作者希望感谢 NIH 研究员编辑委员会的编辑协助。这项研究由镍氢 (MH00889) 的壁内研究计划资助。RMS 也得到了 FRAXA 博士后奖学金的支持。

所有程序均经国家心理健康动物护理和使用委员会批准, 并按照国家卫生研究所关于动物的照料和使用指导方针进行。
1. 设置睡眠检测单元
<ol><li>购买所需的单位和软件数量。</li>
	<li>按照说明设置监控系统。<ol>
<li>对准发射器对面的探测器。确保红外光束朝内, 并在同一高度上对齐。</li>
		<li>使用所提供的螺钉将探测器和发射器放置在金属支架所需的高度上。这种高度应调整, 使笼的床上用品低于红外光束的水平, 但光束是在适当的高度, 以检测动物的活动。这将创建一个27厘米 x 32 厘米的内部区域 (10.625 x 12.75 英寸)。<ol>
<li>如果需要更多的螺丝, 购买更多的硬件商店 (6-32 ¼在潘头螺钉)。</li>
		</ol>
</li>
	</ol>
</li>
	<li>将每个探测器与每个发射器连接起来。将发射器连接到与接收器相连的所提供的中枢。对 x 和 y 平面都执行此操作。对所有设置重复此步骤。</li>
	<li>使用提供的 USB 集线器将接收器连接到计算机。</li>
</ol>2. 软件设置
<ol><li>安装分析软件并选择提供的硬件配置文件作为默认值。硬件配置文件只应由公司更改。</li>
	<li>单击文件 |打开实验配置。打开默认的实验配置。</li>
	<li>单击实验 |属性。</li>
	<li>单击扫描选项卡. 将扫描速率更改为十年代。</li>
	<li>单击活动选项卡. 将活动采样率更改为十年代。</li>
	<li>单击文件 |将实验保存为以保存这些配置设置以便将来进行实验。</li>
</ol>3. 动物设置
<ol><li>单独安置老鼠在干净的笼子里是 31 cm (12.25 英寸) 长和 16.5 cm (6.5 英寸) 宽。为了防止老鼠建立床上用品和阻碍梁, 使用床上用品在深度3毫米。不要为老鼠提供额外的筑巢材料。</li>
	<li>为老鼠提供食物和水的途径ad 随意通过一个电线馈线, 在笼子的顶部, 从梁的方式。如有必要, 在整个研究期间, 每3-5 天更换一只笼子, 将食物重新灌装并更换水瓶。</li>
	<li>将鼠标笼对准红外光束设置, 确保其位于光束的中间, 以充分覆盖。</li>
	<li>确保房间的光暗循环被设置为反射小鼠正常居住条件的光暗循环, 或者根据实验的需要进行改变。</li>
</ol>4. 药物制剂和注射剂
<ol><li>获得无菌水和无菌环糊精。</li>
	<li>对于所有注射, 使用1毫升注射器与 25.5-30.5 克皮下注射针。每只老鼠都应该使用一种新的无菌针头和注射器。</li>
	<li>为了缩短注射时间, 请提前组装所有注射器。</li>
	<li>用于环糊精注射: 将3克环糊精添加到10毫升水中, 制备环糊精溶液, 制作30% 环糊精溶液。</li>
	<li>通过 IP 注射管理0.3 毫升的生理盐水或环糊精。</li>
</ol>5. 记录睡眠的软件
<ol><li>打开默认硬件配置文件。</li>
	<li>单击文件 |打开实验配置。打开所需的或默认的实验配置。</li>
	<li>单击实验 |安装程序。<ol>
<li>指定保存数据文件的位置以及保存备份文件的位置。该软件要求数据文件存储在特定程序文件夹下的程序文件中。 注意: 由于某些计算机将程序文件视为敏感的, 因此在实验之后可能无法访问数据文件。因此, 建议将备份文件保存到本地计算机上的另一个不敏感区域。</li>
		<li>指定动物 ID 到每个睡眠室并且输入动物的重量, 如果需要。如果实验中没有使用腔室, 则取消选中该框将停用该腔。输入标识信息后, 单击完成。</li>
	</ol>
</li>
	<li>单击文件 |将实验保存为以将当前的实验配置保存在所需的文件名下。</li>
	<li>单击实验 |运行或F5启动录制。等待一个十年代的时代, 以确保所有的房间都在回升活动。当动物被注射的时候, 很有可能这些动物会移动到足以被红外光束探测到。<ol>
<li>如果未检测到任何活动, 请尝试手动移动鼠标笼, 以确保光束拾取移动。如果仍未记录活动, 则继续故障排除或移至工作室。 注: 在注射完成后, 立即运行实验。</li>
	</ol>
</li>
	<li>用不透明的盖子遮住电脑屏幕, 挡住光线。覆盖活动指示器上的闪烁灯。</li>
	<li>运行实验24小时。 注: 由于需要的时间来执行注射, 它不会是一个完整的24小时。注射剂应在每天的同一时间进行。在注射过程中不应记录睡眠。</li>
	<li>第二天, 在注射时, 单击实验 |停止以结束录制。单击文件 |出口 |生成主题 CSVs收集每个鼠标的原始数据。 注意: 导出可能需要一些时间, 所以立即执行此操作, 然后在计算机处理时进行注射。</li>
	<li>单击文件 |出口 |休眠通过打开 raw 来导出每个实验的休眠文件。CDTA 安装了软件的任何计算机上的数据文件。<ol>
<li>在检测参数活动源下, 确保选中了 x 轴和 y 轴框。</li>
		<li>在休眠阈值世纪下, 确保选择了4个纪元。</li>
		<li>在休眠阈值活动阈值下, 确保选中了0计数。</li>
		<li>在光/暗循环下, 检查光照/暗循环的适当时间。</li>
		<li>在分析窗口下, 选择所需的分析时间。在注入研究的情况下, 将日期保留为默认值。将开始时间设置为 ExpSTART。将持续时间设置为 24:00, 24 h. 使用该软件的分析最多可以执行72小时。</li>
		<li>保存配置以节省导出数据的时间。</li>
		<li>单击更新。</li>
		<li>单击生成 CSV 文件并将文件保存到所需位置。</li>
	</ol>
</li>
</ol>6. 数据分析
<ol><li>对于每个录制会话, 打开休眠文件。若要分配主题 id, 请打开每个分庭的单个 CSV 文件以确定主题 id。记录所有动物的光和暗相的hh:mm:ss 睡眠的。<ol>
<li>检查单个主题 CSV 文件中是否存在表明记录失败的不一致。如果在一平面中检测到高光束计数, 但在另一平面上没有观测到任何计数, 则表示光束故障。 注: 光和暗相的 "% 睡眠" 计算在整个记录期间, 包括光和暗相。对于12小时的光/暗循环, 将这个数字乘以 2, 得到 12 h 光或暗相的 "% 睡眠"。鉴于注射研究并没有完全24小时, 计算出的 "% 睡眠" 是不准确的。要生成正确的 "% 睡眠", 将 "hh:mm:ss 睡眠" 除以 "实验运行时间: hh:mm:ss", 乘以 2, 然后乘以100以获得百分比。如果在光相中进行注射, 则只有光相需要以这种方式进行调整。</li>
	</ol>
</li>
	<li>由于不同的天和阶段正在进行比较, 分析每个阶段和每天的睡眠时间百分比。</li>
</ol>

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作者没有什么可透露的。

在这里, 我们提出了一种无创, 高通量的方法, 以确定睡眠时间的基础上的活动监测的家庭笼。这种评估总睡眠时间的方法已经验证了对脑电图研究3。基于活动的家庭网箱监测方法简单, 无创, 适用于大量动物的人口研究。它是有限的, 因为它不能提供睡眠的详细信息 (如睡眠时间和睡眠阶段)。
这种分析方法的缺陷相当容易检测。重要的是, 在研究中没有任何?...

睡眠有重要的功能, 恢复身体和大脑后的每日负担的觉醒1。已经表明睡眠在记忆保留和一般大脑可塑性方面起着作用1。EEG 是检测睡眠2的黄金标准。在啮齿目动物, 脑电图监测需要手术植入的电极贴在头部安装, 之后, 动物需要一段时间来恢复2。恢复后, 该动物附着在录音设备上, 并给予另一段适应期2。由于这些必要的恢复和适应的时期, EEG 是费时费力的, 不能合理地执行大规模。此外, 电极植入的手术过程对动物有内在的危险。最后, 对脑电研究中评分睡眠的数据分析也非常费力。一种替代的、非侵入性的、高通量的睡眠监测方法将极大地帮助啮齿动物睡眠研究。
一种基于活动的家庭网箱监测系统用于检测睡眠, 解决了脑电图研究的局限性。简单的前提是不活泼的动物很可能是沉睡的动物。它已经表明, 四十年代连续不活动 (作废在十年代世纪) 是一个可靠的测量睡眠以脑电图测量 (显示有88-94% 协议)3。家庭网箱监测系统可用于研究大群动物, 设置时间最少。我们已经表明, 动物需要大约一天的时间习惯到家庭笼监控系统中的单个外壳, 与脑电图研究2所需的几周的恢复相比,4 。此外, 一些设置还可以检测的生理参数, 如核心体温, 心率, 活动, 和喂养。温度和心率是由一个小发射机的植入决定的。这些参数可以提供有关鼠标的更多信息, 并可与睡眠记录并行使用, 进一步增加我们对睡眠的理解以及它是如何受到影响的。
虽然它是一个强大的工具, 但对于可以从基于活动的家庭笼监视中获取的数据类型有一些限制。脑电图研究可以区分 rem 和非 rem 睡眠, 这可能是一个重要的深入了解睡眠结构。基于活动的家庭网箱监测系统只能提供用于总睡眠时间的数据。此外, 尽管基于活动的家庭网箱监测的输出提供了有关睡眠时间的信息, 但由于四十年代间隔的固有限制, 我们无法准确评估时间长短, 因为3。尽管有这些限制, 家庭网箱监测睡眠持续时间提供了一个重要的生物措施, 可能会影响许多下游因素, 包括动物的健康和行为5。
基于活动的家庭网箱监测已被用来检测睡眠在许多研究表明其通用性。我们引用了这些研究的示例4,6,7,8,9,10,11,12。除了所介绍的方法外, 还有其他方法可以通过基于活动的监视来检测睡眠, 每个测试都包含自己的限制13,14。其中一些研究检查长时间的不间断睡眠 (72 小时), 而一些检查睡眠在块24小时。在这项研究中, 我们提出了每24小时的睡眠分析后, 对每日腹腔 (IP) 注射和周期性笼改变的小鼠模型的脆弱 X 综合征 (Fmr1高鼠)。我们选择了Fmr1高鼠, 因为它们减少了睡眠4 , 并且被假定为对感官信息15的超反应。我们的数据突出了在应对紧张事件时检测睡眠模式变化的能力。该方法适用于获取大鼠群中睡眠的一般信息。该方法有助于了解特定基因改变对睡眠的影响, 药理治疗的效果, 或对事件的反应, 如压力源。此外, 该方法提供了一个简单的方法来筛选的反应之前, 开始更多的参与研究。

<table><tbody><tr><td>Comprehensive Lab Animal Monitoring System (CLAMS)</td><td>Columbus Instruments</td><td></td><td>Equipment and software to analyze sleep duration</td></tr><tr><td>Captisol Research Grade</td><td>Captisol</td><td>RC-0C7-100</td><td>Captisol for dissolving hydrophobic compounds</td></tr><tr><td>30 G BD Needle 1/2 inch</td><td>BD</td><td>305106</td><td>Needle for injections</td></tr><tr><td>BD Disposable Syringes</td><td>Fisher</td><td>14-823-30</td><td>Syringes for injections</td></tr><tr><td>B6.129P2-Fmr1tm1Cgr/J</td><td>Jackson Labs</td><td>3025</td><td>Fmr1 KO mice</td></tr><tr><td>Super Mouse 750 Mouse Cage</td><td>Lab Products, Inc.&amp;nbsp;</td><td></td><td>Homecages for the mice</td></tr><tr><td>SANI-Chips Bedding</td><td>PJ Murphys</td><td></td><td>Bedding for the mice</td></tr></tbody></table>

noninvasive, high-throughput determination of sleep duration in rodents

传统上, 睡眠是由脑电图 (EEG) 监控的。啮齿类动物的脑电图研究需要手术后电极的植入, 然后是长时间的恢复期。为了执行脑电图记录, 动物被连接到一个接收器, 形成一个不自然的拴在头上安装。脑电图监测是耗时的, 对动物携带风险, 并不是一个完全自然的测量睡眠的设置。其他的方法来检测睡眠, 特别是在高通量的方式, 将极大地推动睡眠研究领域。在这里, 我们描述了一种通过基于活动的家庭网箱监测来检测睡眠的验证方法。以往的研究表明, 通过这种方法评估的睡眠与传统的基于脑电图的措施所定义的睡眠有很大程度的一致。虽然这种方法是对总睡眠时间的验证, 重要的是要注意, 睡眠周期应该由一个有更好的时间分辨率的脑电图评估。脑电图还可以区分快速眼球运动 (rem) 和非 REM 睡眠, 更详细地说明睡眠的确切性质。然而, 基于活动的睡眠确定可以用来分析多天未受干扰的睡眠, 并评估睡眠作为对急性事件 (如压力) 的反应。在这里, 我们展示了这个系统的力量, 以检测小鼠对日常腹腔注射的反应。

为了确定每日注射对睡眠的影响, 以及动物是否习惯注射, 我们每天在上午9:00 进行14天的 IP 注射 (光循环始于凌晨6:00), 并记录12个Fmr1 C57Bl/6J 小鼠的睡眠时间。我们使用了一个在主题的设计, 每一个动物注射正常生理盐水连续4天 (天 1-4), 然后30% 个环糊精连续十天 (天 5-14)。环糊精的选择, 因为它可以用来溶解疏水性化合物的药物管理, 我们有兴趣的是如何对车辆注射?...

Watch this Scientific Journal Video about Noninvasive, High-throughput Determination of Sleep Duration in Rodents at JoVE.com

Noninvasive, High-throughput Determination of Sleep Duration in Rodents

我们描述了一种高通量的测量睡眠的方法, 以活动为基础的家庭网箱监测。这种方法比传统的基于脑电图的方法具有优势。它在确定总睡眠持续时间方面得到了很好的验证, 可以成为监测人类疾病模型中睡眠的有力工具。

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 ...

noninvasive-high-throughput-determination-of-sleep-duration-in-rodents

Research

JoVE Journal

Behavior

7.8K 次观看.  National Institutes of Health. 我们描述了一种高通量的测量睡眠的方法, 以活动为基础的家庭网箱监测。这种方法比传统的基于脑电图的方法具有优势。它在确定总睡眠持续时间方面得到了很好的验证, 可以成为监测人类疾病模型中睡眠的有力工具。

视频: Noninvasive, High-throughput Determination of Sleep Duration in Rodents

Measuring Circadian and Acute Light Responses in Mice using Wheel Running Activity

Circadian rhythms are physiological functions that cycle over a period of approximately 24 hours (circadian- circa: approximate and diem: day)1, 2. They are responsible for timing our sleep/wake cycles and hormone secretion. Since this timing is not precisely 24-hours, it is synchronized to the solar day by light input. This is accomplished via photic input from the retina to the suprachiasmatic nucleus (SCN) which serves as the master pacemaker synchronizing peripheral clocks in other regions of the brain and peripheral tissues to the environmental light dark cycle3-7. The alignment of rhythms to this environmental light dark cycle organizes particular physiological events to the correct temporal niche, which is crucial for survival8. For example, mice sleep during the day and are active at night. This ability to consolidate activity to either the light or dark portion of the day is referred to as circadian photoentrainment and requires light input to the circadian clock9. Activity of mice at night is robust particularly in the presence of a running wheel. Measuring this behavior is a minimally invasive method that can be used to evaluate the functionality of the circadian system as well as light input to this system. Methods that will covered here are used to examine the circadian clock, light input to this system, as well as the direct influence of light on wheel running behavior.

This article will review methods that can be used to determine circadian function and light responsiveness in mice.

测量小鼠使用车轮运行活动的昼夜和急性对光反应

Circadian rhythms are physiological functions that cycle over a period of approximately 24 hours (circadian- circa: approximate and diem: day)1, 2.  They ...

科研

神经科学

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. ...

行为学

Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenetic Manipulation of Neuronal Activity in the Rodent Cerebral Cortex

Although the brain represents less than 5% of the body by mass, it utilizes approximately one quarter of the glucose used by the body at rest1. The function of non rapid eye movement sleep (NREMS), the largest portion of sleep by time, is uncertain. However, one salient feature of NREMS is a significant reduction in the rate of cerebral glucose utilization relative to wakefulness2-4. This and other findings have led to the widely held belief that sleep serves a function related to cerebral metabolism. Yet, the mechanisms underlying the reduction in cerebral glucose metabolism during NREMS remain to be elucidated.
	One phenomenon associated with NREMS that might impact cerebral metabolic rate is the occurrence of slow waves, oscillations at frequencies less than 4 Hz, in the electroencephalogram5,6. These slow waves detected at the level of the skull or cerebral cortical surface reflect the oscillations of underlying neurons between a depolarized/up state and a hyperpolarized/down state7. During the down state, cells do not undergo action potentials for intervals of up to several hundred milliseconds. Restoration of ionic concentration gradients subsequent to action potentials represents a significant metabolic load on the cell8; absence of action potentials during down states associated with NREMS may contribute to reduced metabolism relative to wake.
	Two technical challenges had to be addressed in order for this hypothetical relationship to be tested. First, it was necessary to measure cerebral glycolytic metabolism with a temporal resolution reflective of the dynamics of the cerebral EEG (that is, over seconds rather than minutes). To do so, we measured the concentration of lactate, the product of aerobic glycolysis, and therefore a readout of the rate of glucose metabolism in the brains of mice. Lactate was measured using a lactate oxidase based real time sensor embedded in the frontal cortex. The sensing mechanism consists of a platinum-iridium electrode surrounded by a layer of lactate oxidase molecules. Metabolism of lactate by lactate oxidase produces hydrogen peroxide, which produces a current in the platinum-iridium electrode. So a ramping up of cerebral glycolysis provides an increase in the concentration of substrate for lactate oxidase, which then is reflected in increased current at the sensing electrode. It was additionally necessary to measure these variables while manipulating the excitability of the cerebral cortex, in order to isolate this variable from other facets of NREMS.
	We devised an experimental system for simultaneous measurement of neuronal activity via the elecetroencephalogram, measurement of glycolytic flux via a lactate biosensor, and manipulation of cerebral cortical neuronal activity via optogenetic activation of pyramidal neurons. We have utilized this system to document the relationship between sleep-related electroencephalographic waveforms and the moment-to-moment dynamics of lactate concentration in the cerebral cortex. The protocol may be useful for any individual interested in studying, in freely behaving rodents, the relationship between neuronal activity measured at the electroencephalographic level and cellular energetics within the brain.

A procedure is described for manipulating the activity of cerebral cortical pyramidal neurons optogenetically while the electroencephalogram, electromyogram, and cerebral lactate concentration are monitored. Experimental recordings are performed on cable-tethered mice while they undergo spontaneous sleep/wake cycles. Optogenetic equipment is assembled in our laboratory; recording equipment is commercially available.

同时脑电图，实时测量的啮齿动物的大脑皮质中的神经元活动的乳酸浓度与Optogenetic的操作

Although the brain represents less than 5% of the body  by mass, it utilizes approximately one quarter of the glucose used by the body  at rest1. The ...

推进睡眠障碍治疗:一种基于果 蝇的筛查方法

High-Throughput Small Molecule Drug Screening For Age-Related Sleep Disorders Using Drosophila melanogaster

Sleep, an essential component of health and overall well-being, often presents challenges for older individuals who frequently experience sleep disorders characterized by shortened sleep duration and fragmented patterns. These sleep disruptions also correlate with an increased risk of various illnesses in the elderly, including diabetes, cardiovascular diseases, and psychological disorders. Unfortunately, existing drugs for sleep disorders are associated with significant side effects such as cognitive impairment and addiction. Consequently, the development of new, safer, and more effective sleep disorder medications is urgently needed. However, the high cost and lengthy experimental duration of current drug screening methods remain limiting factors.
This protocol describes a cost-effective and high-throughput screening method that utilizes Drosophila melanogaster, a species with a highly conserved sleep regulation mechanism compared to mammals, making it an ideal model for studying sleep disorders in the elderly. By administering various small compounds to aged flies, we can assess their effects on sleep disorders. The sleep behaviors of these flies are recorded using an infrared monitoring device and analyzed with the open-source data package Sleep and Circadian Analysis MATLAB Program 2020 (SCAMP2020). This protocol offers a low-cost, reproducible, and efficient screening approach for sleep regulation. Fruit flies, due to their short life cycle, low husbandry cost, and ease of handling, serve as excellent subjects for this method. As an illustration, Reserpine, one of the tested drugs, demonstrated the ability to promote sleep duration in elderly flies, highlighting the effectiveness of this protocol.

作者聚焦:使用 DAM 系统克服果蝇睡眠测量的挑战 

Presented is a protocol for high-throughput drug screening to improve sleep by monitoring the sleep behavior of fruit flies in an elderly Drosophila model.

使用黑腹果蝇对年龄相关睡眠障碍进行高通量小分子药物筛选

Sleep, an essential component of health and overall well-being, often presents challenges for older individuals who frequently experience sleep disorders ...

Polygraphic Recording Procedure for Measuring Sleep in Mice

Recording of the epidural electroencephalogram (EEG) and electromyogram (EMG) in small animals, like mice and rats, has been pivotal to study the homeodynamics and circuitry of sleep-wake regulation. In many laboratories, a cable-based sleep recording system is used to monitor the EEG and EMG in freely behaving mice in combination with computer software for automatic scoring of the vigilance states on the basis of power spectrum analysis of EEG data. A description of this system is detailed herein. Steel screws are implanted over the frontal cortical area and the parietal area of 1 hemisphere for monitoring EEG signals. In addition, EMG activity is monitored by the bilateral placement of wires in both neck muscles. Non-rapid eye movement (Non-REM; NREM) sleep is characterized by large, slow brain waves with delta activity below 4 Hz in the EEG, whereas a shift from low-frequency delta activity to a rapid low-voltage EEG in the theta range between 6 and 10 Hz can be observed at the transition from NREM to REM sleep. By contrast, wakefulness is identified by low- to moderate-voltage brain waves in the EEG trace and significant EMG activity.

The recording of electroencephalogram (EEG) and electromyogram (EMG) in freely behaving mice is a critical step to correlate behavior and physiology with sleep and wakefulness. The experimental protocol described herein provides a cable-based system for acquiring EEG and EMG recordings in mice.

印刷业记录程序在小鼠体内测量睡眠

Recording of the epidural electroencephalogram (EEG) and electromyogram (EMG) in small animals, like mice and rats, has been pivotal to study the ...

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 ...

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 ...

The Sleep Nullifying Apparatus: A Highly Efficient Method of Sleep Depriving Drosophila

Sleep homeostasis, the increase in sleep observed following sleep loss, is one of the defining criteria used to identify sleep throughout the animal kingdom. As a consequence, sleep deprivation and sleep restriction are powerful tools that are commonly used to provide insight into sleep function. Nonetheless, sleep deprivation experiments are inherently problematic in that the deprivation stimulus itself may be the cause of observed changes in physiology and behavior. Accordingly, successful sleep deprivation techniques should keep animals awake and, ideally, result in a robust sleep rebound without also inducing a large number of unintended consequences. Here, we describe a sleep deprivation technique for Drosophila melanogaster. The Sleep Nullifying Apparatus (SNAP) administers a stimulus every 10s to induce negative geotaxis. Although the stimulus is predictable, the SNAP effectively prevents &#62;95% of nighttime sleep even in flies with high sleep drive. Importantly, the subsequent homeostatic response is very similar to that achieved using hand-deprivation. The timing and spacing of the stimuli can be modified to minimize sleep loss and thus examine non-specific effects of the stimulus on physiology and behavior. The SNAP can also be used for sleep restriction and to assess arousal thresholds. The SNAP is a powerful sleep disruption technique that can be used to better understand sleep function.

The Sleep Nullifying Apparatus: A Highly Efficient Method of Sleep Depriving Drosophila

Sleep deprivation is a powerful tool to investigate sleep function and regulation. We describe a protocol to sleep deprive Drosophila using the Sleep Nullifying Apparatus, and to determine the extent of rebound sleep induced by deprivation.

睡眠无效装置:剥夺嗜睡者睡眠的高效方法

Sleep homeostasis, the increase in sleep observed following sleep loss, is one of the defining criteria used to identify sleep throughout the animal ...

无创高通量测定啮齿类动物睡眠时间

摘要

探索更多视频

测量小鼠使用车轮运行活动的昼夜和急性对光反应

小鼠幼犬慢性睡眠剥夺的温和处理方法

同时脑电图，实时测量的啮齿动物的大脑皮质中的神经元活动的乳酸浓度与Optogenetic的操作

使用黑腹果蝇对年龄相关睡眠障碍进行高通量小分子药物筛选

使用黑腹果蝇对年龄相关睡眠障碍进行高通量小分子药物筛选

使用黑腹果蝇对年龄相关睡眠障碍进行高通量小分子药物筛选

印刷业记录程序在小鼠体内测量睡眠

量化睡眠小鼠的光谱功率和心率的慢速动力学

监测小鼠睡眠/觉醒状态期间神经回路的光遗传学操作

睡眠无效装置:剥夺嗜睡者睡眠的高效方法