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

Applying Stereotactic Injection Technique to Study Genetic Effects on Animal Behaviors

Stereotactic injection is a useful technique to deliver high titer lentiviruses to targeted brain areas in mice. Lentiviruses can either overexpress or knockdown gene expression in a relatively focused region without significant damage to the brain tissue. After recovery, the injected mouse can be tested on various behavioral tasks such as the Open Field Test (OFT) and the Forced Swim Test (FST). The OFT is designed to assess locomotion and the anxious phenotype in mice by measuring the amount of time that a mouse spends in the center of a novel open field. A more anxious mouse will spend significantly less time in the center of the novel field compared to controls. The FST assesses the anti-depressive phenotype by quantifying the amount of time that mice spend immobile when placed into a bucket of water. A mouse with an anti-depressive phenotype will spend significantly less time immobile compared to control animals. The goal of this protocol is to use the stereotactic injection of a lentivirus in conjunction with behavioral tests to assess how genetic factors modulate animal behaviors.

Stereotactic injection of lentiviruses expressing cDNAs or shRNAs can modulate gene expression in specific brain areas of mice. Here, we present a protocol to combine stereotactic injections with behavioral tasks, such as the Open Field Test (OFT) and the Forced Swim Test (FST).

应用立体定向注射技术在研究动物行为的遗传效应

Stereotactic injection is a useful technique to deliver high titer lentiviruses to targeted brain areas in mice. Lentiviruses can either overexpress or ...

科研

行为学

Noninvasive, High-throughput Determination of Sleep Duration in Rodents

Traditionally, sleep is monitored by an electroencephalogram (EEG). EEG studies in rodents require surgical implantation of the electrodes followed by a long recovery period. To perform an EEG recording, the animal is connected to a receiver, creating an unnatural tether to the head-mount. EEG monitoring is time consuming, carries risk to the animal, and is not a completely natural setting for the measurement of sleep. Alternative methods to detect sleep, particularly in a high-throughput fashion, would greatly advance the field of sleep research. Here, we describe a validated method for detecting sleep via activity-based home-cage monitoring. Previous studies have shown that sleep assessed via this method has a high degree of agreement with sleep defined by traditional EEG-based measures. Whereas this method is validated for total sleep time, it is important to note that sleep bout duration should be assessed by an EEG which has better temporal resolution. The EEG can also differentiate rapid eye movement (REM) and non-REM sleep, giving more detail about the exact nature of sleep. Nevertheless, activity-based sleep determination can be used to analyze multiple days of undisturbed sleep and to assess sleep as a response to an acute event (like stress). Here, we show the power of this system to detect the response of mice to daily intraperitoneal injections.

We describe a high-throughput method of measuring sleep by means of activity-based home-cage monitoring. This method offers advantages over traditional EEG-based methods. It is well validated for the determination of total sleep duration and can be a powerful tool to monitor sleep in rodent models of human disease.

无创高通量测定啮齿类动物睡眠时间

Traditionally, sleep is monitored by an electroencephalogram (EEG). EEG studies in rodents require surgical implantation of the electrodes followed by a ...

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. ...

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, ...

Quantifying Infra-slow Dynamics of Spectral Power and Heart Rate in Sleeping Mice

Three vigilance states dominate mammalian life: wakefulness, non-rapid eye movement (non-REM) sleep, and REM sleep. As more neural correlates of behavior are identified in freely moving animals, this three-fold subdivision becomes too simplistic. During wakefulness, ensembles of global and local cortical activities, together with peripheral parameters such as pupillary diameter and sympathovagal balance, define various degrees of arousal. It remains unclear the extent to which sleep also forms a continuum of brain states-within which the degree of resilience to sensory stimuli and arousability, and perhaps other sleep functions, vary gradually-and how peripheral physiological states co-vary. Research advancing the methods to monitor multiple parameters during sleep, as well as attributing to constellations of these functional attributes, is central to refining our understanding of sleep as a multifunctional process during which many beneficial effects must be executed. Identifying novel parameters characterizing sleep states will open opportunities for novel diagnostic avenues in sleep disorders.
We present a procedure to describe dynamic variations of mouse non-REM sleep states via the combined monitoring and analysis of electroencephalogram (EEG)/electrocorticogram (ECoG), electromyogram (EMG), and electrocardiogram (ECG) signals using standard polysomnographic recording techniques. Using this approach, we found that mouse non-REM sleep is organized into cycles of coordinated neural and cardiac oscillations that generate successive 25-s intervals of high and low fragility to external stimuli. Therefore, central and autonomic nervous systems are coordinated to form behaviorally distinct sleep states during consolidated non-REM sleep. We present surgical manipulations for polysomnographic (i.e., EEG/EMG combined with ECG) monitoring to track these cycles in the freely sleeping mouse, the analysis to quantify their dynamics, and the acoustic stimulation protocols to assess their role in the likelihood of waking up. Our approach has already been extended to human sleep and promises to unravel common organizing principles of non-REM sleep states in mammals.

Here, we present experimental and analytical procedures to describe the temporal dynamics of the neural and cardiac variables of non-REM sleep in mice, which modulate sleep responsiveness to acoustic stimuli.

量化睡眠小鼠的光谱功率和心率的慢速动力学

Three vigilance states dominate mammalian life: wakefulness, non-rapid eye movement (non-REM) sleep, and REM sleep. As more neural correlates of behavior ...

神经科学

A Rat Model of Central Fatigue Using a Modified Multiple Platform Method

In this article, we introduced a rat model of central fatigue using the modified multiple platform method (MMPM). The Multiple Platform box was designed as a water tank with narrow platforms on the bottom. The model rats were put into the tank and stood on the platforms for 14 h (18:00 - 8:00) per day for a consecutive 21 days, with a blank control group set for contrast. At the end of modeling, rats in the model group showed an obvious fatigued appearance. To assess the model, we performed several behavioral tests, including the open field test (OFT), the elevated plus maze (EPM) test, and the exhaustive swimming (ES) test. The results showed that anxiety, spatial cognition impairment, poor muscle performance, and declined voluntary activity presented in model rats confirm the diagnosis of&#160;central fatigue. Changes of the central neurotransmitters also verified the result. In conclusion, the model successfully simulated central fatigue, and future study with the model may help reveal the pathological mechanism of the disease.

Here, we present a protocol to introduce a rat model of central fatigue using the modified multiple platform method (MMPM).

基于改进多平台法的大鼠中心疲劳模型

In this article, we introduced a rat model of central fatigue using the modified multiple platform method (MMPM). The Multiple Platform box was designed ...

Optogenetic Manipulation of Neural Circuits During Monitoring Sleep/wakefulness States in Mice

In recent years, optogenetics has been widely used in many fields of neuroscientific research. In many cases, an opsin, such as channel rhodopsin 2 (ChR2), is expressed by a virus vector in a particular type of neuronal cells in various Cre-driver mice. Activation of these opsins is triggered by application of light pulses which are delivered by laser or LED through optic cables, and the effect of activation is observed with very high time resolution. Experimenters are able to acutely stimulate neurons while monitoring behavior or another physiological outcome in mice. Optogenetics can enable useful strategies to evaluate function of neuronal circuits in the regulation of sleep/wakefulness states in mice. Here we describe a technique for examining the effect of optogenetic manipulation of neurons with a specific chemical identity during electroencephalogram (EEG) and electromyogram (EMG) monitoring to evaluate the sleep stage of mice. As an example, we describe manipulation of GABAergic neurons in the bed nucleus of the stria terminalis (BNST). Acute optogenetic excitation of these neurons triggers a rapid transition to wakefulness when applied during NREM sleep. Optogenetic manipulation along with EEG/EMG recording can be applied to decipher the neuronal circuits that regulate sleep/wakefulness states.

Here, we describe methods of optogenetic manipulation of particular types of neurons during monitoring of sleep/wakefulness states in mice, presenting our recent work on the bed nucleus of the stria terminalis as an example.

监测小鼠睡眠/觉醒状态期间神经回路的光遗传学操作

In recent years, optogenetics has been widely used in many fields of neuroscientific research. In many cases, an opsin, such as channel rhodopsin 2 ...

Electrocorticographic Recording of Cerebral Cortex Areas Manipulated Using an Adeno-Associated Virus Targeting Cofilin in Mice

The use of electrocorticographic (ECoG) recordings in rodents is relevant to sleep research and to the study of a wide range of neurological conditions. Adeno-associated viruses (AAVs) are increasingly used to improve understanding of brain circuits and their functions. The AAV-mediated manipulation of specific cell populations and/or of precise molecular components has been tremendously useful to identify new sleep regulatory circuits/molecules and key proteins contributing to the adverse effects of sleep loss. For instance, inhibiting activity of the filamentous actin-severing protein&#160;cofilin&#160;using AAV prevents sleep deprivation-induced memory impairment. Here, a protocol is&#160;described that combines the manipulation of cofilin function in a cerebral cortex area with the recording of ECoG activity to examine whether cortical cofilin modulates the wakefulness and sleep ECoG signals. AAV injection is performed during the same surgical procedure as the implantation of ECoG and electromyographic (EMG) electrodes in adult male and female mice. Mice are anesthetized, and their heads are shaved. After skin cleaning and incision, stereotaxic coordinates of the motor cortex are determined, and the skull is pierced at this location. A cannula prefilled with an AAV expressing cofilinS3D, an inactive form of cofilin, is slowly positioned in the cortical tissue. After AAV infusion, gold-covered screws (ECoG electrodes) are screwed through the skull and cemented to the skull with gold wires inserted in the neck muscles (EMG electrodes). The animals are allowed three weeks to recover and to ensure sufficient expression of cofilinS3D. The infected area and cell type are verified using immunohistochemistry, and the ECoG is analyzed using visual identification of vigilance states and spectral analysis. In summary, this combined methodological approach allows the investigation of the precise contribution of molecular components regulating neuronal morphology and connectivity to the regulation of synchronized cerebral cortex activity during wakefulness and sleep.

This article describes a protocol for the manipulation of molecular targets in the cerebral cortex using adeno-associated viruses and for monitoring the effects of this manipulation during wakefulness and sleep using electrocorticographic recordings.

使用腺相关病毒在小鼠中瞄准科菲林操纵的大脑皮层区域的电皮质记录

The use of electrocorticographic (ECoG) recordings in rodents is relevant to sleep research and to the study of a wide range of neurological conditions. ...

Social Isolation Model: A Noninvasive Rodent Model of Stress and Anxiety

Anxiety disorders are one of the leading causes of disability in the United States (US). Current treatments are not always effective and less than 50% of patients achieve full remission. A critical step in developing a novel anxiolytic is to develop and utilize an animal model, such as mice, to study pathological changes and test drug target(s), efficacy, and safety. Current approaches include genetic manipulation, chronic administration of anxiety-inducing molecules, or the administration of environmental stress. These methods, however, may not realistically reflect anxiety induced throughout daily life. This protocol describes a novel anxiety model, which mimics the intentional or unintentional patterns of social isolation in modern life. The social isolation-induced anxiety model minimizes perceived distractions and invasiveness and utilizes wild type C57BL/6 mice. In this protocol, 6- to 8-week-old mice (male and female) are singly housed in opaque cages to visually block the external environment, such as neighboring mice, for 4 weeks. No environmental enrichments (such as toys) are provided, bedding material is reduced by 50%, any treatment of drug is administered as an agar form, and the exposure/handling of the mice is minimized. Socially isolated mice generated using this protocol exhibit greater anxiety-like behavior, aggression,&#160;as well as decreased cognition.

Presented here is a social isolation (SI)-induced anxiety mouse model that utilizes wild type C56BL/6J mice to induce stress and anxiety-like behavior with minimal handling and no invasive procedures. This model reflects modern life patterns of social isolation and is ideal for studying anxiety and related disorders.

社会隔离模型:压力和焦虑的非侵入性啮齿动物模型

Anxiety disorders are one of the leading causes of disability in the United States (US). Current treatments are not always effective and less than 50% of ...

Establishing a Device for Sleep Deprivation in Mice

Circadian rhythm disruption refers to the desynchronization between the external environment or behavior and the endogenous molecular clock, which significantly impairs health. Sleep deprivation is one of the most common causes of circadian rhythm disruption. Various modalities (e.g., platforms on the water, gentle handling, sliding bar chambers, rotating drums, orbital shakers, etc.) have been reported for inducing sleep deprivation in mice to investigate its effects on health. The current study introduces an alternative method for sleep deprivation in mice. An automated rocker platform-based device was designed that is cost-effective and efficiently disrupts sleep in group-housed mice at adjustable time intervals. This device induces characteristic changes of sleep deprivation with minimal stress response. Consequently, this method may prove useful for investigators interested in studying the effects and underlying mechanisms of sleep deprivation on the pathogenesis of multiple diseases. Moreover, it offers a cost-effective solution, particularly when multiple sleep deprivation devices are required to run in parallel.

The present protocol outlines a method for setting up a cost-effective rocker platform-based device used for inducing sleep deprivation in mice. This device has proven to be effective in causing disruptions in electroencephalogram (EEG)-evidenced sleep patterns, as well as inducing metabolic and molecular changes associated with sleep deprivation.