我们要感谢 Scheu 的机会, 在猪圈 Hofgut Neumühle 进行我们的研究。

所有程序均经当地道德委员会批准 (#23177 07/G10-1-010/G 15-15-011), 并遵循欧洲和德国国家条例 (欧洲共同体理事会指令, 86/609/ECC;Tierschutzgesetz)。
所有动物程序都是按照古腾堡的医疗中心-大学美因法动物保育委员会的规定进行的。
1. 设置
<ol><li>在实验之前, 检查任何线路噪音, 并找到一个适当的位置, 设置和天线。线噪声是可见的60或50赫兹正弦波。 注意: 天线的位置, 特别是发射机和接收器之间的距离取决于系统的传输强度。这里使用的系统是可调整的。它被调整了到一个相对地低力量, 与大约 3 m 传输范围。此外, 猪圈中的金属栅栏可以抑制信号并引起干扰。在这种情况下, 有必要将天线放在金属笼内。</li>
	<li>使用电缆鼓提供线路电源设置。将笔记本电脑、接收器单元和模拟数字转换器 (如有必要) 连接到正在使用的特定遥测系统。 注: 这里使用的遥测系统向接收器发送数字数据。这可能与其他系统不同。</li>
	<li>将电极、粘合剂、Q 尖和湿巾以及混合块放在一个单独的桌子上。</li>
	<li>用短电缆准备电极。要做到这一点, 切电极和焊料, 其长度尽可能短, 取决于动物的大小。电缆必须足够长的时间将所需的记录位置与遥测脑电图单元连接, 传输数据。太长的电缆必须退缩和覆盖与皮肤胶粘剂硅胶弹性体。更长的电缆必须退缩, 使硅胶补丁更大, 更重。</li>
</ol>2. 小猪
<ol><li>抓一只小猪, 抓住它的腿或胸部。保持它, 并注意任何大便或排尿。</li>
	<li>如有必要, 用数字标记小猪。</li>
	<li>把小猪裹在毛巾里。小猪会冷静下来的。注意使小猪过热。</li>
	<li>用一只手握住小猪的身体或前臂。用另一只手握住鼻子。注意使小猪过热, 并确保它是自由呼吸正常。</li>
</ol>3. 电极
<ol><li>有第二个人连接电极。</li>
	<li>用水或乙醇清洁皮肤。如有必要, 剃掉头。</li>
	<li>用磨料脑电图凝胶和 Q 尖清除任何死皮细胞。以后去掉磨料凝胶。或者, 使用砂纸。</li>
	<li>固定在所需位置的自粘电极。将接地电极置于小脑 (耳朵之间) 和鼻子上的参考电极之上。将录音电极放在所需位置。 注: 在这种情况下, 执行单极录音, 因为参考被放置在一个中性位置 (鼻子)。目前尚无标准的仔猪可供使用。在这里, 在右脑半球使用了一个顶叶记录位置 (眼睛和耳朵之间)。</li>
	<li>将电缆连接到遥测设备。打开设备。根据所使用的遥测系统, 这可能是一个磁开关或射频唤醒信号。</li>
	<li>覆盖遥测装置和所有电缆以及所有电极与双组分皮肤胶粘剂硅橡胶 (参见材料表)。通过混合相同数量的两个组分, 固化时间将在1分钟的范围内, 眼睛和睫毛不应该覆盖橡胶。</li>
	<li>等到硅橡胶完全固化。</li>
	<li>把小猪放回猪圈。</li>
	<li>观察小猪, 看看它是否在较长的时间内显示出不适的迹象 (几分钟)。</li>
</ol>4. 测量
<ol><li>等待, 直到小猪恢复, 并开始同步其行为与它的同胞 (喂养, 玩耍, 睡眠), 通常在三十年代后 (图 1)。</li>
	<li>如果需要, 等待睡眠阶段。记录时间取决于具体的科学问题。这里使用了10分钟的录音会话。</li>
	<li>如果遥测单元的覆盖范围超过2其他仔猪, 信号可能过低, 接收器。如果他们睡在上面, 轻轻地把小猪推开。注意母猪;它可能会有积极的反应。</li>
	<li>用数据采集软件开始录音 (见材料表)。</li>
</ol>5. 完成
<ol><li>录音之后 (通常几个小时), 再次捕捉小猪, 正如在步骤2中所描述的那样。注意母猪;它可能会有积极的反应。</li>
	<li>轻轻地举起硅橡胶在一边。然后, 除去含有电极和遥测装置的硅橡胶的整个补丁。小心小猪的眼睛。</li>
	<li>把小猪放回猪圈。</li>
</ol>

<ol>
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作者没有什么可透露的。

该协议的一个关键步骤是适当的皮肤接触与电极, 特别是接地电极, 以实现稳定的录音, 低噪音。此外, 由于仔猪非常敏捷, 所以必须用硅橡胶覆盖整个系统, 以保护电极和遥测装置。此外, 如果实验是在一个稳定的板条地板, 小心与小设备或连接器。
如果不充分掌握自粘性水凝胶电极, 尽量去除磨料奶油尽可能。另一种去除外层死皮层的方法是使用砂纸。酒精的预处理不是强制性的。肮脏的皮肤也可以用清水清洗。请勿使用石膏条代替车身双硅胶固定遥测系统。石膏带会引起皮肤发炎和大量的不适症状。在这种情况下, 小猪很快就能摆脱这个系统。
该方法的局限性是水凝胶电极的稳定性。他们可能在几个小时后干燥, 导致记录质量的损失。此外, 与所有脑电图技术一样, 强烈的运动通常与记录跟踪的肌肉工件相关 (如图 4所示)。另外, 天线的良好位置对于减少可能因接收器信号差而产生的工件很重要。金属栅栏也会引起高频干扰, 导致数据序列丢失。解决这个问题的办法是把天线放在金属栅栏的最佳位置。该天线的最佳位置只能通过试验和误差在现场进行评估。母猪和其他仔猪不会导致人工制品, 因为服用该系统的仔猪通常不允许其他仔猪操纵头部。这可能与其他物种不同。
该方法对于神经科学中的平移方法尤为重要。有了这种技术, 就有可能在没有镇静的情况下使用无创记录。由此产生的脑电图记录与临床设置非常相似。该技术可能开辟新的可能性, 以表征早产儿脑电图成熟的手段, 动物模型系统。除此之外, 许多神经科学研究的问题, 特别是关于皮质领域的潜力, 也可能由这种无创技术检查。因此, 该方法有可能减少神经科学领域动物实验的数量和严重性。
未来, 有超过一个电极的录音计划。先决条件是电极的小型化。此外, 电极的长期稳定性是未来创新的一个问题。

仔猪是神经科学1的新兴模型系统。为了加强翻译研究, 我们发明了一种从无节制仔猪2 (图 1和图 2) 中记录无侵袭性、临床脑电图的方法。对脑电图记录的翻译使用的两个先决条件, 关于与皮质成熟有关的脑电图模式, 是一种非侵入性的方法, 可与临床设置相媲美, 也可以节制镇静剂或麻醉。单通道遥测系统3与自粘电极结合, 可固定在5分钟左右. 此后, 仔猪将迅速从处理过程中恢复, 并将其喂养和睡眠行为与其他小猪和母猪。
虽然已经有尝试使用非侵入性脑电图记录从镇静动物4, 多数脑电图研究从动物是以侵入的方法进行。这些方法对植入电极5、6周围的炎症过程有副作用, 在大多数情况下, 由于植入脑电图系统的外部成分, 它们需要动物的社会分离。因此, 将这些数据翻译到临床背景是困难的。对翻译方法的需要越来越清楚, 因为它仍然不知道如何在早期皮质发育期间的 "正常" 大脑成熟是由临床, 非侵入性脑电图7表示。这一知识缺口是由8早产儿脑电图记录所带来的技术挑战引起的。在动物模型系统中, 早期皮质发育的模式更容易获得, 因为大多数动物与人类皮质发育相比, 出生时有 "早产儿"9。除了2种动物皮质发育的保守模式外, 最近还发现早产儿的脑电图记录也可以预测10、11以后的个体临床结局。这里描述的方法对于发展神经科学的转化方面特别有用。

<table border="0" cellpadding="0" cellspacing="0" width="867">
	<tbody>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Disposable adhesive 
			surface silver/silver chloride electrodes</td>
			<td style="width:309px;">Spes 
			Medica S.r.l., Genova, Italy</td>
			<td style="width:119px;"></td>
			<td style="width:223px;">Self adhesive hydrogel electrode</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Abralyt HiCl</td>
			<td style="width:309px;">Easycap GmbH</td>
			<td style="width:119px;"></td>
			<td>Abrasive cream</td>
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			<td height="42" style="height:42px;width:216px;">Body Double fast</td>
			<td style="width:309px;">Smooth On Inc.</td>
			<td style="width:119px;"></td>
			<td>Skin adhesive silicone</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Telemetry system</td>
			<td style="width:309px;"></td>
			<td style="width:119px;"></td>
			<td>Internal development</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Picolog 1216</td>
			<td style="width:309px;">Pico Technology</td>
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			<td>AD converter</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Laptop</td>
			<td style="width:309px;">Panasonic</td>
			<td style="width:119px;"></td>
			<td>Rugged laptop</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Receiver</td>
			<td style="width:309px;"></td>
			<td style="width:119px;"></td>
			<td>Internal development</td>
		</tr>
	</tbody>
</table>

noninvasive eeg recordings from freely moving piglets

该方法允许在猪圈中直接从自由移动的仔猪中记录高质量的脑电图 (脑电图)。采用单通道遥测脑电图系统与标准自粘水凝胶电极结合使用。小猪在没有镇静剂的情况下平静下来。在释放到猪圈后, 小猪的行为正常--他们在同兄弟姐妹一样的循环中喝酒睡觉。他们的睡眠阶段用于脑电图录音。

我们能够记录与非 REM 睡眠相关的典型脑电图模式, 如主轴爆裂或三角洲刷子, 从自由移动的仔猪 (图 1和图 2)。我们主要对非 REM 睡眠期间的代表性模式感兴趣, 但也记录了一个非常低振幅的 REM 睡眠12的阶段 (图 3)。13种动物的生理和 REM 睡眠量各不相同。短 REM 阶段在几分钟的范围是典型的为猪14。在喂养 (哺乳) 期间也可获得良好的录音质量 (图 4)。对于玩耍行为, 肌肉活动会导致肌肉制品;然而, 过滤器的设计, 以提取脑电图带。进一步可能的分析工具是计算功率谱密度或分析工具聚焦于网络活动, 例如, 相位振幅耦合。这些分析工具也可以用于单声道脑电图记录。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/58226/58226fig1.jpg"> 图 1: 一只沉睡的小猪与遥测脑电图系统.<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58226/58226fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/58226/58226fig2.jpg"> 图 2: 在一个自由移动的小猪的睡眠阶段记录的典型脑电图模式, 在它的兄弟姐妹旁边休息.四事件由框突出显示。从左到右, 第一个框显示肌肉活动引起的典型模式;例如, 睡眠中的短抽搐, 其特点是高剂量的快速伽玛活动 (超过80赫兹) 和爆裂样的外观。第二个框显示一个三角洲画笔类似的情节, 其特点是三角洲活动与叠加活动在θ和 alpha 范围。第三和第四盒显示伽玛爆发的短集 (睡眠纺锤状事件), 其特征是α、β和伽玛波段的频率分量。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58226/58226fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/58226/58226fig3.jpg"> 图 3: 睡眠期间的 REM 阶段.第一个和第二个框显示慢波睡眠阶段。第三个框表示大约二十年代的低振幅脑电图阶段在期间。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58226/58226fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/58226/58226fig4.jpg"> 图 4: 一个10分钟的脑电图记录一只饮用的小猪 (哺乳) 的踪迹.100s 后的低谷是由于发射机和脑电图记录系统的接收器之间的无线电通信的短的损失。之后, 有一些肌肉或运动的工件。肌肉制品的特点是非常高的三角洲带振幅。相比之下, EEG 活动的特点是从慢到快波的振幅功率逐渐下降 (见, 例如, 200 和 250s)。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58226/58226fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Noninvasive EEG Recordings from Freely Moving Piglets at JoVE.com

Noninvasive EEG Recordings from Freely Moving Piglets

在这里, 我们提出了一个协议, 记录遥测脑电图 (脑电图) 从自由移动仔猪直接在猪圈没有使用镇静剂, 使其能够记录典型的脑电图模式在非 REM 睡眠, 如纺锤爆发。

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 ...

noninvasive-eeg-recordings-from-freely-moving-piglets

Nanyang Technological University

Research

JoVE Journal

Neuroscience

7.2K 次观看.  Free University Berlin. 在这里, 我们提出了一个协议, 记录遥测脑电图 (脑电图) 从自由移动仔猪直接在猪圈没有使用镇静剂, 使其能够记录典型的脑电图模式在非 REM 睡眠, 如纺锤爆发。

视频: Noninvasive EEG Recordings from Freely Moving Piglets

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 ...

科研

神经科学

Patch Clamp Recordings from Embryonic Zebrafish Mauthner Cells

Mauthner cells (M-cells) are large reticulospinal neurons located in the hindbrain of teleost fish. They are key neurons involved in a characteristic behavior known as the C-start or escape response that occurs when the organism perceives a threat. The M-cell has been extensively studied in adult goldfish where it has been shown to receive a wide range of excitatory, inhibitory and neuromodulatory signals1. We have been examining M-cell activity in embryonic zebrafish in order to study aspects of synaptic development in a vertebrate preparation. In the late 1990s Ali and colleagues developed a preparation for patch clamp recording from M-cells in zebrafish embryos, in which the CNS was largely intact2,3,4. The objective at that time was to record synaptic activity from hindbrain neurons, spinal cord neurons and trunk skeletal muscle while maintaining functional synaptic connections within an intact brain-spinal cord preparation. This preparation is still used in our laboratory today. To examine the mechanisms underlying developmental synaptic plasticity, we record excitatory (AMPA and NMDA-mediated)5,6 and inhibitory (GABA and glycine) synaptic currents from developing M-cells. Importantly, this unique preparation allows us to return to the same cell (M-cell) from preparation to preparation to carefully examine synaptic plasticity and neuro-development in an embryonic organism. The benefits provided by this preparation include 1) intact, functional synaptic connections onto the M-cell, 2) relatively inexpensive preparations, 3) a large supply of readily available embryos 4) the ability to return to the same cell type (i.e. M-cell) in every preparation, so that synaptic development at the level of an individual cell can be examined from fish to fish, and 5) imaging of whole preparations due to the transparent nature of the embryos.

We have developed an intact brain-spinal cord preparation to record and monitor electrical activity via patch clamp recording from the Mauthner neurons and other reticulospinal cells in zebrafish embryos. Thus, we are able to record excitatory and inhibitory synaptic currents, voltage-gated channel activity and action potentials from key neurons in a developing embryo.

从胚胎斑马鱼的毛特纳细胞膜片钳记录

Mauthner cells (M-cells) are large  reticulospinal neurons located in the hindbrain of teleost fish. They are key  neurons involved in a characteristic ...

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 ...

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 ...

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

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 as epilepsy and neurodegenerative disorders. However, it is technically difficult to obtain EEG recordings in neonates as it requires specialized handling and great care. Here, we present a novel method to record EEG in neonatal rat pups (P8-P15). We designed a simple and reliable electrode using computer pin loci; it can be easily implanted into the skull of a rat pup to record high-quality EEG signals in the normal and epileptic brain. Pups were given an intraperitoneal (i.p.) injection of the neurotoxin kainic acid (KA) to induce epileptic seizures. The surgical implantation performed in this procedure is less expensive than other EEG procedures for neonates. This method allows one to record high-quality and stable EEG signals for more than 1 week. Furthermore, this procedure can also be applied to adult rats and mice to study epilepsy or other neurological disorders.

Here, we introduce a novel technique designed to record electroencephalography (EEG) in freely moving neonatal epileptic pups and describe its procedures, features, and applications. This method allows one to record EEG for more than 1 week.

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 ...

Visual Evoked Potential Recordings in Mice Using a Dry Non-invasive Multi-channel Scalp EEG Sensor

For scalp EEG research environments with laboratory mice, we designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable because of the plunger-spring-barrel structural facet and mechanical strengths resulting from metal materials. The whole process for acquiring the VEP responses in vivo from a mouse consists of four steps: (1) sensor assembly, (2) animal preparation, (3) VEP measurement, and (4) signal processing. This paper presents representative measurements of VEP responses from multiple mice with a submicro-voltage signal resolution and sub-hundred millisecond temporal resolution. Although the proposed method is safer and more convenient compared to other previously reported animal EEG acquiring methods, there are remaining issues including how to enhance the signal-to-noise ratio and how to apply this technique with freely moving animals. The proposed method utilizes easily available resources and shows a repetitive VEP response with a satisfactory signal quality. Therefore, this method could be utilized for longitudinal experimental studies and reliable translational research exploiting non-invasive paradigms.

We designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable. This paper describes the whole process from manufacturing the proposed EEG electrode to signal processing of visual evoked potential (VEP) signals measured on a mouse scalp using a dry non-invasive multi-channel EEG sensor.

干无创多通道头皮脑电传感器在小鼠视觉诱发电位记录中的应用

For scalp EEG research environments with laboratory mice, we designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable ...

Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls body functions and internal organ rhythms when animals are exposed to emotional challenges and changes in their living environments. In general experiments, signals from different organs, such as the brain and the heart, are recorded by independent recording systems that require multiple recording devices and different procedures for processing the data files. This study describes a new method that can simultaneously monitor electrical biosignals, including tens of local field potentials in multiple brain regions, electrocardiograms that represent the cardiac rhythm, electromyograms that represent awake/sleep-related muscle contraction, and breathing signals, in a freely moving rat. The recording configuration of this method is based on a conventional micro-drive array for cortical local field potential recordings in which tens of electrodes are accommodated, and the signals obtained from these electrodes are integrated into a single electrical board mounted on the animal's head. Here, this recording system was improved so that signals from the peripheral organs are also transferred to an electrical interface board. In a single surgery, electrodes are first separately implanted into the appropriate body parts and the target brain areas. The open ends of all of these electrodes are then soldered to individual channels of the electrical board above the animal's head so that all of the signals can be integrated into the single electrical board. Connecting this board to a recording device allows for the collection of all of the signals into a single device, which reduces experimental costs and simplifies data processing, because all data can be handled in the same data file. This technique will aid the understanding of the neurophysiological correlates of the associations between central and peripheral organs.

This study introduces a method for the simultaneous recording of local field potentials in the brain, electrocardiograms, electromyograms, and breathing signals of a freely moving rat. This technique, which reduces experimental costs and simplifies data analysis, will contribute to the understanding of the interactions between the brain and peripheral organs.

自由运动大鼠皮质局部电位、心电图、肌电图和呼吸节律的同步记录

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls ...

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 ...

Long-term Sensory Conflict in Freely Behaving Mice

Long-term sensory conflict protocols are a valuable means of studying motor learning. The presented protocol produces a persistent sensory conflict for experiments aimed at studying long-term learning in mice. By permanently wearing a device fixed on their heads, mice are continuously exposed to a sensory mismatch between visual and vestibular inputs while freely moving in home cages. Therefore, this protocol readily enables the study of the visual system and multisensory interactions over an extended timeframe that would not be accessible otherwise. In addition to lowering the experimental costs of long-term sensory learning in naturally behaving mice, this approach accommodates the combination of in vivo and in vitro experiments. In the reported example, video-oculography is performed to quantify the vestibulo-ocular reflex (VOR) and optokinetic reflex (OKR) before and after learning. Mice exposed to this long-term sensory conflict between visual and vestibular inputs presented a strong VOR gain decrease but exhibited few OKR changes. Detailed steps of device assembly, animal care, and reflex measurements are hereby reported.

The presented protocol produces a persistent sensory conflict for experiments aimed at studying long-term learning. By permanently wearing a fixed device on their heads, mice are continuously exposed to a sensory mismatch between visual and vestibular inputs while freely moving in home cages.

自由生育小鼠的长期感觉冲突

Long-term sensory conflict protocols are a valuable means of studying motor learning. The presented protocol produces a persistent sensory conflict for ...

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, ...