小鼠的AD样行为或病理变化。在我们最近的出版物中，我们报告说，2个月的睡眠分裂导致明显的空间记忆缺陷和焦虑样的行为，以及增加细胞内淀粉样β（A+）积累在皮层和海马在2-3个月大的野生型小鼠21。我们还观察到内体-自噬体溶酶体通路标记和微胶质活化的表达水平改变，这与APP/PS1小鼠21、22中报告的病理变化相似。
这份提交的睡眠碎片（SF）协议经辛顿等人23日验证，并经李等人24日修改。简言之，在110 rpm的轨道转子振动，在光打开阶段（上午8：00-晚上8：00）每2分钟中断睡眠10秒。该模型中的睡眠结构变化以前以电生理睡眠记录为特征，由Li等人报告，表明在光上阶段的唤醒时间显著增加，快速眼动（REM）睡眠减少，总睡眠和唤醒时间（在24小时内）在4周以上的建模24后不受影响。目前，总睡眠或部分睡眠剥夺是最常用的睡眠操纵模型。完全睡眠不足通常通过持续温和的处理或使动物接触新事物，或者通过连续旋转一个酒吧或跑步机25，26，27，28，29进行。由于道德原因，完全睡眠不足通常短于24小时。最常用的部分睡眠剥夺模型是水平台方法，它主要消融REM睡眠30，31，32。其他方法使用跑步机或酒吧，沿着笼子的底部扫，可能会诱发睡眠碎片时，设置在固定间隔33，34，35，36，37，38。值得注意的是，SF中断睡眠，断断续续地导致所有睡眠阶段的唤醒24。应用轨道转子的CSF模型的一个突出优点是，它可以连续运行几个月自动由机器控制，避免每天频繁处理劳动，除了定期监测。此外，该仪器将允许在军警干预下同时对多个老鼠笼子进行建模。在整个建模过程中，老鼠被安置在家庭笼子里，有通常的床上用品和筑巢材料，而其他一些方法则需要暴露在多样化的环境和不可避免的压力中。
睡眠分裂以前的特点是睡眠操纵方法，它模仿睡眠阶段频繁的觉醒和唤醒阶段的大量睡眠反弹。在一些文献中，CSF被认为是OSA39，40的动物模型。在这项研究中，选择的唤醒频率为每小时30次的原理是基于对中度至重度睡眠呼吸暂停患者的唤醒指数的观察。据观察，4周的睡眠分裂显著增加了超胶囊唤醒延迟和触觉唤醒阈值，这至少可以持续2周后恢复24。这种表型的解释是揭示c-fos活化减少诺拉迪纳吉奇，奥里辛热，组胺，和胆碱唤醒活性神经元，以回应高气喘，以及减少的儿茶酚胺和奥里辛ergic投影到结晶皮层24。然而，有必要指出，OSA最重要的特点是气道阻塞引起的缺氧，导致睡眠中断41，42。睡眠障碍和重复性缺氧在OSA发病机制中相互相互作用。因此，单靠睡眠分裂可能无法充分证明小鼠OSA的所有关键特征。
在此，我们提出了一个标准化的协议，以模拟年轻野生型小鼠的慢性睡眠分裂。在CSF治疗后，由莫里斯水迷宫、新奇物体识别、开放领域测试和强迫游泳测试评估了认知缺陷、焦虑和抑郁样行为。值得注意的是，这个模型应该作为一个整体，产生表型的调节不良的睡眠模式，认知缺陷，和焦虑般的行为。目前的模型可能适用于以下目的，但不限于以下目的：1） 进一步研究在没有遗传倾向的幼鼠慢性睡眠障碍引起的功能或分子发病机制， 2） 确定睡眠障碍引起的神经退化的直接途径，3） 探索改善慢性睡眠障碍诱发的表型的治疗方法，4） 研究野生型小鼠在慢性睡眠障碍下的内在保护/补偿机制，5） 应用于研究睡眠唤醒调节和状态过渡机制。

<table>
	<tbody>
		<tr>
			<td>Any-maze behavior tracking system</td>
			<td>Stoelting,Inc,USA</td>
			<td>-</td>
			<td>A video-tracking system which was used to record the behavior track of mice.</td>
		</tr>
		<tr>
			<td>C57BL/6J mice</td>
			<td>Hubei Research Center for Laboratory Animals, Hubei, China.</td>
			<td>-</td>
			<td>healthy male C57BL/6J mice aged 10-12 weeks were purchased from Hubei Research Center for Laboratory Animals</td>
		</tr>
		<tr>
			<td>Graphpad Prism 6.0 Software</td>
			<td>Graphpad Software,Inc.USA</td>
			<td>-</td>
			<td>Graphpad Prism 6.0 software was used to draw statistical graphs.</td>
		</tr>
		<tr>
			<td>Morris water maze system</td>
			<td>Shanghai XinRuan Information Technology Co.,Ltd,China</td>
			<td>XR-XM101</td>
			<td>The system was used to perform Morris water maze test</td>
		</tr>
		<tr>
			<td>Orbial rotor</td>
			<td>Shanghai ShiPing Laboratory Equipment Co.,Ltd,China</td>
			<td>SPH-331</td>
			<td>The orbital rotor was used to establish the chronic sleep fragmentation model</td>
		</tr>
		<tr>
			<td>Solid state timer</td>
			<td>OMRON Corporation, Kyoto, Japan</td>
			<td>H3CR-F8-300</td>
			<td>The solid state time was used to control the frequency and time of the rotor running</td>
		</tr>
		<tr>
			<td>Wooden Lusterless Tank</td>
			<td>-</td>
			<td>-</td>
			<td>length 30 cm, width 28 cm, height 35 cm The tank was used to perform open field test and novel object recognition test</td>
		</tr>
	</tbody>
</table>

a chronic sleep fragmentation model using vibrating orbital rotor to induce cognitive deficit and anxiety-like behavior in young wild-type mice

睡眠障碍通常作为慢性疾病或投诉事件在人群中很常见。慢性睡眠障碍建议与疾病的发病机制密切相关，特别是神经退行性疾病。我们最近发现，2个月的睡眠分裂引发了老年痴呆症（AD）样的行为和病理变化的年轻野生型小鼠。在这里，我们提出了一个标准化的协议，以实现慢性睡眠分裂（CSF）。简言之，CSF 是由轨道转子在 110 rpm 时振动引起的，在光打开阶段（上午 8：00-晚上 8：00）连续运行长达 2 个月，重复周期为 10 s 开，110 s-off。空间学习和记忆的损伤，焦虑样，但不是抑郁症样的行为，在小鼠作为CSF建模的后果，被评估与莫里斯水迷宫（MWM），新颖的对象识别（NOR），开放领域测试（OFT）和强迫游泳测试（FST）。与其他睡眠操作相比，此协议最大限度地减少了处理工作，并最大限度地提高了建模效率。它在年轻的野生型小鼠中产生稳定的表型，并有可能产生用于各种研究目的。

所有有代表性的结果和数字均由我们最近出版的第21期转载。原期刊允许重复使用这些数字。
整个实验设计按时间顺序进行说明，表示CSF建模的时间、MWM、NOR、OFT和FST（图1A）的行为测试。我们每周从CSF和对照组获得老鼠的重量，以监测它们在建模过程中的一般情况。在建模过程中，两组小鼠的体重增加没有明显差异（?...

Watch this Scientific Journal Video about A Chronic Sleep Fragmentation Model using Vibrating Orbital Rotor to Induce Cognitive Deficit and Anxiety-Like Behavior in Young Wild-Type Mice at JoVE.com

A Chronic Sleep Fragmentation Model using Vibrating Orbital Rotor to Induce Cognitive Deficit and Anxiety-Like Behavior in Young Wild-Type Mice

这里介绍的是一个由电控轨道转子实现的慢性睡眠碎片（CSF）模型的协议，它可以诱导年轻野生型小鼠的认知缺陷和类似焦虑的行为。该模型可用于探索慢性睡眠障碍和相关疾病的发病机制。

使用振动轨道转子诱导年轻野生型小鼠认知缺陷和焦虑样行为的慢性睡眠碎片模型

Sleep disturbance is generally common in populations as a chronic disease or a complained event. Chronic sleep disturbance is proposed to be closely ...

a-chronic-sleep-fragmentation-model-using-vibrating-orbital-rotor-to

Research

JoVE Journal

Behavior

5.2K 次观看.  Huazhong University of Science and Technology, Wuhan 430000,  China. 这里介绍的是一个由电控轨道转子实现的慢性睡眠碎片（CSF）模型的协议，它可以诱导年轻野生型小鼠的认知缺陷和类似焦虑的行为。该模型可用于探索慢性睡眠障碍和相关疾病的发病机制。

视频: A Chronic Sleep Fragmentation Model using Vibrating Orbital Rotor to Induce Cognitive Deficit and Anxiety-Like Behavior in Young Wild-Type Mice

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

The Crossmodal Congruency Task as a Means to Obtain an Objective Behavioral Measure in the Rubber Hand Illusion Paradigm

The rubber hand illusion (RHI) is a popular experimental paradigm. Participants view touch on an artificial rubber hand while the participants' own hidden hand is touched. If the viewed and felt touches are given at the same time then this is sufficient to induce the compelling experience that the rubber hand is one's own hand. The RHI can be used to investigate exactly how the brain constructs distinct body representations for one's own body. Such representations are crucial for successful interactions with the external world. To obtain a subjective measure of the RHI, researchers typically ask participants to rate statements such as "I felt as if the rubber hand were my hand". Here we demonstrate how the crossmodal congruency task can be used to obtain an objective behavioral measure within this paradigm.
The variant of the crossmodal congruency task we employ involves the presentation of tactile targets and visual distractors. Targets and distractors are spatially congruent (i.e. same finger) on some trials and incongruent (i.e. different finger) on others. The difference in performance between incongruent and congruent trials - the crossmodal congruency effect (CCE) - indexes multisensory interactions. Importantly, the CCE is modulated both by viewing a hand as well as the synchrony of viewed and felt touch which are both crucial factors for the RHI.
The use of the crossmodal congruency task within the RHI paradigm has several advantages. It is a simple behavioral measure which can be repeated many times and which can be obtained during the illusion while participants view the artificial hand. Furthermore, this measure is not susceptible to observer and experimenter biases. The combination of the RHI paradigm with the crossmodal congruency task allows in particular for the investigation of multisensory processes which are critical for modulations of body representations as in the RHI.

We demonstrate how an objective measure can be employed in the widely employed rubber hand illusion paradigm. This measure is obtained by modifying the well-established crossmodal congruency task. This task allows the investigation of multisensory processes which are critical for modulations of body representations as in the rubber hand illusion.

一致性的任务作为一种手段来获取目标行为的措施中的橡胶手错觉范式Crossmodal

The rubber hand illusion (RHI) is a popular experimental paradigm. Participants view touch on an artificial rubber hand while the participants' own hidden ...

Eye Tracking, Cortisol, and a Sleep vs. Wake Consolidation Delay: Combining Methods to Uncover an Interactive Effect of Sleep and Cortisol on Memory

Although rises in cortisol can benefit memory consolidation, as can sleep soon after encoding, there is currently a paucity of literature as to how these two factors may interact to influence consolidation. Here we present a protocol to examine the interactive influence of cortisol and sleep on memory consolidation, by combining three methods: eye tracking, salivary cortisol analysis, and behavioral memory testing across sleep and wake delays. To assess resting cortisol levels, participants gave a saliva sample before viewing negative and neutral objects within scenes. To measure overt attention, participants&#8217; eye gaze was tracked during encoding. To manipulate whether sleep occurred during the consolidation window, participants either encoded scenes in the evening, slept overnight, and took a recognition test the next morning, or encoded scenes in the morning and remained awake during a comparably long retention interval. Additional control groups were tested after a 20&#160;min&#160;delay in the morning or evening, to control for time-of-day effects. Together, results showed that there is a direct relation between resting cortisol at encoding and subsequent memory, only following a period of sleep. Through eye tracking, it was further determined that for negative stimuli, this beneficial effect of cortisol on subsequent memory may be due to cortisol strengthening the relation between where participants look during encoding and what they are later able to remember. Overall, results obtained by a combination of these methods uncovered an interactive effect of sleep and cortisol on memory consolidation.

We present a protocol used to discover an interactive effect between sleep and cortisol on memory consolidation, particularly for negative arousing images. Specifically, the experimental design utilizes eye tracking, salivary cortisol analysis, and behavioral memory testing &#8211; methods that can be used with both healthy and clinical participants.

眼睛跟踪，皮质醇和睡眠与唤醒延迟合并：合并方法来揭开睡眠和皮质醇的交互式影响记忆

Although rises in cortisol can benefit memory consolidation, as can sleep soon after encoding, there is currently a paucity of literature as to how these ...

Uncovering Beat Deafness: Detecting Rhythm Disorders with Synchronized Finger Tapping and Perceptual Timing Tasks

A set of behavioral tasks for assessing perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) is presented here with the goal of uncovering rhythm disorders, such as beat deafness. Beat deafness is characterized by poor performance in perceiving durations in auditory rhythmic patterns or poor synchronization of movement with auditory rhythms (e.g., with musical beats). These tasks include the synchronization of finger tapping to the beat of simple and complex auditory stimuli and the detection of rhythmic irregularities (anisochrony detection task) embedded in the same stimuli. These tests, which are easy to administer, include an assessment of both perceptual and sensorimotor timing abilities under different conditions (e.g., beat rates and types of auditory material) and are based on the same auditory stimuli, ranging from a simple metronome to a complex musical excerpt. The analysis of synchronized tapping data is performed with circular statistics, which provide reliable measures of synchronization accuracy (e.g., the difference between the timing of the taps and the timing of the pacing stimuli) and consistency. Circular statistics on tapping data are particularly well-suited for detecting individual differences in the general population. Synchronized tapping and anisochrony detection are sensitive measures for identifying profiles of rhythm disorders and have been used with success to uncover cases of poor synchronization with spared perceptual timing. This systematic assessment of perceptual and sensorimotor timing can be extended to populations of patients with brain damage, neurodegenerative diseases (e.g., Parkinson&#8217;s disease), and developmental disorders (e.g., Attention Deficit Hyperactivity Disorder).

Behavioral tasks that allow for the assessment of perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) are presented. Synchronization of finger tapping to the beat of an auditory stimuli and detecting rhythmic irregularities provides a means of uncovering rhythm disorders.

揭开节拍耳聋：检测节律紊乱与同步手指敲击和感知定时任务

A set of behavioral tasks for assessing perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) is presented here ...

Use of the Open Field Maze to Measure Locomotor and Anxiety-like Behavior in Mice

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of the most commonly used platforms to measure behaviors in animal models. It is a fast and relatively easy test that provides a variety of behavioral information ranging from general ambulatory ability to data regarding the emotionality of the subject animal. As it relates to rodent models, the procedure allows the study of different strains of mice or rats both laboratory bred and wild-captured. The technique also readily lends itself to the investigation of different pharmacological compounds for anxiolytic or anxiogenic effects. Here, a protocol for use of the open field maze to describe mouse behaviors is detailed and a simple analysis of general locomotor ability and anxiety-related emotional behaviors between two strains of C57BL/6 mice is performed. Briefly, using the described protocol we show Wild Type mice exhibited significantly less anxiety related behaviors than did age-matched Knock Out mice while both strains exhibited similar ambulatory ability.

A protocol is provided to use an Open Field Maze to access general locomotor activity, anxiety and emotionality in a laboratory mouse model.

使用开放式现场迷宫来衡量小鼠的运动能力和焦虑样行为

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of ...

Introducing Clicker Training as a Cognitive Enrichment for Laboratory Mice

Establishing new refinement strategies in laboratory animal science is a central goal in fulfilling the requirements of Directive 2010/63/EU. Previous research determined a profound impact of gentle handling protocols on the well-being of laboratory mice. By introducing clicker training to the keeping of mice, not only do we promote the amicable treatment of mice, but we also enable them to experience cognitive enrichment. Clicker training is a form of positive reinforcement training using a conditioned secondary reinforcer, the "click" sound of a clicker, which serves as a time bridge between the strengthened behavior and an upcoming reward. The effective implementation of the clicker training protocol with a cohort of 12 BALB/c inbred mice of each sex proved to be uncomplicated. The mice learned rather quickly when challenged with tasks of the clicker training protocol, and almost all trained mice overcame the challenges they were given (100% of female mice and 83% of male mice). This study has identified that clicker training for mice strongly correlates with reduced fear in the mice during human-mice interactions, as shown by reduced anxiety-related behaviors (e.g., defecation, vocalization, and urination) and fewer depression-like behaviors (e.g., floating). By developing a reliable protocol that can be easily integrated into the daily routine of the keeping of laboratory mice, the lifetime experience of welfare in the mice can be improved substantially.

The development of new refinement strategies for laboratory mice is a challenging task that contributes towards fulfilling the 3R principle. This protocol introduces clicker training as a cognitive enrichment program for laboratory mice.

引进响片训练为实验室老鼠认知富集

Establishing new refinement strategies in laboratory animal science is a central goal in fulfilling the requirements of Directive 2010/63/EU. Previous ...

Chronic Sleep Deprivation in Mouse Pups by Means of Gentle Handling

Sleep is critical for proper development and neural plasticity. Moreover, abnormal sleep patterns are characteristic of many neurodevelopmental disorders. Studying how chronic sleep restriction during development can affect adult behavior may add to our understanding of the emergence of behavioral symptoms of neurodevelopmental disorders. While there are many methods that can be used to restrict sleep in rodents including forced locomotion, constant disruption, presentation of an aversive stimulus, or electric shock, many of these methods are very stressful and cannot be used in neonatal mice. Here, we describe gentle handling, a sleep deprivation technique that can be used chronically throughout development and into adulthood to achieve sleep restriction. Gentle handling involves close observation of the mice throughout the sleep deprivation period and requires the researcher to gently prod the animals whenever they are inactive or display behaviors associated with sleep. Coupled with EEG recordings, gentle handling could be used to selectively disrupt a specific phase of sleep such as rapid eye movement (REM) sleep. The technique of gentle handling is a powerful tool for the study of the effects of chronic sleep restriction even in neonatal mice that circumvents many of the more stressful procedures used for sleep deprivation.

We describe a sleep deprivation technique known as gentle handling where investigators gently prod mice any time sleep behavior is observed. This method is a powerful tool that allows researchers to study the effects that chronic sleep restriction throughout development can have on future brain physiology and behavior.

小鼠幼犬慢性睡眠剥夺的温和处理方法

Sleep is critical for proper development and neural plasticity. Moreover, abnormal sleep patterns are characteristic of many neurodevelopmental disorders. ...

Novel Object Recognition and Object Location Behavioral Testing in Mice on a Budget

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various physiological or pathological changes. The object location task (OLT) and the novel object recognition task (NORT) are two effective behavioral tasks commonly used to reveal the function and relative health of specific brain regions involved in memory. While both of these tests exploit the inherent preference of mice for the novelty to reveal memory for previously encountered objects, the OLT primarily evaluates spatial learning, which relies heavily on hippocampal activity. The NORT, in contrast, evaluates non-spatial learning of object identity, which relies on multiple brain regions. Both tasks require an open-field-testing arena, objects with equivalent intrinsic value to mice, appropriate environmental cues, and video recording equipment and the software. Commercially available systems, while convenient, can be costly. This manuscript details a simple, cost-effective method for building the arenas and setting up the equipment necessary to perform the OLT and NORT. Furthermore, the manuscript describes an efficient testing protocol that incorporates both OLT and NORT and provides typical methods for data acquisition and analysis, as well as representative results. Successful completion of these tests can provide valuable insight into the memory function of various mouse model systems and appraise the underlying neural regions that support these functions.

Here we provide a protocol which includes comprehensive instructions for the economical establishment of murine object location and novel object recognition behavioral testing, including the design, cost, and construction of required equipment as well as execution of behavioral testing, data collection, and analysis.

一种新的小鼠目标识别与目标定位行为测试

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various ...

A Chronic Immobilization Stress Protocol for Inducing Depression-Like Behavior in Mice

Depression is not yet fully understood, but various causative factors have been reported. Recently, the prevalence of depression has increased. However, therapeutic treatments for depression or research on depression is scarce. Thus, in the present paper, we propose a mouse model of depression induced by movement restriction. Chronic mild stress (CMS) is a well-known technique to induce depressive-like behavior. However, it necessitates a complex procedure consisting of a combination of various mild stresses. In contrast, chronic immobilization stress (CIS) is a readily accessible chronic stress model, modified from a restraint model that induces depressive behavior by restricting movement using a restrainer for a certain period. To evaluate the depressive-like behaviors, the sucrose preference test (SPT), the tail suspension test (TST), and the ELISA assay to measure stress marker corticosterone levels are combined in the present experiment. The described protocols illustrate the induction of CIS and evaluation of the changes in behavior and physiological factors for the validation of depression.

This article provides a simplified and standardized protocol for induction of depressive-like behavior in chronically immobilized mice by using a restrainer. In addition, behavior and physiological techniques to verify induction of depression are explained.

诱导小鼠抑郁行为的慢性固定性压力协议

Depression is not yet fully understood, but various causative factors have been reported. Recently, the prevalence of depression has increased. However, ...

Nest Building Behavior as an Early Indicator of Behavioral Deficits in Mice

Nest building is an innate behavior in male and female rodents, even when raised in laboratory settings. As such, many researchers provide rodents synthetic and/or natural materials (such as twine, tissue, cotton, paper, and hay) as a gauge of their overall well-being and as an ancillary assessment to predict the possible decline in cognition. Typically, changes in nesting behaviors, such as failure to create a nest, indicate a change in health or welfare. In addition, nesting behavior is sensitive to many environmental and physiological challenges, as well as many genetic mutations underlying pathological disease states. The following protocol describes a nesting behavior paradigm that explores the usage of four types of nesting material. In addition, the protocol utilizes intraclass correlations to demonstrate that inter-rater reliability is higher when nests are constructed out of shredded paper compared to other common nesting materials such as cotton squares, paper twists, and soft cob bedding. The chosen methodology and statistical considerations (i.e., intraclass correlation) for this assay may be of interest for those conducting experiments assessing the quality of living of mice.&#160;

Here, we present a protocol to quantify nest building behavior in mice, which is known to be impaired in several neurological disorders and diseases. This protocol examines the utility of four materials and offers the opportunity to quantify the rater agreement in scoring, improving the validity and reliability of the assay.