Name: chronic sleep deprivation in mouse pups by means of gentle handling
Uploaded: 2018-10-11T13:30:00
Description: Watch this Scientific Journal Video about Chronic Sleep Deprivation in Mouse Pups by Means of Gentle Handling at JoVE.com

这项研究是由镍氢 (MH00889) 的内科学研究计划资助的。RMS 也得到了 FRAXA 博士后奖学金的支持。

注: 所有程序均经国家精神卫生动物护理和使用委员会批准, 并按照国家卫生研究院关于动物照料和使用的指导方针进行。
1. 睡眠剥夺设置
<ol><li>使用软画笔在睡觉时轻轻地戳老鼠。<ol>
<li>确保画笔可以通过鼠标笼进纸器的条形进行安装, 以确保小鼠在睡眠剥夺期间能够获得食物和水广告随意。</li>
		<li>不要将画笔用于多个保持架, 以防止在保持架之间传递气味。我们?...

<ol>
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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sleep+and+pain:+A+relationship+that+begins+in+early+life.">Sleep and pain: A relationship that begins in early life.</a> Pain Physician. 17 (6), E787-E798 (2014).</li><li>Hairston, I. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sleep+deprivation+effects+on+growth+factor+expression+in+neonatal+rats:+a+potential+role+for+BDNF+in+the+mediation+of+delta+power.">Sleep deprivation effects on growth factor expression in neonatal rats: a potential role for BDNF in the mediation of delta power.</a> J Neurophysiol. 91 (4), 1586-1595 (2004).</li><li>Hairston, I. 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在这里, 我们描述了一种睡眠剥夺, 温和的处理方法, 可用于啮齿动物幼崽和成人。轻柔的处理要求研究人员在睡眠剥夺期期间观察老鼠, 并在不活动或抽搐时轻轻地刺激动物以防止睡眠。以前的睡眠剥夺研究通过同步脑电图 (EEG) 记录进行验证, 轻柔处理可防止 rem 和非 rem (NREM) 睡眠10。虽然温和的处理是唯一可以用于幼鼠幼崽的睡眠剥夺方法之一, 但它的使用有一些局限性。
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睡眠在大脑发育过程中对神经元可塑性和突触形成和消除起着至关重要的作用1,2。具体地说, 快速眼动 (REM) 睡眠对于通过加强特定突触和突触修剪2形成稳定的突触电路至关重要。在生命早期的药理睡眠剥夺导致许多生理和行为缺陷3,4。除了药理性睡眠剥夺, 其他形式的睡眠剥夺, 如恒定摇晃, 强迫运动, 或厌恶刺激的表现, 在成年时与抑郁症状的症状相关, 不同的神经激活模式, 以及睡眠时间和睡眠连续性的变化在剥夺5,6,7后的恢复期间。当睡眠在小鼠的长期限制在开发期间超过五周, 行为缺陷被观察后一个月的睡眠恢复8。这些研究表明, 新生啮齿动物的睡眠中断会影响以后的睡眠模式、大脑功能和行为。强调睡眠对人类正常大脑发育的重要性, 许多神经发育障碍患者, 包括自闭症谱系障碍, 结节性硬化症, 易碎 X 综合征, 和注意缺陷/多动紊乱, 报告在睡眠模式的反常性除了各种行为缺陷1。
考虑到与异常睡眠模式相关的神经发育紊乱的数量, 了解睡眠不足对大脑功能和行为的影响是很重要的。然而, 尽管数据支持睡眠在发展过程中的重要性, 但大多数限制或剥夺睡眠动物的方法都集中在成人身上。这些技术包括各种测试, 需要强迫运动活动 (例如,恒定跑步机), 持续中断 (例如,旋转清扫棒或持续新颖的对象演示), 或扰乱动物的起始REM 睡眠 (例如,平台在水上)9,10。虽然这些方法有效地显示了与成人睡眠剥夺相关的问题, 但这些方法都不适合啮齿类动物, 因为它们的流动性有限, 睡眠率高, 以及生活中早期压力的负面影响。由于睡眠在发育过程中的重要性, 了解新生儿幼崽的睡眠限制对成年期的未来行为有何影响至关重要。
一种限制可用于啮齿动物幼崽的睡眠的技术是温和处理。温和的处理涉及调查人员与啮齿动物直接互动的任何时间睡眠行为观察。调查人员可以使用他们的手, 画笔, 或演示新的对象来物理扰乱休眠在新生儿啮齿动物8,10,11,12,13,14.睡眠可以行为由缺乏运动活动, 肌阵挛抽搐和闭眼 (在老啮齿动物幼崽) 确定。脑电图验证证实, 根据这些行为, 以温和的处理中断睡眠, P12 大鼠14的总睡眠时间减少91%。其他用于剥夺幼鼠的技术包括电击和不愉快刺激的表现, 但这些技术不能用于慢性睡眠限制, 比温和处理7更有压力,15. 慢性睡眠限制也可以通过振动协议来完成, 可以与脑电图电极植入和相关的计算机软件一起使用, 但此协议不会阻止所有睡眠, 因此更少控制比温和处理4,16。在没有特定设备、计算机软件或电极植入的情况下连续数天限制或剥夺小鼠睡眠的能力是温和处理技术的一个好处。通过轻柔处理的慢性睡眠限制有效地限制了新生啮齿动物的睡眠, 并在睡眠剥夺期8、11、12之后产生各种行为变化,17。
在这里, 我们描述的结果, 其中慢性睡眠剥夺三小时每天从产后 5 (P5) 到 P42 显著影响小鼠的社交性, 在30天的恢复期内, 睡眠不受干扰。这些结果突显了长期睡眠限制在发展过程中对未来行为的长远影响。

<table>
	<tbody>
		<tr>
			<td>C57BL/6J mice</td>
			<td>Jackson Labs</td>
			<td>000664</td>
			<td>Wildtype mice used</td>
		</tr>
		<tr>
			<td>Super Mouse 750 Mouse Cage</td>
			<td>Lab Products, Inc.&#160;</td>
			<td></td>
			<td>Cages for the mice</td>
		</tr>
		<tr>
			<td>SANI-Chips Bedding</td>
			<td>PJ Murphys</td>
			<td></td>
			<td>Bedding for the mice</td>
		</tr>
		<tr>
			<td>S&#38;S Bristle Brush Assortment Pack</td>
			<td>Staples</td>
			<td>Item # 13764, Model AB37100</td>
			<td>Paint brushes are used to gently poke mice</td>
		</tr>
	</tbody>
</table>

chronic sleep deprivation in mouse pups by means of gentle handling

睡眠对于正确的发育和神经可塑性至关重要。此外, 异常睡眠模式是许多神经发育紊乱的特征。研究发展过程中慢性睡眠限制对成人行为的影响可能会增加我们对神经发育紊乱行为症状出现的理解。虽然有许多方法可以用来限制在啮齿类动物的睡眠, 包括强迫运动, 持续中断, 厌恶刺激的表现, 或电击, 许多这些方法是非常紧张的, 不能用于新生小鼠。在这里, 我们描述温和的处理, 睡眠剥夺技术, 可以长期使用在整个发展和成年, 以达到睡眠限制。轻柔的处理包括在睡眠剥夺期间对小鼠进行密切观察, 并要求研究者在不活跃或显示与睡眠相关的行为时, 轻轻地向动物进行刺激。加上脑电图记录, 轻柔的处理可用于选择性地扰乱睡眠的特定阶段, 如快速眼动 (REM) 睡眠。温和的处理技术是一个强大的工具, 研究慢性睡眠限制的影响, 甚至在新生小鼠, 绕过许多更紧张的程序, 用于睡眠剥夺。

为了研究慢性睡眠限制对行为的影响, 我们每天从 P5 到 P428的上午11:00 和下午2:00 之间睡眠限制小鼠三小时。在睡眠限制后, 老鼠被不受干扰地留了四周, 并允许从睡眠限制中恢复。在四周的恢复期后, 对小鼠进行了行为测试。三室社会行为测试用于评估社交 (与新的物体相比, 对新老鼠的偏爱) 和社会新奇 (与熟悉的老鼠相比, 对新老鼠的偏爱)。在社交测试?...

Watch this Scientific Journal Video about Chronic Sleep Deprivation in Mouse Pups by Means of Gentle Handling at JoVE.com

Chronic Sleep Deprivation in Mouse Pups by Means of Gentle Handling

我们描述一种被称为温和处理的睡眠剥夺技术, 调查人员在任何时候都轻轻地对老鼠进行睡眠行为观察。这种方法是一个强大的工具, 让研究人员研究慢性睡眠限制在整个发展可能对未来的大脑生理学和行为的影响。

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. ...

这种方法可以帮助回答睡眠领域的关键问题，如产后发育过程中慢性睡眠限制的影响。这种技术的主要优点是，它的压力比它的其他睡眠剥夺技术小，可以很容易地在整个发展过程中使用，并持续到成年。首先将每个笼子分配给睡眠剥夺或对照组。确保小鼠能够获得食物和水，并移除任何可能限制小鼠视野的食物、床上用品或富集物。对于对照组，在产后第五天从育种者笼中取下陛下，然后与幼崽一起离开大坝，直到打奶。接下来，将分配给对照组的小鼠移动到睡眠剥夺室，并在实验期间每天用画笔连续用画笔轻轻推动小鼠10分钟。10分钟后，将老鼠返回动物室，不要打扰它们。首先在产后第五天将陛下从饲养员笼中移走，然后与幼崽一起离开大坝，直到打奶。在光周期内，每天将分配给睡眠剥夺组的小鼠移动到睡眠剥夺室，在那里进行监测。在此期间，如果怀疑鼠标睡着了，则用画笔轻轻抚摸鼠标，直到观察到反应。或者，反转小鼠，推上它们的背部，扰乱睡眠。如果出现以下任何情况，则考虑让鼠标处于睡眠状态：抽搐、不活动或眼睛闭着。如果老鼠四处走动，试图在背上翻过来，或者打扮自己，那么它们就清醒了。如果大坝在睡眠不足期间覆盖幼崽，则轻轻地将她从幼崽中推离，这样幼崽的视野是不受限制的。继续使用睡眠剥夺协议，直到实验终结点。当三个小时的睡眠剥夺期完成时，将床上用品和任何可能被取出的食物放回笼子，然后盖回笼子上。最后，将笼子回到动物的持有室，直到第二天睡眠不足。经过一个月的恢复期后，结果显示，在社交能力测试中，睡眠受限女性比控制雌性多时间嗅探陌生人的老鼠，睡眠受限的男性与对照男性相比，有花更多的时间嗅探物体。此外，在社会新奇测试阶段，睡眠受限的雌性小鼠比雌性对等对等小鼠花费更多的时间嗅探新小鼠。在尝试此程序时，重要的是要记住注意并及时响应睡眠前的行为，尤其是在睡眠需求很高的新生儿小鼠中。按照这个程序，其他方法，如开放领域，旋转，高架加迷宫，和社会行为测试可以执行，以回答其他问题，如慢性睡眠限制在发育过程中的长期行为影响。

Research

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

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 ...

科研

行为学

The Use of the Puzzle Box as a Means of Assessing the Efficacy of Environmental Enrichment

该拼图方块的使用作为评价环境富集的功效的一种手段

Environmental enrichment can dramatically influence the development and function of neural circuits.  Further, enrichment has been shown to successfully ...

Errors as a Means of Reducing Impulsive Food Choice

作为错误脉冲减少食物选择的一种手段

Nowadays, the increasing incidence of eating disorders due to poor self-control has given rise to increased obesity and other chronic weight problems, and ...

Manipulation of Epileptiform Electrocorticograms (ECoGs) and Sleep in Rats and Mice by Acupuncture

Manipulation of Epileptiform Electrocorticograms ECoGs and Sleep in Rats and Mice by Acupuncture

癫痫Electrocorticograms（ECoGs）的操作，并睡在针刺大鼠和小鼠

Ancient Chinese literature has documented that acupuncture possesses efficient therapeutic effects on epilepsy and insomnia. There is, however, little ...

Neurodevelopmental Reflex Testing in Neonatal Rat Pups

神经发育复检新生幼鼠

Neurodevelopmental reflex testing is commonly used in clinical practice to assess the maturation of the nervous system. Neurodevelopmental reflexes are ...

Noninvasive, High-throughput Determination of Sleep Duration in Rodents

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 ...

Measuring Active and Passive Tameness Separately in Mice

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 ...

Automated Measurements of Sleep and Locomotor Activity in Mexican Cavefish

墨西哥河鱼睡眠和睡眠活动的自动测量

Across phyla, sleep is characterized by highly conserved behavioral characteristics that include elevated arousal threshold, rebound following sleep ...

Collecting Sleep, Circadian, Fatigue, and Performance Data in Complex Operational Environments

在复杂操作环境中收集睡眠、循环、疲劳和性能数据

Sleep loss and circadian misalignment contribute to a meaningful proportion of operational accidents and incidents. Countermeasures and work scheduling ...

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

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 ...