We thank Dr. Larry D. Frye for editorial help with this manuscript. This work was supported by Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research 24300129 (to M.L.), 25890005 (to Y.O.) and 26640025 (to Y.T.), the National Agriculture and Food Research Organization (to Y.U.), the World Premier International Research Center Initiative (WPI) from the Ministry of Education, Culture, Sports, Science, and Technology (to Y.O., Y.T., Y.U. and M.L.) and the Nestl&#233; Nutrition Council, Japan (to M.L.).

伦理声明:涉及动物主题程序已经批准在筑波大学的体制动物实验委员会。 1.准备电极和电缆脑电图/肌电图记录<ol><li>按照下列程序准备脑电/肌电图记录电极。 注意:电极是一次性的，并且只能用于1动物。认真规划所有连接器的接线配置。将标记的连接器正确的方向。 <ol><li>焊接一个4针接头的每个引脚到2厘米的不锈钢丝。简言之，把导线到销的一个端部，放置一个热烙铁到丝销接头，和熔体一些焊料以保证足够的刚焊接操作顺利进入关节。要小心，不要过多的热量适用于针;否则，绕销塑料就会熔化。 </li><li>焊锡的每个附连到销头的头部2导线的自由端的1.0毫米直径的不锈钢螺钉。简言之，把导线以下面的螺丝头的螺纹的自由端，放置一个热烙铁到线螺纹接头，和熔体一些焊料以保证足够的刚焊接操作顺利进入关节。在2线与螺丝作为脑电图记录电极，而2线无螺丝作为肌电记录电极。 </li><li>用剪刀剥离关闭1毫米绝缘的肌电电极年底增加了肌电图信号的质量。 </li><li>完全覆盖所有的焊接管脚用环氧树脂胶粘剂用细木棍或牙签，以减少在脑电图/肌电图记录的电噪声。 </li></ol></li><li>对于如下所述的滑环连接电极准备的电缆。该电缆可重复使用。 <ol><li>焊接一个4针FFC / FPC连接器的每个引脚有30厘米的扁平电缆的电线。简言之，把导线到销的剥离端，放置一个热烙铁上线脚关节和一些熔化焊锡，以确保刚够焊料运行顺利进入关节。 注意:选择扁平电缆，它是适合用于脑电图/肌电图记录实验动物笼的高度的长度。 </li><li>焊料卷曲套接字在扁平电缆的另一端的电线的前端。简言之，保持压接插座的导线的裸露自由端，放置一个热烙铁到丝窝接头和熔体一些焊料以保证足够的刚焊接操作顺利进入关节。 </li><li>将每个压接插座成4位压接壳体。 </li><li>完全覆盖用细木棍或牙签压接插座与环氧胶粘剂。 </li></ol></li></ol> 2.植入电极在小鼠头部（时间:约20分钟） <ol><li>在消毒手术前一个热点珠灭菌所有的外科手术工具。麻醉雄性小鼠（10 - 20周龄，20 - 30克）腹膜内注射戊巴比妥（50毫克/千克）。检查鼠标深受捏脚趾麻醉后，刮胡子的头部和颈部与快船队的头发。 </li><li>将鼠标移动到立体框架，并修复了2耳棒的头部。取适量涂抹于眼睛凡士林，以防止干燥。净化剃光皮肤用酒精和剪沿中线用手术刀，露出颅骨。剪辑肌肤保持手术区开放。 </li><li>通过使用硬质合金铣刀（钻头尺寸:0.8毫米直径），钻头2个孔进入颅骨，一个比额叶皮质区（1毫米前方前囟，1.5毫米横向于中线）和其他在顶区（ 1毫米先于拉姆达，1.5毫米横向中线）的右半球，根据Paxinos和富兰克林的立体坐标。23 </li><li>通过使用一个珠宝商的螺丝刀，在孔的地方不锈钢脑电图记录的螺钉，使2 - 2.5圈的每个SCRE瓦硬膜外定位在皮层。 注:请勿将螺丝钉过深，以防损伤脑组织。检查螺钉紧紧地固定在头骨。这是很重要的过程中的多个记录（通常，超过1个月）的长期具有稳定的EEG信号。威格利螺丝生产脑电图文物和可能的试验计划结束前脱落。 </li><li>固定电极组件（参见1.1节，销向上翻）与瞬间胶的头骨，并盖上牙科水泥。请与斜方肌（颈）的肌肉镊子双边小孔，插入充当肌电电极插入孔的不锈钢丝。缝合用丝线（0.1毫米直径）的皮肤，以避免肌肉的曝光。 </li><li>从立体框架取下鼠标。辖氨苄青霉素（100毫克/千克）和美洛昔康（1毫克/千克）经腹膜内给小鼠，以避免细菌感染并减少手术后疼痛，分别。柯EP鼠标上的隔热垫和监视它，直到它已经恢复足够的意识，以保持胸骨斜卧。房子单独恢复期间鼠标，以避免去除电极由其他动物，和管理美洛昔康（1毫克/千克）经腹膜内对手术后的第一天。 </li></ol> 3.记录和采集脑电/肌电数据<ol><li>后1周的恢复期，房子单独每只小鼠在实验笼放置在一个隔音录音室。保持23±1°C的环境温度和自动控制为12小时光照/ 12小时黑暗周期（开灯08:00，光照强度〜100勒克斯）。 </li><li>鼠标头部脑电图/肌电电极组件连接到记录电缆。确保记录电缆被连接到一个滑环（其目的是使鼠标的移动并不限于由电缆的扭转）和脑电图/ EMG信号放大器。过滤EEG / EMG信号（EEG，0.5-64赫兹;肌电图，16-64赫兹），数字化以128赫兹的采样速率由模拟-数字转换器（A / D）和最后一台计算机上运行的EEG /肌电图记录软件（表&#39;材料&#39;， 第 4项上记录从底部）。 </li><li> ，观察者鼠标2 - 录音室中3天。如果EEG / EMG记录包括腹腔内药品监督管理部门，轻轻地在给药时处理鼠标上的每一个习惯一天。 </li><li>随后，开始EEG / EMG记录软件（表&#39;材料&#39;，从下第 4项）。 <ol><li>选择"数据文件信息"选项卡，然后点击旁边的文件名框中。输入一个文件名，然后单击"保存"。 </li><li>选择"录制状态"选项卡，并选中所有脑电图/肌电图通道将被记录下来。 </li><li>在"录制条件"选项卡中选择采样频率（128赫兹）。 </li><li>检查选择的通道是个正常显示E&#39;频道信息"选项卡。 </li><li>选择"定时设置"选项卡，然后单击"监视器"，显示脑电图和肌电图。 </li><li>检查EEG / EMG信号显示正常。 </li><li>选择"定时设置"选项卡，设置时钟时间记录在"主定时器"区域的开头和结尾。 </li><li>点击"监视"中的"计时器设置"选项卡开始录制。 </li></ol></li><li>在基线记录 ​​的EEG / EMG信号（ 即 ，自由表现小鼠的睡眠/觉醒行为）和不同的处理条件（如，咖啡因管理或假治疗）在几天。安乐死小鼠腹膜内注射戊巴比妥（200毫克/千克）的最后试验天之后。 </li></ol>基于脑电/肌电图的数据行为状态4.计分<ol><li>启动软件脑电/肌电图分析（表&#39;材料&#39;，从底部4 个入口 ）。打开在步骤3中产生的脑电图/肌电图的原始数据（.kcd文件），点击"休眠"选项卡，选择大纪元时间。选择10秒。 </li><li>点击"休眠"选项卡，选择"多筛选"自动得分全部10秒的时期分为3个阶段（ 即 ，NREM和REM睡眠和觉醒），脑电图和肌电图和功率谱分析的幅度的基础上，的脑电图。3 </li><li>点击"F​​FT条件脑电图"。 </li><li>设置参数为功率谱分析[256基准点（相当于2秒的脑电图），汉宁窗函数，以及5光谱每个时期的平均] 3点击"确定"。 </li><li>点击"开始筛选&#39;开始自动筛选。打开得分数据（.RAF文件）。 </li><li>检查自动筛选结果，如果有必要，基于标准的标准手动纠正它们（见代表结果， 图1B 和表1 </s仲&gt;）。2，3简而言之，单击并按住不正确的拿下了划时代鼠标左键并拖动光标跨越的错误打进时代的字符串。松开鼠标左键，在弹出的窗口中选择正确的行为状态。 注意:有时，时代在两个警觉状态之间的转换也难以明确比分一种状态。在这种情况下，历元应评价的表面上的状态，并且在同一标准应该被应用到类似的历元在整个实验，以保证数据的再现性。 </li></ol>

<ol>
	<li>Berger, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=&#220;ber+das+Elektrenkephalogramm+des+Menschen.">&#220;ber das Elektrenkephalogramm des Menschen.</a> Arch. Psych. 87 (1), 527-570 (1929).</li><li>Tobler, I., Deboer, T., Fischer, M. <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+sleep+regulation+in+normal+and+prion+protein-deficient+mice.">Sleep and sleep regulation in normal and prion protein-deficient mice.</a> J. Neurosci. 17 (5), 1869-1879 (1997).</li><li>Kohtoh, 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=Algorithm+for+sleep+scoring+in+experimental+animals+based+on+fast+Fourier+transform+power+spectrum+analysis+of+the+electroencephalogram.">Algorithm for sleep scoring in experimental animals based on fast Fourier transform power spectrum analysis of the electroencephalogram.</a> Sleep Biol. Rhythm. 6 (3), 163-171 (2008).</li><li>Rosenbaum, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Warum m&#252;ssen wir schlafen? : eine neue Theorie des Schlafes. , (1892).</li><li>Kubota, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kuniomi+Ishimori+and+the+first+discovery+of+sleep-inducing+substances+in+the+brain.">Kuniomi Ishimori and the first discovery of sleep-inducing substances in the brain.</a> Neurosci. Res. 6 (6), 497-518 (1989).</li><li>Ishimori, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=True+cause+of+sleep:+a+hypnogenic+substance+as+evidenced+in+the+brain+of+sleep-deprived+animals.">True cause of sleep: a hypnogenic substance as evidenced in the brain of sleep-deprived animals.</a> Tokyo Igakkai Zasshi. 23, 429-457 (1909).</li><li>Legendre, R., Pieron, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recherches+sur+le+besoin+de+sommeil+cons&#233;cutif+&#224;+une+veille+prolong&#233;e.">Recherches sur le besoin de sommeil cons&#233;cutif &#224; une veille prolong&#233;e.</a> Z. Allegem. Physiol. 14, 235-262 (1913).</li><li>Inou&#233;, S., Honda, K., Komoda, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sleep+as+neuronal+detoxification+and+restitution.">Sleep as neuronal detoxification and restitution.</a> Behav. Brain. Res. 69 (1-2), 91-96 (1995).</li><li>Urade, Y., Hayaishi, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prostaglandin+D2+and+sleep/wake+regulation.">Prostaglandin D2 and sleep/wake regulation.</a> Sleep Med. Rev. 15 (6), 411-418 (2011).</li><li>Ueno, R., Ishikawa, Y., Nakayama, T., Hayaishi, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prostaglandin+D2+induces+sleep+when+microinjected+into+the+preoptic+area+of+conscious+rats.">Prostaglandin D2 induces sleep when microinjected into the preoptic area of conscious rats.</a> Biochem. Biophys. Res. Commun. 109 (2), 576-582 (1982).</li><li>Krueger, J. M., Walter, J., Dinarello, C. A., Wolff, S. M., Chedid, 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-promoting+effects+of+endogenous+pyrogen+(interleukin-1).">Sleep-promoting effects of endogenous pyrogen (interleukin-1).</a> Am. J. Physiol. 246 (6 Pt 2), R994-R999 (1984).</li><li>Porkka-Heiskanen, T., 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=Adenosine:+a+mediator+of+the+sleep-inducing+effects+of+prolonged+wakefulness.">Adenosine: a mediator of the sleep-inducing effects of prolonged wakefulness.</a> Science. 276 (5316), 1265-1268 (1997).</li><li>Garcia-Garcia, F., Acosta-Pena, E., Venebra-Munoz, A., Murillo-Rodriguez, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sleep-inducing+factors.">Sleep-inducing factors.</a> CNS Neurol. Disord. Drug. Targets. 8 (4), 235-244 (2009).</li><li>Huitron-Resendiz, 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=Urotensin+II+modulates+rapid+eye+movement+sleep+through+activation+of+brainstem+cholinergic+neurons.">Urotensin II modulates rapid eye movement sleep through activation of brainstem cholinergic neurons.</a> J. Neurosci. 25 (23), 5465-5474 (2005).</li><li>Wilkins, R. H., Brody, I. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Encephalitis+lethargica.">Encephalitis lethargica.</a> Arch. Neurol. 18 (3), 324-328 (1968).</li><li>von Economo, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Die+encephalitis+lethargica.">Die encephalitis lethargica.</a> Wien. Klin. Wochenschr. 30, 581-585 (1917).</li><li>Moruzzi, G., Magoun, H. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brain+stem+reticular+formation+and+activation+of+the+EEG.">Brain stem reticular formation and activation of the EEG.</a> Electroencephalogr. Clin. Neurophysiol. 1 (4), 455-473 (1949).</li><li>Saper, C. B., Fuller, P. M., Pedersen, N. P., Lu, J., Scammell, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sleep+state+switching.">Sleep state switching.</a> Neuron. 68 (6), 1023-1042 (2010).</li><li>Jones, B. E., Krnjevic , K., L, D. e. s. c. a. r. r. i. e. s., S, M. i. r. c. e. a. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Progress in Brain Research. 145, 157-169 (2004).</li><li>Saper, C. B., Scammell, T. E., Lu, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypothalamic+regulation+of+sleep+and+circadian+rhythms.">Hypothalamic regulation of sleep and circadian rhythms.</a> Nature. 437 (7063), 1257-1263 (2005).</li><li>Fort, P., Bassetti, C. L., Luppi, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alternating+vigilance+states:+new+insights+regarding+neuronal+networks+and+mechanisms.">Alternating vigilance states: new insights regarding neuronal networks and mechanisms.</a> Eur. J. Neurosci. 29 (9), 1741-1753 (2009).</li><li>Lu, J., Sherman, D., Devor, M., Saper, C. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+putative+flip-flop+switch+for+control+of+REM+sleep.">A putative flip-flop switch for control of REM sleep.</a> Nature. 441 (7093), 589-594 (2006).</li><li>Paxinos, G., Franklin, K. B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The mouse brain in stereotaxic coordinates. , (2001).</li><li>Lazarus, M., 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=Arousal+effect+of+caffeine+depends+on+adenosine+A2A+receptors+in+the+shell+of+the+nucleus+accumbens.">Arousal effect of caffeine depends on adenosine A2A receptors in the shell of the nucleus accumbens.</a> J. Neurosci. 31 (27), 10067-10075 (2011).</li><li>Huang, Z. L., 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=Adenosine+A2A,+but+not+A1,+receptors+mediate+the+arousal+effect+of+caffeine.">Adenosine A2A, but not A1, receptors mediate the arousal effect of caffeine.</a> Nat. Neurosci. 8 (7), 858-859 (2005).</li><li>Qu, W. M., Huang, Z. L., Xu, X. H., Matsumoto, N., Urade, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dopaminergic+D1+and+D2+receptors+are+essential+for+the+arousal+effect+of+modafinil.">Dopaminergic D1 and D2 receptors are essential for the arousal effect of modafinil.</a> J. Neurosci. 28 (34), 8462-8469 (2008).</li><li>Huang, Z. L., 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=Arousal+effect+of+orexin+A+depends+on+activation+of+the+histaminergic+system.">Arousal effect of orexin A depends on activation of the histaminergic system.</a> Proc. Natl. Acad. Sci. USA. 98 (17), 9965-9970 (2001).</li><li>Xu, Q., 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=A+mouse+model+mimicking+human+first+night+effect+for+the+evaluation+of+hypnotics.">A mouse model mimicking human first night effect for the evaluation of hypnotics.</a> Pharmacol. Biochem. Behav. 116, 129-136 (2014).</li><li>Cho, 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=Marine+polyphenol+phlorotannins+promote+non-rapid+eye+movement+sleep+in+mice+via+the+benzodiazepine+site+of+the+GABAA+receptor.">Marine polyphenol phlorotannins promote non-rapid eye movement sleep in mice via the benzodiazepine site of the GABAA receptor.</a> Psychopharmacol. 231 (14), 2825-2837 (2014).</li><li>Liu, Y. Y., 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=Piromelatine+exerts+antinociceptive+effect+via+melatonin,+opioid,+and+5HT1A+receptors+and+hypnotic+effect+via+melatonin+receptors+in+a+mouse+model+of+neuropathic+pain.">Piromelatine exerts antinociceptive effect via melatonin, opioid, and 5HT1A receptors and hypnotic effect via melatonin receptors in a mouse model of neuropathic pain.</a> Psychopharmacol. 231 (20), 3973-3985 (2014).</li><li>Qu, W. M., 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=Lipocalin-type+prostaglandin+D+synthase+produces+prostaglandin+D2+involved+in+regulation+of+physiological+sleep.">Lipocalin-type prostaglandin D synthase produces prostaglandin D2 involved in regulation of physiological sleep.</a> Proc. Natl. Acad. Sci. USA. 103 (47), 17949-17954 (2006).</li></ol>

The authors Yujiro Tauguchi and Sayaka Kohtoh are employees of Kissei Comtech Co., Ltd that develops SleepSign software for acquisition and automatic scoring of EEG/EMG data used in this article.

这个协议描述一个建立​​脑电/肌电图记录，允许睡眠和清醒的下低噪声，低成本，以及高通量条件的评估。由于脑电图/ EMG电极头组件的小尺寸，本系统可与其它植入物用于帧内脑实验，包括光遗传学（光纤植入），或结合同步插管植入，微注射的药物进入鼠标进行组合脑31。此外，电极头组件相对于多针头的设计提供了灵活性在记录信道数，如果需要的额外的电信号（例如，对...

技术的进步往往沉淀飞跃的神经生物学过程的理解。例如，汉斯Berger的发现于1929年，从人的头皮记录电势采取正弦波的形式，其频率直接与受试者的觉醒状态的水平，导致在睡眠觉醒的理解迅速进展法规，在动物和人类一样的。1为了这一天的electroencephlogram（EEG），与肌电图（EMG）， 即相结合，由骨骼肌产生的电活动，代表了几乎所有的实验和临床数据"脊梁"评估，即寻求相关行为和生理与皮质神经元的行为动物，包括人类的活动。在最基本的睡眠研究实验室这些脑电图记录是通过使用一个基于电缆的系统（图1），其中获得ð执行ATA进行离线到图案和频谱分析[例如，施加快速傅立叶变换（FFT）算法]来确定受试者的警觉状态被记录。2，第3睡眠由快速眼动（REM）和非快速眼动（NREM）睡眠。 REM睡眠的特征在于快速低压脑电图，随机眼球运动，和肌肉弛缓，一种状态，其中的肌肉得到有效瘫痪。 REM睡眠也被称为异相睡眠的，因为大脑活动类似于觉醒，而体是来自大脑的主要断开，似乎是在深度睡眠。与此相反，运动神经元期间NREM睡眠刺激但没有眼睛运动。人NREM睡眠可分为4个阶段，其中第4阶段被称为深睡眠或慢波睡眠，并与0.5之间的三角活动确定大的，缓慢的脑电波 - 4赫兹的脑电图。另一方面，在较小的动物NREM睡眠的相之间的细分，象大鼠次的小鼠，还没有建立，主要是因为它们在人类可见没有睡眠的长综合时间。 多年来，和脑电图解释的基础上，几种模式的睡眠觉醒调节，既电路交换和体液为基础，已经被提出。神经和睡眠的需要，或者细胞基础"睡眠驱动器，"仍然没有得到解决，但已被概念化为衡压力在清醒周期的建立，是由睡眠消散。有一种说法是，在觉醒，他们的点滴积累是睡眠衡压力托底内源性somnogenic因素的累积。虽然睡眠是通过体液因素的调控的第一个正式假说已记入出版罗森鲍姆的工作，于1892年4，这是石森5,6和 Pieron 7谁独立，并在100多年前，展现了促睡眠的化学物质的存在。这两个研究人员提出，事实上证明，这hypnogenic物质或"hypnotoxins"存在于睡眠剥夺狗的脑脊液（CSF）8近百年来牵连的睡眠自我平衡过程中的几个额外的假定hypnogenic物质已被确定（综述见文献9），包括前列腺素（PG）的D 2，10细胞因子，11腺苷，12花生四烯酸乙醇胺，13和尾加压素II肽。14 在初期和中期，20 世纪产生的结果，鼓舞睡眠和清醒和电路为基础的理论在一定程度上由Economo 15，16，Moruzzi和Magoun 17，和其他人的实验工作，盖过了当时的体液学说睡觉。到目前为止，几个"电路模型"已经提出，每一个通知不同的质量和数量的数据（综述见参考文献18）。一种模式例如提出，慢波睡眠是通过从胆碱能神经元在基底前脑，区域主要consisiting布罗卡的对角线频带和质inominata的水平肢的细胞核的腺苷介导的抑制乙酰胆碱的释放产生的。睡眠/觉醒第 19另一种流行的模型描述的触发器开关机构的睡眠诱发的神经元之间的腹外侧视前区和下丘脑和脑干相互抑制相互作用唤醒诱导的神经元基础上的。18，20，21此外，对于在进出REM睡眠的开关，一个类似的往复抑制相互作用已提出在脑干区域，即腹侧导水管周围灰质，横向脑桥被盖，和sublaterodorsal核。22总的来说，这些模型已经证明有价值启发式和提供重要的解释性框架，在睡眠的调查研究;然而，一个叶的分子机制和电路调节睡眠 - 觉醒周期的叔更全面的理解将需要其组件的更完整的认识。该系统为印刷业记录下文详述应该有助于这一目标。

<table border="0" cellpadding="0" cellspacing="0" width="729">
	<tbody>
		<tr height="19">
			<td height="19" style="height:19px;width:216px;">4-pin header</td>
			<td style="width:143px;">Hirose</td>
			<td style="width:167px;">A3B-4PA-2DSA(71)</td>
			<td style="width:205px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:216px;">Ampicillin</td>
			<td style="width:143px;">Meiji Seika</td>
			<td style="width:167px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Analog-to-digital converter</td>
			<td style="width:143px;">Contec</td>
			<td style="width:167px;">AD16-16U(PCIEV)</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Caffeine</td>
			<td style="width:143px;">Sigma</td>
			<td style="width:167px;">C0750</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Carbide cutter</td>
			<td style="width:143px;">Minitor</td>
			<td style="width:167px;">B1055</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:216px;">Crimp housing</td>
			<td style="width:143px;">Hirose</td>
			<td style="width:167px;">DF11-4DS-2C</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Crimp socket</td>
			<td style="width:143px;">Hirose</td>
			<td style="width:167px;">DF11-30SC</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Dental cement (Toughron Rebase)</td>
			<td style="width:143px;">Miki Chemical Product</td>
			<td style="width:167px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Epoxy adhesive</td>
			<td style="width:143px;">Konishi</td>
			<td style="width:167px;">#16351</td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:216px;">FFC/FPC connector</td>
			<td style="width:143px;">Honda Tsushin Kogyo</td>
			<td style="width:167px;">FFC-10BMEP1(B)</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Flat cable</td>
			<td style="width:143px;">Hitachi Cable</td>
			<td style="width:167px;">20528-ST LF</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Instant glue (Aron Alpha A)</td>
			<td style="width:143px;">Toagosei</td>
			<td style="width:167px;">N/A</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Meloxicam</td>
			<td style="width:143px;">Boehringer Ingelheim</td>
			<td style="width:167px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Pentobarbital</td>
			<td style="width:143px;">Kyoritsu Seiyaku</td>
			<td style="width:167px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Signal amplifier</td>
			<td style="width:143px;">Biotex</td>
			<td style="width:167px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Sleep recording chamber</td>
			<td style="width:143px;">APL</td>
			<td style="width:167px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">SleepSign software</td>
			<td style="width:143px;">Kissei Comtec</td>
			<td style="width:167px;">N/A</td>
			<td>for EEG/EMG recording/analysis</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Slip ring</td>
			<td style="width:143px;">Biotex</td>
			<td style="width:167px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Stainless steel screw</td>
			<td style="width:143px;">Yamazaki</td>
			<td style="width:167px;">N/A</td>
			<td>&#966;1.0&#215;2.0</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Stainless steel wire</td>
			<td style="width:143px;">Cooner Wire</td>
			<td style="width:167px;">AS633</td>
			<td></td>
		</tr>
	</tbody>
</table>

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.

图1B示出的鼠标脑电图在不同警觉状态的例子。如表 1所示，信号出现时间被分类为NREM睡眠如果脑电图显示缓慢的大脑电波低于4 Hz的δ节奏和肌电图仅具有弱或无信号。时期被列为REM睡眠，如果脑电图显示6和10赫兹之间的θ范围内迅速低电压脑电波和肌电图显示低振幅。其他历元应归类为觉醒（即，低到中等电压EEG和发生EMG活动的）。 <p...

Watch this Scientific Journal Video about Polygraphic Recording Procedure for Measuring Sleep in Mice at JoVE.com

Polygraphic Recording Procedure for Measuring Sleep in Mice

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 ...

polygraphic-recording-procedure-for-measuring-sleep-in-mice

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JoVE Journal

Anatomy and Physiology

Neuroscience

Unit 1

Functions of the Central and Peripheral Nervous System

SCIENTISTS IN ACTION

23.3K 次观看.  University of Tsukuba. 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.

视频: Polygraphic Recording Procedure for Measuring Sleep in Mice

Measuring Motor Coordination in Mice

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such as emotionality or cognition can be assessed, however, it is vital to determine whether the motor capabilities of e.g. a mutant or lesioned mouse allow such an assessment. Performance on a maze task requiring strength and coordination, such as the Morris water maze, might well be impaired in a mouse by motor, rather than cognitive, impairments, so it is essential to selectively dissect the latter from the former. For example, sensorimotor impairments caused by NMDA antagonists have been shown to impair water maze performance2. 
Motor coordination has traditionally been assessed in mice and rats by the rotarod test, in which the animal is placed on a horizontal rod that rotates about its long axis; the animal must walk forwards to remain upright and not fall off. Both set speed and accelerating versions of the rotarod are available.
The other three tests described in this article (horizontal bar, static rods and parallel bars) all measure coordination on static apparatus. The horizontal bar also requires strength for adequate performance, particularly of the forelimbs as the mouse initially grips the bar just with the front paws. 
Adult rats do not perform well on tests such as the static rods and parallel bars (personal observations); they appear less well coordinated than mice. I have only tested male rats, however, and male mice seem generally less well coordinated than females. Mice appear to have a higher strength:weight ratio than rats; the Latin name, Mus musculus, seems entirely appropriate. The rotarod, the variations of the foot fault test12 or the Catwalk (Noldus)15 apparatus are generally used to assess motor coordination in rats.

Protocols are presented for two established motor coordination tasks, the accelerating rotarod and horizontal bar, also two tests developed in Oxford recently, the static rods and parallel bars. These tests can detect motor impairments potentially of interest in their own right, as well as being possible variables in tests of other areas of behavior.

测量小鼠运动协调

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such ...

科研

神经科学

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from individual neurons are critical for understanding how neural circuits function under natural conditions. Traditionally, these recordings have been performed 'blind', meaning the identity of the recorded cell is unknown at the start of the recording. Cellular identity can be subsequently determined via intracellular1, juxtacellular2 or loose-patch3 iontophoresis of dye, but these recordings cannot be pre-targeted to specific neurons in regions with functionally heterogeneous cell types. Fluorescent proteins can be expressed in a cell-type specific manner permitting visually-guided single-cell electrophysiology4-6. However, there are many model systems for which these genetic tools are not available. Even in genetically accessible model systems, the desired promoter may be unknown or genetically homogenous neurons may have varying projection patterns. Similarly, viral vectors have been used to label specific subgroups of projection neurons7, but use of this method is limited by toxicity and lack of trans-synaptic specificity. Thus, additional techniques that offer specific pre-visualization to record from identified single neurons in vivo are needed. Pre-visualization of the target neuron is particularly useful for challenging recording conditions, for which classical single-cell recordings are often prohibitively difficult8-11. The novel technique described in this paper uses retrograde transport of a fluorescent dye applied using tungsten needles to rapidly and selectively label a specific subset of cells within a particular brain region based on their unique axonal projections, thereby providing a visual cue to obtain targeted electrophysiological recordings from identified neurons in an intact circuit within a vertebrate CNS.
The most significant novel advancement of our method is the use of fluorescent labeling to target specific cell types in a non-genetically accessible model system. Weakly electric fish are an excellent model system for studying neural circuits in awake, behaving animals12. We utilized this technique to study sensory processing by "small cells" in the anterior exterolateral nucleus (ELa) of weakly electric mormyrid fish. "Small cells" are hypothesized to be time comparator neurons important for detecting submillisecond differences in the arrival times of presynaptic spikes13. However, anatomical features such as dense myelin, engulfing synapses, and small cell bodies have made it extremely difficult to record from these cells using traditional methods11, 14. Here we demonstrate that our novel method selectively labels these cells in 28% of preparations, allowing for reliable, robust recordings and characterization of responses to electrosensory stimulation.

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde transport of fluorescent dye labels a sub-population of neurons based on anatomical projection. Labeled axons can be visually targeted in vivo, permitting extracellular recording from identified axons. This technique facilitates recording when neurons cannot be labeled through genetic manipulation or are difficult to isolate using 'blind' in vivo approaches.

逆行荧光染料标记允许有针对性的外单细胞记录技术鉴定神经元<em&gt;在体内</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, including rats, hamsters, and mice, are active during the night and relatively inactive during the day. Many other behavioral and physiological measures also exhibit daily rhythms, but in rodents, running-wheel activity serves as a particularly reliable and convenient measure of the output of the master circadian clock, the suprachiasmatic nucleus (SCN) of the hypothalamus. In general, through a process called entrainment, the daily pattern of running-wheel activity will naturally align with the environmental light-dark cycle (LD cycle; e.g. 12 hr-light:12 hr-dark). However circadian rhythms are endogenously generated patterns in behavior that exhibit a ~24 hr period, and persist in constant darkness. Thus, in the absence of an LD cycle, the recording and analysis of running-wheel activity can be used to determine the subjective time-of-day. Because these rhythms are directed by the circadian clock the subjective time-of-day is referred to as the circadian time (CT). In contrast, when an LD cycle is present, the time-of-day that is determined by the environmental LD cycle is called the zeitgeber time (ZT).
Although circadian rhythms in running-wheel activity are typically linked to the SCN clock6-8, circadian oscillators in many other regions of the brain and body9-14 could also be involved in the regulation of daily activity rhythms. For instance, daily rhythms in food-anticipatory activity do not require the SCN15,16 and instead, are correlated with changes in the activity of extra-SCN oscillators17-20. Thus, running-wheel activity recordings can provide important behavioral information not only about the output of the master SCN clock, but also on the activity of extra-SCN oscillators. Below we describe the equipment and methods used to record, analyze and display circadian locomotor activity rhythms in laboratory rodents.

Circadian rhythms in voluntary wheel-running activity in mammals are tightly coupled to the molecular oscillations of a master clock in the brain. As such, these daily rhythms in behavior can be used to study the influence of genetic, pharmacological, and environmental factors on the functioning of this circadian clock.

运行轮活动在啮齿类动物的昼夜节律的记录和分析

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, ...

Measuring Spinal Presynaptic Inhibition in Mice By Dorsal Root Potential Recording In Vivo

Presynaptic inhibition is one of the most powerful inhibitory mechanisms in the spinal cord. The underlying physiological mechanism is a depolarization of primary afferent fibers mediated by GABAergic axo-axonal synapses (primary afferent depolarization). The strength of primary afferent depolarization can be measured by recording of volume-conducted potentials at the dorsal root (dorsal root potentials, DRP). Pathological changes of presynaptic inhibition are crucial in the abnormal central processing of certain pain conditions and in some disorders of motor hyperexcitability. Here, we describe a method of recording DRP in vivo in mice. The preparation of spinal cord dorsal roots in the anesthetized animal and the recording procedure using suction electrodes are explained. This method allows measuring GABAergic DRP and thereby estimating spinal presynaptic inhibition in the living mouse. In combination with transgenic mouse models, DRP recording may serve as a powerful tool to investigate disease-associated spinal pathophysiology. In vivo recording has several advantages compared to ex vivo isolated spinal cord preparations, e.g. the possibility of simultaneous recording or manipulation of supraspinal networks and induction of DRP by stimulation of peripheral nerves.

Measuring Spinal Presynaptic Inhibition in Mice By Dorsal Root Potential Recording In Vivo

GABAergic presynaptic inhibition is a powerful inhibitory mechanism in the spinal cord important for motor and sensory signal integration in spinal cord networks. Underlying primary afferent depolarization can be measured by recording of dorsal root potentials (DRP). Here we demonstrate a method of in vivo recording of DRP in mice.

测量脊髓突触前抑制小鼠背根电位记录<em&gt;在体内</em

Presynaptic inhibition is one of the most powerful inhibitory mechanisms in the spinal cord. The underlying physiological mechanism is a depolarization of ...

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life. The cold plantar assay allows the objective and inexpensive assessment of cold sensitivity in mice, and can quantify both analgesia and hypersensitivity. Mice are acclimated on a glass plate, and a compressed dry ice pellet is held against the glass surface underneath the hindpaw. The latency to withdrawal from the cooling glass is used as a measure of cold sensitivity.
Cold sensation is also important for survival in regions with seasonal temperature shifts, and in order to maintain sensitivity animals must be able to adjust their thermal response thresholds to match the ambient temperature. The Cold Plantar Assay (CPA) also allows the study of adaptation to changes in ambient temperature by testing the cold sensitivity of mice at temperatures ranging from 30 &#176;C to 5 &#176;C. Mice are acclimated as described above, but the glass plate is cooled to the desired starting temperature using aluminum boxes (or aluminum foil packets) filled with hot water, wet ice, or dry ice. The temperature of the plate is measured at the center using a filament T-type thermocouple probe. Once the plate has reached the desired starting temperature, the animals are tested as described above.
This assay allows testing of mice at temperatures ranging from innocuous to noxious. The CPA yields unambiguous and consistent behavioral responses in uninjured mice and can be used to quantify both hypersensitivity and analgesia. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

The Cold Plantar Assay (CPA) measures cold responsiveness between 30 &#176;C and 5 &#176;C, and can also measure cold adaptation. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

测定冷灵敏度和适应小鼠一种简单廉价的方法

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

Extracellular Recording of Neuronal Activity Combined with Microiontophoretic Application of Neuroactive Substances in Awake Mice

Differences in the activity of neurotransmitters and neuromodulators, and consequently different neural responses, can be found between anesthetized and awake animals. Therefore, methods allowing the manipulation of synaptic systems in awake animals are required in order to determine the contribution of synaptic inputs to neuronal processing unaffected by anesthetics. Here, we present methodology for the construction of electrodes to simultaneously record extracellular neural activity and release multiple neuroactive substances at the vicinity of the recording sites in awake mice. By combining these procedures, we performed microiontophoretic injections of gabazine to selectively block GABAA receptors in neurons of the inferior colliculus of head-restrained mice. Gabazine successfully modified neural response properties such as the frequency response area and stimulus-specific adaptation. Thus, we demonstrate that our methods are suitable for recording single-unit activity and for dissecting the role of specific neurotransmitter receptors in auditory processing.
The main limitation of the described procedure is the relatively short recording time (~3 hr), which is determined by the level of habituation of the animal to the recording sessions. On the other hand, multiple recording sessions can be performed in the same animal. The advantage of this technique over other experimental procedures used to manipulate the level of neurotransmission or neuromodulation (such as systemic injections or the use of optogenetic models), is that the drug effect is confined to the local synaptic inputs to the target neuron. In addition, the custom-manufacture of electrodes allows adjustment of specific parameters according to the neural structure and type of neuron of interest (such as the tip resistance for improving the signal-to-noise ratio of the recordings).

We present methods for the construction of electrodes to simultaneously record extracellular neural activity and release multiple neuroactive substances at the vicinity of the recording sites in awake mice. This technique allows the detailed analysis of putative local synaptic inputs to the neuron of interest.

神经元电活动外记录与神经活性物质在清醒小鼠微电泳应用相结合

Differences in the activity of neurotransmitters and neuromodulators, and consequently different neural responses, can be found between anesthetized and ...

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons

The Xenopus tadpole retinotectal circuit, comprised of the retinal ganglion cells (RGCs) in the eye which form synapses directly onto neurons in the optic tectum, is a popular model to study how neural circuits self-assemble. The ability to carry out whole cell patch clamp recordings from tectal neurons and to record RGC-evoked responses, either in vivo or using a whole brain preparation, has generated a large body of high-resolution data about the mechanisms underlying normal, and abnormal, circuit formation and function. Here we describe how to perform the in vivo preparation, the original whole brain preparation, and a more recently developed horizontal brain slice preparation for obtaining whole cell patch clamp recordings from tectal neurons. Each preparation has unique experimental advantages. The in vivo preparation enables the recording of the direct response of tectal neurons to visual stimuli projected onto the eye. The whole brain preparation allows for the RGC axons to be activated in a highly controlled manner, and the horizontal brain slice preparation allows recording from across all layers of the tectum.

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons

In this paper, we discuss three brain preparations used for whole cell patch clamp recording to study the retinotectal circuit of Xenopus laevis tadpoles. Each preparation, with its own specific advantages, contributes to the experimental tractability of the Xenopus tadpole as a model to study neural circuit function.

爪蟾Tectal 神经元全细胞膜片钳记录的制备与协议

The Xenopus tadpole retinotectal circuit, comprised of the retinal ganglion cells (RGCs) in the eye which form synapses directly onto neurons in the optic ...

Recording Spatially Restricted Oscillations in the Hippocampus of Behaving Mice

The local field potential (LFP) emerges from ion movements across neural membranes. Since the voltage recorded by LFP electrodes reflects the summed electrical field of a large volume of brain tissue, extracting information about local activity is challenging. Studying neuronal microcircuits, however, requires a reliable distinction between truly local events and volume-conducted signals originating in distant brain areas. Current source density (CSD) analysis offers a solution for this problem by providing information about current sinks and sources in the vicinity of the electrodes. In brain areas with laminar cytoarchitecture such as the hippocampus, one-dimensional CSD can be obtained by estimating the second spatial derivative of the LFP. Here, we describe a method to record multilaminar LFPs using linear silicon probes implanted into the dorsal hippocampus. CSD traces are computed along individual shanks of the probe. This protocol thus describes a procedure to resolve spatially restricted neuronal network oscillations in the hippocampus of freely moving mice.

This protocol describes the recording of local field potentials with multi-shank linear silicon probes. Conversion of the signals using current source density analysis allows the reconstruction of local electrical activity in the mouse hippocampus. With this technique, spatially restricted brain oscillations can be studied in freely moving mice.

行为小鼠海马空间受限振荡的记录

The local field potential (LFP) emerges from ion movements across neural membranes. Since the voltage recorded by LFP electrodes reflects the summed ...

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 ...

Evaluation of Hemisphere Lateralization with Bilateral Local Field Potential Recording in Secondary Motor Cortex of Mice

This article demonstrates complete, detailed procedures for both in vivo bilateral recording and analysis of local field potential (LFP) in the cortical areas of mice, which are useful for evaluating possible laterality deficits, as well as for assessing brain connectivity and coupling of neural network activities in rodents. The pathological mechanisms underlying Alzheimer&#39;s disease (AD), a common neurodegenerative disease, remain largely unknown. Altered brain laterality has been demonstrated in aging people, but whether or not abnormal lateralization is one of the early signs of AD has not been determined. To investigate this, we recorded bilateral LFPs in 3-5-month-old AD model mice, APP/PS1, together with littermate wild type (WT) controls. The LFPs of the left and right secondary motor cortex (M2), specifically in the gamma band, were more synchronized in APP/PS1 mice than in WT controls, suggesting a declined hemispheric asymmetry of bilateral M2 in this AD mouse model. Notably, the recording and data analysis processes are flexible and easy to carry out, and can also be applied to other brain pathways when conducting experiments that focus on neuronal circuits.

We present in vivo electrophysiological recording of the local field potential (LFP) in bilateral secondary motor cortex (M2) of mice, which can be applied to evaluate hemisphere lateralization. The study revealed altered levels of synchronization between the left and right M2 in APP/PS1 mice compared to WT controls.

小鼠二次运动皮质中双边局部场电位记录半球横向化评价

This article demonstrates complete, detailed procedures for both in vivo bilateral recording and analysis of local field potential (LFP) in the cortical ...

印刷业记录程序在小鼠体内测量睡眠

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