这项工作得到了中国自然科学基金（31171355）和广东省自然科学基金资助（S2011010003403，2014A030313440）的支持。

动物护理，外科手术和记录程序符合华南师范大学动物护理与使用委员会的指引。 电极制备（图1A-C） 注意:计算机引脚位置只是通信设备中作为信号接口的一部分的插脚式接点。它由插入母连接器的公连接器组成。 <ol><li>借助于钳子，将公针和母针与计算机引脚位置（ 图1A ）分开。将公母针连接在一起形成电极并施加氰基丙烯酸酯以产生强的粘结（ 图1C ）。 </li><li>将电极放入装有蒸馏水的烧杯中，并将其放入超声波清洗机中10分钟。将它们移动到45℃的烘箱中30分钟。使用UV光灭菌电极30分钟。 </li></ol>外科手术（图1D-F） <ol><li>准备消毒的外科器械和立体定位仪。使用异氟醚麻醉（2.5％）与空气麻醉新生大鼠。当幼仔深度麻醉时，将异氟烷的剂量调整至1.0％。在手术前进行尾部或脚趾捏，以确保适当的麻醉深度。 </li><li>通过将耳棒放入耳道并稍微拧紧，将小狗的头部固定在立体定位装置中。 注意:不要过度收紧耳塞，因为新生儿头骨很软。 </li><li>通过喷洒70％乙醇的所有设备来维持无菌手术领域。使用手术刀在头部做一个15毫米的切口。使用镊子，轻轻地将头皮远离四角的中线。将一些盐水浸泡在皮肤下方，以保持切口大开（ 图1D ）。 </li><li>找到头骨上的阳光和喇叭点并用铅笔标记。使用注射器针头（26 G）在前额叶皮质（PFC）和海马中制造两个毛刺孔。 注意:PFC位于前囟后+1.8毫米，中线为-0.5毫米，而海马位于前囟前方-2.0毫米，中线横向为±0.5毫米（ 图1D和E ）。电极的深度不应大于皮质表面以下2毫米，以减少脑损伤。 </li><li>使用镊子保持电极并将参考和记录电极分别插入PFC和海马。在电极周围涂抹红霉素软膏，以避免任何可能的感染。使用氰基丙烯酸酯固定电极。 </li><li>准备牙科丙烯酸水泥，使其具有胶粘和粘稠的稠度。应用牙科水泥覆盖电极和头骨的其余部分。 注意:在应用牙科水泥之前彻底干燥颅骨。 </li&gt; <li>在电极上涂抹5％苦味酸以保护它们。 注意:整个程序应在生物安全罩中进行，以保持无菌条件。 </li><li>从立体定位框架中取出动物，并皮下注射300μL的10％葡萄糖。将其放在加热的毯子上以进行恢复。确保动物温暖（37°C）和动态（ 即完全恢复）。腹膜内给予丁丙诺啡（0.05 mg / kg）用于手术后疼痛。 注意:在重新获得足够的意识（即正常行为和运动）之前，不要将动物无人值守。 </li><li>在恢复意识之后，将小狗带回他们的水坝。等待两天，直到动物完全康复。 </li></ol> 3.脑电录音<ol><li>完全恢复后，将植入颅骨上的电极连接到放大器的笼中。将放大器连接到模拟数字c转换器并将转换器连接到计算机;应仔细对待连接线，以免缠结。 </li><li>在数据采集单元上选择至少10,000 Hz采样率进行记录（发送器的带宽为1-100 Hz）。确保数据正确采样。 </li><li>在进行基线记录后，用红藻氨酸（KA）（2mg / kg）腹膜内注射小鼠以诱导癫痫发作。注射后15分钟观察并记录癫痫放电。通过KA产生的缉获物通常是物理的。 注意:发炎持续时间约为15.2±0.9秒，发作持续时间约为62±5秒。通过IP注射水合氯醛（400 mg / kg）可以预防新生大鼠的癫痫发作。 </li><li>保存数字化数据并使用信号处理软件包（如spike2）进行分析。通过perf显示新生儿EEG信号中不同频率成分的功率水平功率谱分析。通过找到1到100 Hz（EEG频带） 9的均方根幅度，计算1分钟时间内的功率。 </li></ol>

<ol>
	<li>Pereda, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+synapses+and+their+functional+interactions+with+chemical+synapses.">Electrical synapses and their functional interactions with chemical synapses.</a> Nat Rev Neurosci. 15 (4), 250-263 (2014).</li><li>Chorev, E., Epsztein, J., Houweling, A. R., Lee, A. K., Brecht, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+recordings+from+behaving+animals--going+beyond+spikes.">Electrophysiological recordings from behaving animals--going beyond spikes.</a> Curr Opin Neurobiol. 19 (5), 513-519 (2009).</li><li>Freeborn, D. L., McDaniel, K. L., Moser, V. C., Herr, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+electroencephalography+(EEG)+to+assess+CNS+changes+produced+by+pesticides+with+different+modes+of+action:+effects+of+permethrin,+deltamethrin,+fipronil,+imidacloprid,+carbaryl,+and+triadimefon.">Use of electroencephalography (EEG) to assess CNS changes produced by pesticides with different modes of action: effects of permethrin, deltamethrin, fipronil, imidacloprid, carbaryl, and triadimefon.</a> Toxicol Appl Pharmacol. 282 (2), 184-194 (2015).</li><li>Werhahn, K. J., Hartl, E., Hamann, K., Breimhorst, M., Noachtar, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Latency+of+interictal+epileptiform+discharges+in+long-term+EEG+recordings+in+epilepsy+patients.">Latency of interictal epileptiform discharges in long-term EEG recordings in epilepsy patients.</a> Seizure. 29, 20-25 (2015).</li><li>Staba, R. J., Stead, M., Worrell, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+biomarkers+of+epilepsy.">Electrophysiological biomarkers of epilepsy.</a> Neurotherapeutics. 11 (2), 334-346 (2014).</li><li>Nimmrich, V., Draguhn, A., Axmacher, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+Network+Oscillations+in+Neurodegenerative+Diseases.">Neuronal Network Oscillations in Neurodegenerative Diseases.</a> Neuromolecular Med. 17 (3), 270-284 (2015).</li><li>Zayachkivsky, A., Lehmkuhle, M. J., Dudek, F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+Continuous+EEG+Monitoring+in+Small+Rodent+Models+of+Human+Disease+Using+the+Epoch+Wireless+Transmitter+System.">Long-term Continuous EEG Monitoring in Small Rodent Models of Human Disease Using the Epoch Wireless Transmitter System.</a> J Vis Exp. (101), e52554 (2015).</li><li>Zayachkivsky, A., Lehmkuhle, M. J., Fisher, J. H., Ekstrand, J. J., Dudek, F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recording+EEG+in+immature+rats+with+a+novel+miniature+telemetry+system.">Recording EEG in immature rats with a novel miniature telemetry system.</a> J Neurophysiol. 109 (3), 900-911 (2013).</li><li>Dzhala, V. I., 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=NKCC1+transporter+facilitates+seizures+in+the+developing+brain.">NKCC1 transporter facilitates seizures in the developing brain.</a> Nat Med. 11 (11), 1205-1213 (2005).</li><li>Tucker, A. M., Aquilina, K., Chakkarapani, E., Hobbs, C. 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作者宣称没有利益冲突。

在这里，我们报告手术和记录程序，通过有线方法自由移动新生大鼠幼仔获得脑电图（ 图3 ）。已经表明P7-P12大鼠的发育年龄对应于全新的人类新生儿10,11 。在这个年龄组的大鼠幼仔工作时，在技术上难以获得高质量的脑电记录数据。此外，它需要专门的处理和很好的保养8 。 以前在新生大鼠体内检测体内 EEG记录的研究已经在P12或更老的动物12中使用有线记录解决方案进行了，但是这些方法是非常昂贵的。通过依靠简单的电极，我们已经能够制造一个电极（ 图1 ），将实验成本降低到低至1美元，同时仍然给予高密度脑电图录音。这种方法的手术植入可能需要10分钟，这取决于手术的复杂性。 该协议中的关键步骤是电极的注入。应该小心点为了保持电极与头骨的恒定接触，以下步骤是重要的。首先，用灭菌棉签去除颅骨上的组织碎屑，因为它可能会使电极与头骨分开。其次，使用氰基丙烯酸酯适当地附着电极与颅骨。第三，保持牙膏粘稠度。最好在两层涂抹牙科水泥;第一层应该有点密集以覆盖头骨，而第二层应该有点流氓覆盖第一层的角落。用移液管尖端轻轻敲打牙科水泥，将其固定下来，使粘结剂粘结牢固。如果它太水了，它可能在电极下面建立一个绝缘层ode，如果它太密集，牙科水泥将由于自身的重量而容易从头骨脱落。在整个手术期间保持小鼠的体温在37°C生存也很重要。 该程序的应用限于在P8和P15年龄之间的新生大鼠幼仔中记录脑电图，因为生长的幼仔可以从其头骨中去除牙科水泥和电极。在进一步的应用方面，该方法也可以用于记录成年大鼠和小鼠在稍微修改后的电活动。此外，该方法很好地适用于各种行为实验，例如高架加迷宫。

已经确定，需要神经元之间的连续通信来获得正常的脑功能。神经元间通讯主要发生在突触处，其中一个神经元的信息被传送到第二个神经元。这种突触传播由两种类型的专用结构布置介导:电或化学突触1 。电生理学是捕获在神经间通讯期间产生的控制整个身体功能和行为的电位的领域。脑电是许多电生理技术中最常用的方法。 脑电图是用于检测由内部或外部刺激产生的电信号变化的技术。此外，它是临床诊断和各种神经疾病如癫痫，帕金森病和阿尔茨海默氏症患者预后的重要测试e，以及药理和毒理剂的作用3 。一般来说，癫痫患者在脑内表现出过度兴奋性和功能性连接受损;这些被概括为间质性癫痫样放电（IEDs），可以用尖锐的瞬时尖峰的形式记录脑电图。尖锐的波浪尖波复合物或多峰4 。癫痫大脑的主要特征是癫痫发作的自发性发作，可以从头皮或脑实质记录，以定位癫痫发作的脑区5 。此外，EEG对神经变性疾病如阿尔茨海默氏病（AD）也具有非常重要的意义。研究表明，AD患者的EEG记录和受损振荡网络的改变是常见的。然而，我们对神经退行性疾病网络振荡的病理生理学知识令人惊讶的是不完整，需要进一步探索6 。 在这个协议中，我们设计了一个简单的电极，可以记录EEG以了解正常和病理大脑的电气通信。这种方法的手术植入比其他可用的手术便宜7 。此外，该方法可用于记录更长时间内的高质量和稳定的EEG信号（ 即每天2-4小时1周）。此外，我们使用更轻的电极（重约26毫克），使动物的行为更自然8 。该方法广泛适用于需要放大器和数字化仪的新生大鼠幼仔的脑电图研究，通常用于电生理实验室，不需要任何额外的装置。

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recording eeg in freely moving neonatal rats using a novel method

脑电图是检测大脑电活动的有用方法。此外，它是广泛使用的各种神经病症的诊断工具，如癫痫和神经退行性疾病。然而，在技术上难以在新生儿获得脑电记录，因为它需要专门的处理和很好的照顾。在这里，我们提出了一种在新生大鼠幼仔（P8-P15）中记录脑电图的新方法。我们设计了一个使用计算机针座的简单可靠的电极;它可以很容易地植入大鼠的颅骨，以在正常和癫痫大脑中记录高质量的脑电信号。给予小鼠腹膜内（ip）注射神经毒素红藻氨酸（KA）以诱导癫痫发作。在这个程序中进行的手术植入比其他新生儿的EEG手术便宜。该方法允许一个记录高质量和稳定的EEG信号超过1周。此外，该程序也可以应用于成人rats和小鼠研究癫痫或其他神经系统疾病。

<html>如果上述外科手术正确进行，则可以成功地进行一次大鼠新生儿小脑脑电图记录。 KA注射后10分钟，出现不规则运动和搔抓，震颤和失去平衡的形式的行为体征。图2显示了代表性的原始脑电图痕迹和间质，炎症反应和发作性扩张痕迹。复发性间歇和发作性EEG放电模式在KA注射后开始15-60分钟（30±5.2分钟;平均值±SEM）（ 图2A和2B ）。 图2D和2E表示最常见类型的强直阵挛性发作。发炎持续时间持续15.2±0.9s（ 图2D ）。代表性的结果表明，强直性放电之后是阵挛性爆发（ 图2B，2D和2E）。 <html> e_content"fo:keep-together.within-page ="1"&gt; <img alt="图1" src="/files/ftp_upload/55489/55489fig1.jpg" /> 图1:手术植入方案的例证。 （ A ）微小的电脑引脚。 （ B ）母针（左）和公针（右）。比例尺用于测量母针的长度。 （ C ）连接的母头和针头。 （ D ）暴露的头骨。第1号提供参考地点，第2号呈现了伯尔贾，第3和第4个现有录音场所，第5号呈现了Lambda。 （ E ）电极植入。 1号是参考电极，No.3和4存在记录电极。 （ F ）恢复大鼠幼仔（P9）。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55489/55489fig1large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> d / 55489 / 55489fig2.jpg"/&gt; 图2:KA引起的新生儿癫痫发作。 （ A ）来自注射KA（2mg / kg，ip）的P10大鼠的代表性EEG迹线显示出复发性癫痫突发放电。 （ B ）从A框（ C ， D和E ）中的单个簇状癫痫放电的扩展视图。从B框中扩展出的典型的间质，炎症和惊恐曲线。（ F ）总结数据显示KA注射前后的平均EEG功率（1分钟时间窗）（平均值±SEM）。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55489/55489fig2large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> <img alt="图3" src="/files/ftp_upload/55489/55489fig3.jpg" /> 图3:脑电记录设置。代表追踪记录是sho在自由移动的新生大鼠小狗。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55489/55489fig3large.jpg" target="_blank">请点击此处查看此图的较大版本。</a>

Watch this Scientific Journal Video about Recording EEG in Freely Moving Neonatal Rats Using a Novel Method at JoVE.com

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

在这里，我们介绍一种设计用于记录脑电图（EEG）在自由移动的新生儿癫痫幼仔中的新技术，并描述其程序，特征和应用。这种方法允许一个记录EEG超过1周。

使用新颖的方法在自由移动的新生大鼠中记录脑电图

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such ...

recording-eeg-in-freely-moving-neonatal-rats-using-a-novel-method

Nanyang Technological University

Research

JoVE Journal

Neuroscience

13.1K 次观看.  South China Normal University. 在这里，我们介绍一种设计用于记录脑电图（EEG）在自由移动的新生儿癫痫幼仔中的新技术，并描述其程序，特征和应用。这种方法允许一个记录EEG超过1周。 

视频: Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

Novel Apparatus and Method for Drug Reinforcement

Animal models of reinforcement have proven to be useful for understanding the neurobiological mechanisms underlying drug addiction. Operant drug self-administration and conditioned place preference (CPP) procedures are expansively used in animal research to model various components of drug reinforcement, consumption, and addiction in humans. For this study, we used a novel approach to studying drug reinforcement in rats by combining traditional CPP and self-administration methodologies. We assembled an apparatus using two Med Associate operant chambers, sensory stimuli, and a Plexiglas-constructed neutral zone. These modifications allowed our experiments to encompass motivational aspects of drug intake through self-administration and drug-free assessment of drug/cue conditioning strength with the CPP test. In our experiments, rats self-administered cocaine (0.75 mg/kg/inj, i.v.) during either four (e.g., the "short-term") or eight (e.g., the "long-term") alternating-day sessions in an operant environment containing distinctive sensory cues (e.g., olfactory and visual). On the alternate days, in the other (differently-cued) operant environment, saline was available for self-infusion (0.1 ml, i.v.). Twenty-four hours after the last self-administration/cue-pairing session, a CPP test was conducted. Consistent with typical CPP findings, there was a significant preference for the chamber associated with cocaine self-administration. In addition, in animals undergoing the long-term experiment, a significant positive correlation between CPP magnitude and the number of cocaine-reinforced lever responses. In conclusion, this apparatus and approach is time and cost effective, can be used to examine a wide array of topics pertaining to drug abuse, and provides more flexibility in experimental design than CPP or self-administration methods alone.

Operant drug self-administration and conditioned place preference (CPP) procedures are expansively used in research to model various components of drug reinforcement, consumption, and addiction in humans. In this report, we combined traditional CPP and self-administration methods as a novel approach to studying drug reinforcement and addiction in rats.

新型仪器和药物加固方法

Animal models of reinforcement have proven to be useful for understanding the neurobiological mechanisms underlying drug addiction.  Operant drug ...

科研

神经科学

High-density EEG Recordings of the Freely Moving Mice using Polyimide-based Microelectrode

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a noninvasive brain monitoring tool in cognitive neuroscience as well as a diagnostic tool for epilepsy and sleep disorders in neurology. However, the underlying mechanism of EEG rhythm generation is still under the veil. Recently introduced polyimide-based microelectrode (PBM-array) for high resolution mouse EEG1 is one of the trials to answer the neurophysiological questions on EEG signals based on a rich genetic resource that the mouse model contains for the analysis of complex EEG generation process. This application of nanofabricated PBM-array to mouse skull is an efficient tool for collecting large-scale brain activity of transgenic mice and accommodates to identify the neural correlates to certain EEG rhythms in conjunction with behavior. However its ultra-thin thickness and bifurcated structure cause a trouble in handling and implantation of PBM-array. In the presented video, the preparation and surgery steps for the implantation of PBM-array on a mouse skull are described step by step. Handling and surgery tips to help researchers succeed in implantation are also provided.

In this article, we described the surgery procedure and handling tips for implantation of ultra-thin polyimide-based microelectrode array (PBM-array) on the mouse skull for acquisition of high-density encephalography (EEG) in a mouse model.

高密度的自由移动，使用基于聚酰亚胺的微电极的小鼠的脑电图记录

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a ...

A General Method for Evaluating Incubation of Sucrose Craving in Rats

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and relapse to food taking 1. Food craving conceptualized in this way is akin to drug craving in response to drug-paired cues. A rich literature has been developed around understanding the behavioral and neurobiological determinants of drug craving; we and others have been focusing recently on translating techniques from basic addiction research to better understand addiction-like behaviors related to food 2-4.
As done in previous studies of drug craving, we examine sucrose craving behavior by utilizing a rat model of relapse. In this model, rats self-administer either drug or food in sessions over several days. In a session, lever responding delivers the reward along with a tone+light stimulus. Craving behavior is then operationally defined as responding in a subsequent session where the reward is not available. Rats will reliably respond for the tone+light stimulus, likely due to its acquired conditioned reinforcing properties 5. This behavior is sometimes referred to as sucrose seeking or cue reactivity. In the present discussion we will use the term "sucrose craving" to subsume both of these constructs.
In the past decade, we have focused on how the length of time following reward self-administration influences reward craving. Interestingly, rats increase responding for the reward-paired cue over the course of several weeks of a period of forced-abstinence. This "incubation of craving" is observed in rats that have self-administered either food or drugs of abuse 4,6. This time-dependent increase in craving we have identified in the animal model may have great potential relevance to human drug and food addiction behaviors.
Here we present a protocol for assessing incubation of sucrose craving in rats. Variants of the procedure will be indicated where craving is assessed as responding for a discrete sucrose-paired cue following extinction of lever pressing within the sucrose self-administration context (Extinction without cues) or as responding for sucrose-paired cues in a general extinction context (Extinction with cues).

Responding for food or drug-paired cues increases over the course of a period of abstinence and this may relate to an increased susceptibility to relapse behaviors. Here we detail a procedure for evaluating this "incubation of craving" in rats that have self-administered sucrose.

评估大鼠蔗糖渴求育成的一个通用方法

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and ...

Recording Single Neurons' Action Potentials from Freely Moving Pigeons Across Three Stages of Learning

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed single-neuron activity while animals are acquiring an operantly conditioned response, or when that response is extinguished. But even in these cases, observation periods usually encompass only a single stage of learning, i.e. acquisition or extinction, but not both (exceptions include protocols employing reversal learning; see Bingman&#160;et al.1 for an example). However, acquisition and extinction entail different learning mechanisms and are therefore expected to be accompanied by different types and/or loci of neural plasticity.
Accordingly, we developed a behavioral paradigm which institutes three stages of learning in a single behavioral session and which is well suited for the simultaneous recording of single neurons&#39; action potentials. Animals are trained on a single-interval forced choice task which requires mapping each of two possible choice responses to the presentation of different novel visual stimuli (acquisition). After having reached a predefined performance criterion, one of the two choice responses is no longer reinforced (extinction). Following a certain decrement in performance level, correct responses are reinforced again (reacquisition). By using a new set of stimuli in every session, animals can undergo the acquisition-extinction-reacquisition process repeatedly. Because all three stages of learning occur in a single behavioral session, the paradigm is ideal for the simultaneous observation of the spiking output of multiple single neurons. We use pigeons as model systems, but the task can easily be adapted to any other species capable of conditioned discrimination learning.

Learning new stimulus-response associations engages a wide range of neural processes which are ultimately reflected in changing spike output of individual neurons. Here we describe a behavioral protocol allowing for the continuous registration of single-neuron activity while animals acquire, extinguish, and reacquire a conditioned response within a single experimental session.

自由移动鸽子穿越三个阶段学习记录单个神经元&#39;动作电位

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed ...

Extracellular Wire Tetrode Recording in Brain of Freely Walking Insects

Increasing interest in the role of brain activity in insect motor control requires that we be able to monitor neural activity while insects perform natural behavior. We previously developed a technique for implanting tetrode wires into the central complex of cockroach brains that allowed us to record activity from multiple neurons simultaneously while a tethered cockroach turned or altered walking speed. While a major advance, tethered preparations provide access to limited behaviors and often lack feedback processes that occur in freely moving animals. We now present a modified version of that technique that allows us to record from the central complex of freely moving cockroaches as they walk in an arena and deal with barriers by turning, climbing or tunneling. Coupled with high speed video and cluster cutting, we can now relate brain activity to various parameters of the movement of freely behaving insects.

We previously developed a technique for implanting tetrode wires into the central complex of cockroach brains that allows us to monitor activity in individual units of tethered cockroaches. Here we present a modified version of that technique that allows us to also record brain activity in freely moving insects.

外导线四极管记录在行走自如昆虫大脑

Increasing interest in the role of brain activity in insect motor control requires that we be able to monitor neural activity while insects perform ...

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees (Bombus terrestris)

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from flowering plants. Only a few studies have described the gustatory responses of bees toward toxins in food, and these experiments have mainly used the proboscis extension response on restrained honey bees. Here, a new behavioral assay is presented for measuring the feeding responses of freely-moving, individual worker bumble bees to nutrients and toxins. This assay measures the amount of solution ingested by each bumble bee and identifies how tastants in food influence the microstructure of the feeding behavior.
The solutions are presented in a microcapillary tube to individual bumble bees that have been previously starved for 2-4 hr. The behavior is captured on digital video. The fine structure of the feeding behavior is analyzed by continuously scoring the position of the proboscis (mouthparts) from video recordings using event logging software. The position of the proboscis is defined by three different behavioral categories: (1) proboscis is extended and in contact with the solution, (2) proboscis is extended but not in contact with the solution and (3) proboscis is stowed under the head. Furthermore the speed of the proboscis retracting away from the solution is also estimated.
In the present assay the volume of solution consumed, the number of feeding bouts, the duration of the feeding bouts and the speed of the proboscis retraction after the first contact is used to evaluate the phagostimulatory or the deterrent activity of the compounds tested.
This new taste assay will allow researchers to measure how compounds found in nectar influence the feeding behavior of bees and will also be useful to pollination biologists, toxicologists and neuroethologists studying the bumble bee&#39;s taste system.

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees Bombus terrestris

A novel behavioral assay is described for investigating the short term gustatory responses of the mouthparts of freely-moving bumble bees (Bombus terrestris) toward nutrients and toxins in solution.

一种新的行为分析调查个人的味觉，可自由移动的熊蜂（<em&gt;欧洲熊蜂</em&gt;）

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from ...

Noninvasive EEG Recordings from Freely Moving Piglets

The method allows the recording of high-quality electroencephalograms (EEGs) from freely moving piglets directly in the pigpen. We use a one-channel telemetric electroencephalography system in combination with standard self-adhesive hydrogel electrodes. The piglets are calmed down without the use of sedatives. After their release into the pigpen, the piglets behave normally&#8212;they drink and sleep in the same cycle as their siblings. Their sleep phases are used for the EEG recordings.

Here, we present a protocol to record telemetric electroencephalograms (EEGs) from freely moving piglets directly in the pigpen without the use of a sedative, making it possible to record typical EEG patterns during non-REM sleep, like spindle bursts.

自由移动仔猪的无创脑电图记录

The method allows the recording of high-quality electroencephalograms (EEGs) from freely moving piglets directly in the pigpen. We use a one-channel ...

Microdialysis of Excitatory Amino Acids During EEG Recordings in Freely Moving Rats

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain interstitial space with the behavior and/or with the specific outcome of a pathology (e.g., seizures for epilepsy). When studying epilepsy, the microdialysis technique is often combined with short-term or even long-term video-electroencephalography (EEG) monitoring to assess spontaneous seizure frequency, severity, progression and clustering. The combined microdialysis-EEG is based on the use of several methods and instruments. Here, we performed in vivo microdialysis and continuous video-EEG recording to monitor glutamate and aspartate outflow over time, in different phases of the natural history of epilepsy in a rat model. This combined approach allows the pairing of changes in the neurotransmitter release with specific stages of the disease development and progression. The amino acid concentration in the dialysate was determined by liquid chromatography. Here, we describe the methods and outline the principal precautionary measures one should take during in vivo microdialysis-EEG, with particular attention to the stereotaxic surgery, basal and high potassium stimulation during microdialysis, depth electrode EEG recording and high-performance liquid chromatography analysis of aspartate and glutamate in the dialysate. This approach may be adapted to test a variety of drug or disease induced changes of the physiological concentrations of aspartate and glutamate in the brain. Depending on the availability of an appropriate analytical assay, it may be further used to test different soluble molecules when employing EEG recording at the same time.

Here, we describe a method for in vivo microdialysis to analyze aspartate and glutamate release in the ventral hippocampus of epileptic and non-epileptic rats, in combination with EEG recordings. Extracellular concentrations of aspartate and glutamate may be correlated with the different phases of the disease.

自由运动大鼠脑电图记录过程中兴奋氨基酸的微透析

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain ...

Hybrid Microdrive System with Recoverable Opto-Silicon Probe and Tetrode for Dual-Site High Density Recording in Freely Moving Mice

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, conventional microdrive designs often only allow use of one type of electrode to record from single or multiple regions, limiting the yield of single-unit or depth profile recordings. It also often limits the ability to combine electrode recordings with optogenetic tools to target pathway and/or cell type specific activity. Presented here is a hybrid microdrive array for freely moving mice to optimize yield and a description of its fabrication and reuse of the microdrive array. The current design employs nine tetrodes and one opto-silicon probe implanted in two different brain areas simultaneously in freely moving mice. The tetrodes and the opto-silicon probe are independently adjustable along the dorsoventral axis in the brain to maximize the yield of unit and oscillatory activities. This microdrive array also incorporates a set-up for light, mediating optogenetic manipulation to investigate the regional- or cell type-specific responses and functions of long-range neural circuits. In addition, the opto-silicon probe can be safely recovered and reused after each experiment. Because the microdrive array consists of 3D-printed parts, the design of microdrives can be easily modified to accommodate various settings. First described is the design of the microdrive array and how to attach the optical fiber to a silicon probe for optogenetics experiments, followed by fabrication of the tetrode bundle and implantation of the array into a mouse brain. The recording of local field potentials and unit spiking combined with optogenetic stimulation also demonstrate feasibility of the microdrive array system in freely moving mice.

This protocol describes the construction of a hybrid microdrive array that allows implantation of nine independently adjustable tetrodes and one adjustable opto-silicon probe in two brain regions in freely moving mice. Also demonstrated is a method for safely recovering and reusing the opto-silicon probe for multiple purposes.

混合微驱系统,带可回收的光电探头和Tetrode,用于自由移动小鼠的双位点高密度记录

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, ...

Multi-Fiber Photometry to Record Neural Activity in Freely-Moving Animals

Recording the activity of a group of neurons in a freely-moving animal is a challenging undertaking. Moreover, as the brain is dissected into smaller and smaller functional subgroups, it becomes paramount to record from projections and/or genetically-defined subpopulations of neurons. Fiber photometry is an accessible and powerful approach that can overcome these challenges. By combining optical and genetic methodologies, neural activity can be measured in deep brain structures by expressing genetically-encoded calcium indicators, which translate neural activity into an optical signal that can be easily measured. The current protocol details the components of a multi-fiber photometry system, how to access deep brain structures to deliver and collect light, a method to account for motion artifacts, and how to process and analyze fluorescent signals. The protocol details experimental considerations when performing single and dual color imaging, from either single or multiple implanted optic fibers.

This protocol details how to implement and perform multi-fiber photometry recordings, how to correct for calcium-independent artifacts, and important considerations for dual-color photometry imaging.

多纤维光测量记录自由移动动物的神经活动

Recording the activity of a group of neurons in a freely-moving animal is a challenging undertaking. Moreover, as the brain is dissected into smaller and ...