这篇文章得到了中国国家自然科学基金 (31671141) 的支持。

本议定书经当地道德委员会批准。所有参与者和他们的父母都签署了这个实验的知情同意表格。
1. 主题
<ol><li>仅包括15个健康的男性青春期的科目, 年龄从14年到22岁 (意味着标准偏差: 18.3 @ 2.8 年)。 注意: 目前的协议, 分析微状态和欧米茄的复杂性已经发展为健康的对象, 但不限于这个组只。</li>
</ol>2. 脑电图数据记录
<ol><li>让受试者坐在一个安静的, 温控室的舒适的椅子上, 在那里记录脑电图数据。利用蚂蚁脑电图系统在本协议中采集20通道 eeg 数据。</li>
	<li>把帽子放在主题头上。<ol>
<li>在这项研究中, 使用中等大小的帽子, 因为它适合大多数青少年或男性科目。对于儿童, 请测量每个科目的头部周长, 并确定适当大小的帽子。</li>
		<li>将电极直拉在大约50% 的距离 inion 和 nasion 之间, 50% 的距离之间的左和右之间的听觉凹痕。分别将参考电极放在左、右乳突骨上。根据国际10-20 系统, 将其他头皮电极置于标准位置。 注:20 通道的电极系统和 eeg 放大器足以进行脑电图微状态分析和欧米茄复杂度分析。</li>
	</ol>
</li>
	<li>通过电极插入钝针, 填充所有电极与导电凝胶。使用导电凝胶降低阻抗。保持所有电极阻抗低于 10 kiloohms (kΩ)。<ol>
<li>在此期间, 为主题提供一些娱乐 (例如, 放映短片)。如果使用干电极或盐水电极, 则跳过注射导电凝胶的步骤。</li>
	</ol>
</li>
	<li>在录音过程中, 指导受试者闭上眼睛放松, 持续5分钟. 使用数字化软件对脑电图信号进行数字化和记录。使用至少250个样品的取样率. 使用比0.1 和 80 Hz 宽的在线过滤器。 注意: 虽然大多数商用电极放大器系统使用的是有源电极系统, 它可以提高 eeg 信号的信噪比, 但在脑电图记录过程中, 不要将此系统靠近任何电子设备。</li>
</ol>3. 脑电图数据预处理
注: 脑电图数据可以使用各种开源软件或商用软体进行预处理。下面提供的说明是特定于 EEGLAB 的。这只是许多可用选项中的一个来预处理脑电图数据。
<ol><li>将原始脑电图数据导入 eeg 软件 (如 EEGLAB) (文件 |导入数据 |使用 EEGLAB 函数和插件)。 注: 从各种电极放大器系统记录的原始脑电图数据可由 EEGLAB 识别, 如艾吉、蚂蚁、脑视记录仪和 Neuroscan。</li>
	<li>将通道位置文件加载到 EEG 软件中 (编辑 |通道位置)。尽管 eeg 数据和通道名称已导入 EEGLAB, 但为了获得这些电极的空间位置, 请将通道位置文件导入 eeg 软件。</li>
	<li>删除参考电极 (编辑 |选择数据 |在通道范围内选择数据)。在弹出对话框中的 "选择通道范围内的数据" 选项中, 只选择记录电极, 不要选择参考电极, 以便可以删除参考电极。参考电极记录的数据不是 "真实" 的大脑信号, 因为这两个电极分别放在左和右乳突骨上。</li>
	<li>带通过滤器脑电图数据在0.5 和 80 Hz 之间 (工具 |筛选数据 |基本 FIR 过滤器 [新建, 默认])。在弹出对话框中, 为 "频率传递带 (hz)" 的 "下边缘" 选择 5, 并为 "频率传递带 (hz)" 的 "较高边缘" 选择80。然后单击 "确定" 按钮。</li>
	<li>在49和 51 Hz 之间删除带有凹槽过滤器的电源线噪音 (工具 |筛选数据 |基本 FIR 过滤器 [新建, 默认])。在弹出对话框中, 为 "频率传递带 (hz)" 的下边缘选择 49, 然后选择 "频率传递带 (hz) 的更高边缘" 的 51, 然后选择 "凹槽过滤数据而不是传递带" 选项。然后单击 "确定" 按钮。</li>
	<li>使用盲源分离 (BSS) 算法15, 纠正由眼球运动、肌电信号 (肌电图) 或任何其他非生理工件污染的数据部分。对于眼球运动, 点击工具 |使用 AAR 1.3 删除工件 |EOG 去除 |使用 BSS;对于肌电信号, 工具 |使用 AAR 1.3 删除工件 |使用 BSS 去除肌电信号。</li>
	<li>将预处理后的连续脑电图数据分割成世纪, 以2秒为纪元长度。为此, 写 '脑电图 = eeg_regepochs (eeg, ' 复发 ', 2, ' 极限 ', [0 2], ' rmbase ', 南); pop_saveset (EEG)', 然后击中键盘的输入键。一个窗口将弹出, 允许保存分段脑电图数据。</li>
	<li>将分段脑电图数据导入脑电图软件 (文件 |加载现有数据集)。</li>
	<li>拒绝脑电图时代与振幅值超过80µV 在任何电极 (工具 |拒绝数据世纪 |拒绝数据 [所有方法])。</li>
	<li>保存预处理的脑电图数据 (文件 |将当前数据集另存为)。</li>
</ol>4. 脑电图微状态分析
注: 经典 k-均值聚类算法的一个改进版本用于微状态类分析16, 它包含一个自下而上的过程和一个自下而上的过程。在自底向上的过程中, 用空间相关作为聚类准则来识别群级微状态类。然后在自下而上的过程中, 每组每个主题的每个地形图都被分配到具有最大空间相关性的 EEG 微状态类。对于静止状态脑电图微状态分析, 地形图的极性通常被忽略。脑电图微状态类分析可以使用各种开源软件, 如 CARTOOL, sLORETA, 艾玛和 MapWin。下面提供的说明是特定的 EEGLAB 插件微状态。这个 EEGLAB 插件可以从 https://sccn.ucsd.edu/wiki/EEGLAB_Extensions_and_plug-ins 下载。
<ol><li>对于每个主题, 加载预处理的脑电图数据 (文件 |加载现有数据集), 将参考通道转换为普通平均引用 (工具 |重新参考), 和带通滤波器的脑电图数据之间的2和 20 Hz (工具 |筛选数据 |基本 FIR 过滤器 [新建, 默认])。</li>
	<li>确定每个主题中的四微状态映射 (工具 |微状态 |识别微状态地图)。在弹出对话框中, 为 "最小类数" 选择 "3", 为 "最大类数" 选择 6, 选择 "重新启动次数" 50, 选择 "要使用的最大映射数", 然后选择 "GFP 峰值" 和 "无极性" 选项。然后单击 "确定" 按钮。</li>
	<li>保存每个主题的脑电图数据后, 确定自己的微状态地图 (文件 |将当前数据集另存为)。</li>
	<li>导入最后一步中保存的所有主题的 EEG 数据集 (文件 |加载现有数据集)。</li>
	<li>识别组级微状态映射 (工具 |微状态 |平均微状态映射跨数据集)。在弹出对话框中, 选择 "选择平均值集" 选项中所有主题的数据集。在 "平均名称" 选项中, 给出组级微状态映射的名称。默认名称为 "GrandMean"。然后单击 "确定" 按钮。这将创建一个名为 "GrandMean" 的新数据集, 它存储组级微状态映射。</li>
	<li>根据他们的经典顺序手动排序四组级微状态地图 (剧情 |编辑微状态地图)。在弹出式中, 选择 "更多", 然后显示的地图数变为四。选择 "人类排序"。在弹出对话框中, 输入四组级微状态映射的新顺序。然后单击 "关闭"。</li>
	<li>排序每个主题的四微状态地图的顺序 (工具 |微状态 |按平均值对单个微状态映射进行排序)。</li>
	<li>保存每个主题的微状态参数 (工具 |微状态 |在数据集 [平均模板映射] 中量化微状态, 它将依次调用两个弹出对话框。<ol>
<li>在第一个对话框中, 选择所有主题的数据集。在第二个对话框中, 为选项 "类数" 选择 "4 类", 选择 "仅适用于 GFP 峰值" 和 "删除可能截断的微状态" 选项, 选择 "30" 作为 "标签平滑窗口 (ms)", 然后为 "非平滑性" 选择1。惩罚 "。然后单击 "确定"。存储微状态参数的 csv 文件将保存在计算机上。</li>
	</ol>
</li>
</ol>5. 欧米茄复杂性分析
<ol><li>使用自定义脚本保存每个纪元的 EEG 数据和每个主题的 ASCII 或 txt 格式。在补充材料中可以找到步骤5.1 的自定义脚本示例。 注: 如果计算了全局欧米茄复杂性, 则需要所有头皮电极的 EEG 数据以 ASCII 或 txt 格式导出。如果区域欧米茄复杂性被计算, 仅出口电极的 EEG 数据在那头皮区域。例如, 为了计算前欧米茄复杂度, 只导出前区电极的脑电图数据 (即 Fp1、Fp2、F7、F3、熔、F4 和 F8);为了计算后欧米茄复杂度, 只导出后区电极的脑电图数据 (即 T5、T6、P3、P4、Pz、O1 和 O2)。</li>
	<li>使用 sLORETA 软件17计算所有离散频率的欧米茄复杂度 (实用程序 |全局连接性)。此软件可在 http://www.uzh.ch/keyinst/loreta.htm 中使用。</li>
	<li>使用自定义脚本26计算每个频带的欧米茄复杂性。在我们的例子中, 计算以下八频带的欧米茄复杂度作为每个频率限制中的平均值, 这是三角洲 (0.5-3.5 hz), θ (4-7, 5 hz), alpha1 (8-10 hz), alpha2 (10.5-13.5 hz), beta1 (14-18 hz), beta2 (18.5-30 hz), gamma1 (30.5-48 hz), 和 gamma2 (52-80 Hz)4。在补充材料中可以找到步骤5.3 的自定义脚本示例。</li>
</ol>

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作者没有什么可透露的。

本文详细介绍了两种脑电图分析方法 (即微状态分析法和欧米茄复杂度法), 分别测量人脑的时间复杂度和空间复杂性。在议定书内有几个关键步骤应予提及。首先, 在计算微状态和欧米茄复杂度之前, 必须对 EEG 数据进行清理。其次, 在计算微状态和欧米茄复杂度之前, 应 remontaged 对脑电图数据进行平均参考。第三, 在微状态和欧米茄复杂度计算之前, 必须将连续脑电图数据分割成时代。每个纪元的长...

脑电图 (eeg) 已被广泛用于记录人脑的电活动在临床诊断和科学研究, 因为它是无创的, 低成本的, 有非常高的时间分辨率1。为了研究静止状态下的脑电图信号, 研究人员开发了许多脑电图技术 (如功率谱分析、功能连接分析)2、3。其中, 微状态分析和欧米茄复杂性分析可以很好地利用脑电信号4中固有的时空信息。
以往的研究表明, 尽管脑电信号的地形分布随着时间的推移在闭眼或睁开的静止状态下变化, 但序列的瞬时图显示了景观的不连续变化, 即稳定交替期在某些准稳定脑电图之间有短的过渡期地形5。微状态被定义作为这些情节与准稳定 EEG 地形, 为时在80和 120 ms1之间。由于不同的电势景观必须由不同的神经源产生, 这些微状态可能成为心理状态的基本块, 可以被认为是 "思想和情感的原子"6。采用现代模式分类算法, 连续观察到四个静止脑电图微状态类, 被标记为 a 类、B 类、C 类和 D 类7。此外, 研究人员发现, 这四微状态类静止脑电图数据与已知的功能系统密切联系在许多静止状态 fMRI (功能磁共振成像) 研究8,9.因此, 微状态分析为研究人脑的静止状态网络 (RSNs) 提供了一种新的方法。此外, 每微状态类的平均持续时间和发生频率, 四微状态地图的地形形状都受到一些脑部疾病4,10,11的影响,并且与流体智力12和个性13相关。
在另一方面, 多通道脑电图的传统功能连接只能描述两个头皮电极之间的功能连接, 从而无法评估整个头皮或某一脑区内的全球功能连通性。Wackermann (1996)14提出的欧米茄复杂性, 并通过结合主成分分析 (PCA) 和香农熵的方法计算, 已被用来量化宽带全球同步的空间分布的脑区。为了评估每个频带的欧米茄复杂度, 傅立叶变换通常是作为初始步骤25进行的。
微状态和欧米茄复杂性可以用来反映两个紧密相连的概念,即时态复杂性和空间复杂性4。由于微状态类代表人脑中的某些心理操作, 它们可以反映神经元振荡的时间结构。更低的持续时间和更高的发生率每秒必须表明更高的时态复杂性。欧米茄复杂性与大脑中独立神经源的数量呈正相关, 因此通常被认为是空间复杂度4的指标。
本文详细介绍了脑电图微状态分析和欧米茄复杂性分析的协议。脑电图微状态和欧米茄复杂性分析为测量脑活动的时间和空间复杂性提供了机会。

<table>
	<tbody>
		<tr>
			<td>ANT 20 channels EEG/ERP system</td>
			<td>ASA-Lab, ANT B.V., Netherlands</td>
			<td></td>
			<td>company web address: 
			http://www.ant-neuro.com/</td>
		</tr>
		<tr>
			<td>EEGLAB plugin for Microstates</td>
			<td>Thomas Koenig</td>
			<td></td>
			<td>https://sccn.ucsd.edu/wiki/EEGLAB_Extensions_and_plug-ins</td>
		</tr>
		<tr>
			<td>sLORETA</td>
			<td>Roberto D. Pascual-Marqui</td>
			<td></td>
			<td>http://www.uzh.ch/keyinst/loreta.htm</td>
		</tr>
		<tr>
			<td>MATLAB 2010a</td>
			<td>The MathWorks Inc.</td>
			<td></td>
			<td>company web address: 
			http://www.mathworks.com/</td>
		</tr>
		<tr>
			<td>eeglab</td>
			<td>Swartz Center for Computational Neuroscience, University of California, San Diego</td>
			<td></td>
			<td>https://sccn.ucsd.edu/eeglab/index.php</td>
		</tr>
	</tbody>
</table>

microstate and omega complexity analyses of the resting-state electroencephalography

微状态和欧米茄的复杂性是两个无参考脑电图 (eeg) 措施, 可以代表的时间和空间复杂的脑电图数据, 并已广泛用于调查神经机制的一些脑部疾病。本文的目的是描述脑电图微状态和欧米茄复杂性分析的基本协议。这两种方法的主要优点是可以消除传统频谱分析固有的参考依赖问题。此外, 微状态分析充分利用了静止状态脑电图的高时间分辨力, 得到的四微状态类可以分别匹配相应的休眠状态网络。欧米茄复杂性的特点是整个大脑或特定大脑区域的空间复杂度, 这与传统的复杂度测量方法相比, 在单个信道中的信号复杂度具有明显的优越性。这两种脑电图测量方法可以相互补充, 分别从时间和空间领域对大脑的复杂性进行研究。

脑电图微状态
图 1显示了大平均规范化的微状态映射。这里所确定的四微状态类的电势景观与以前的研究4中所发现的类似。
表 1显示了健康受试者微状态参数的平均值和标准差 (SD)。对于微状态 a 类, 其发生率为 3.44, 1.29 次/秒, 持?...

Watch this Scientific Journal Video about Microstate and Omega Complexity Analyses of the Resting-state Electroencephalography at JoVE.com

Microstate and Omega Complexity Analyses of the Resting-state Electroencephalography

本文描述了基于脑电图 (eeg) 微状态分析和欧米茄复杂性分析的协议, 这是两种无参考脑电图的方法, 对探讨脑功能紊乱的神经机制具有十分重要的价值。

静止状态脑电图的微状态和欧米茄复杂性分析

Microstate and omega complexity are two reference-free electroencephalography (EEG) measures that can represent the temporal and spatial complexities of ...

microstate-omega-complexity-analyses-resting-state

Research

JoVE Journal

Neuroscience

10.1K 次观看.  Peking University People's Hospital. 本文描述了基于脑电图 (eeg) 微状态分析和欧米茄复杂性分析的协议, 这是两种无参考脑电图的方法, 对探讨脑功能紊乱的神经机制具有十分重要的价值。

视频: Microstate and Omega Complexity Analyses of the Resting-state Electroencephalography

Investigating Social Cognition in Infants and Adults Using Dense Array Electroencephalography (dEEG)

Dense array electroencephalography (dEEG), which provides a non-invasive window for measuring brain activity and a temporal resolution unsurpassed by any other current brain imaging technology1,2, is being used increasingly in the study of social cognitive functioning in infants and adults. While dEEG is enabling researchers to examine brain activity patterns with unprecedented levels of sensitivity, conventional EEG recording systems continue to face certain limitations, including 1) poor spatial resolution and source localization3,4,2) the physical discomfort for test subjects of enduring the individual application of numerous electrodes to the surface of the scalp, and 3) the complexity for researchers of learning to use multiple software packages to collect and process data. Here we present an overview of an established methodology that represents a significant improvement on conventional methodologies for studying EEG in infants and adults. Although several analytical software techniques can be used to establish indirect indices of source localization to improve the spatial resolution of dEEG, the HydroCel Geodesic Sensor Net (HCGSN) by Electrical Geodesics, Inc. (EGI), a dense sensory array that maintains equal distances among adjacent recording electrodes on all surfaces of the scalp, further enhances spatial resolution4,5,6 compared to standard dEEG systems. The sponge-based HCGSN can be applied rapidly and without scalp abrasion, making it ideal for use with adults7,8, children9,10,11, and infants12, in both research and clinical4,5,6,13,14,15 settings. This feature allows for considerable cost and time savings by decreasing the average net application time compared to other dEEG systems. Moreover, the HCGSN includes unified, seamless software applications for all phases of data, greatly simplifying the collection, processing, and analysis of dEEG data.
The HCGSN features a low-profile electrode pedestal, which, when filled with electrolyte solution, creates a sealed microenvironment and an electrode-scalp interface. In all Geodesic dEEG systems, EEG sensors detect changes in voltage originating from the participant's scalp, along with a small amount of electrical noise originating from the room environment. Electrical signals from all sensors of the Geodesic sensor net are received simultaneously by the amplifier, where they are automatically processed, packaged, and sent to the data-acquisition computer (DAC). Once received by the DAC, scalp electrical activity can be isolated from artifacts for analysis using the filtering and artifact detection tools included in the EGI software. Typically, the HCGSN can be used continuously for only up to two hours because the electrolyte solution dries out over time, gradually decreasing the quality of the scalp-electrode interface.
In the Parent-Infant Research Lab at the University of Toronto, we are using dEEG to study social cognitive processes including memory, emotion, goals, intentionality, anticipation, and executive functioning in both adult and infant participants.

Investigating Social Cognition in Infants and Adults Using Dense Array Electroencephalography dEEG

Dense array electroencephalography is being used increasingly to study social cognitive functions in infants and adults. Here we present an established methodology that represents a significant improvement on conventional methodologies for studying EEG in infants and adults.

调查婴儿和成人使用密集阵脑电图的社会认知（ D脑电图）

Dense array electroencephalography (dEEG), which provides a non-invasive window for measuring brain activity and a temporal resolution unsurpassed by any ...

科研

神经科学

Morphometric Analyses of Retinal Sections

Morphometric analyses of retinal sections have been used in examining retinal diseases. For examples, neuronal cells were significantly lost in the retinal ganglion cell layer (RGCL) in rat models with N-methyl-D-aspartate (NMDA)&#x2013;induced excitotoxicity1, retinal ischemia-reperfusion injury2 and glaucoma3. Reduction of INL and inner plexiform layer (IPL) thicknesses were reversed with citicoline treatment in rats' eyes subjected to kainic acid-mediated glutamate excitotoxicity4. Alteration of RGC density and soma sizes were observed with different drug treatments in eyes with elevated intraocular pressure3,5,6. Therefore, having objective methods of analyzing the retinal morphometries may be of great significance in evaluating retinal pathologies and the effectiveness of therapeutic strategies.
The retinal structure is multi-layers and several different kinds of neurons exist in the retina. The morphometric parameters of retina such as cell number, cell size and thickness of different layers are more complex than the cell culture system. Early on, these parameters can be detected using other commercial imaging software. The values are normally of relative value, and changing to the precise value may need further accurate calculation. Also, the tracing of the cell size and morphology may not be accurate and sensitive enough for statistic analysis, especially in the chronic glaucoma model. The measurements used in this protocol provided a more precise and easy way. And the absolute length of the line and size of the cell can be reported directly and easy to be copied to other files. For example, we traced the margin of the inner and outer most nuclei in the INL and formed a line then using the software to draw a 90 degree angle to measure the thickness. While without the help of the software, the line maybe oblique and the changing of retinal thickness may not be repeatable among individual observers. In addition, the number and density of RGCs can also be quantified. This protocol successfully decreases the variability in quantitating features of the retina, increases the sensitivity in detecting minimal changes.
 	This video will demonstrate three types of morphometric analyses of the retinal sections. They include measuring the INL thickness, quantifying the number of RGCs and measuring the sizes of RGCs in absolute value. These three analyses are carried out with Stereo Investigator (MBF Bioscience &mdash; MicroBrightField, Inc.). The technique can offer a simple but scientific platform for morphometric analyses.

This video demonstrates three types of morphometric analyses of the retina, which include measuring the inner nuclear layer thickness, quantifying the number of retinal ganglion cells (RGCs) and measuring the sizes of RGCs. The technique can offer a simple but scientific platform for morphometric analyses.

视网膜节的形态分析

Morphometric analyses of retinal sections have been used in examining retinal diseases. For examples, neuronal cells were significantly lost in the ...

Optogenetic Perturbation of Neural Activity with Laser Illumination in Semi-intact Drosophila Larvae in Motion

Drosophila larval locomotion is a splendid model system in developmental and physiological neuroscience, by virtue of the genetic accessibility of the underlying neuronal components in the circuits1-6. Application of optogenetics7,8 in the larval neural circuit allows us to manipulate neuronal activity in spatially and temporally patterned ways9-13. Typically, specimens are broadly illuminated with a mercury lamp or LED, so specificity of the target neurons is controlled by binary gene expression systems such as the Gal4-UAS system14,15. In this work, to improve the spatial resolution to "sub-genetic resolution", we locally illuminated a subset of neurons in the ventral nerve cord using lasers implemented in a conventional confocal microscope. While monitoring the motion of the body wall of the semi-intact larvae, we interactively activated or inhibited neural activity with channelrhodopsin16,17 or halorhodopsin18-20, respectively. By spatially and temporally restricted illumination of the neural tissue, we can manipulate the activity of specific neurons in the circuit at a specific phase of behavior. This method is useful for studying the relationship between the activities of a local neural assembly in the ventral nerve cord and the spatiotemporal pattern of motor output.

Optogenetic Perturbation of Neural Activity with Laser Illumination in Semi-intact Drosophila Larvae in Motion

Here we describe a protocol for optogenetic manipulation of motoneuronal activity while monitoring changes in motor output (muscle contraction) in semi-intact Drosophila larvae using lasers within a conventional confocal microscope. This technique enables researchers to achieve local perturbation of neural activity in a few neuromeres to elucidate the dynamics of motor circuits.

光遗传学的扰动激光照射半完好的神经活动<em&gt;果蝇</em&gt;幼虫运动

Drosophila larval locomotion is a splendid model system in developmental and physiological neuroscience, by virtue of the genetic accessibility of the ...

Investigating the Function of Deep Cortical and Subcortical Structures Using Stereotactic Electroencephalography: Lessons from the Anterior Cingulate Cortex

Stereotactic Electroencephalography (SEEG) is a technique used to localize seizure foci in patients with medically intractable epilepsy. This procedure involves the chronic placement of multiple depth electrodes into regions of the brain typically inaccessible via subdural grid electrode placement. SEEG thus provides a unique opportunity to investigate brain function. In this paper we demonstrate how SEEG can be used to investigate the role of the dorsal anterior cingulate cortex (dACC) in cognitive control. We include a description of the SEEG procedure, demonstrating the surgical placement of the electrodes. We describe the components and process required to record local field potential (LFP) data from consenting subjects while they are engaged in a behavioral task. In the example provided, subjects play a cognitive interference task, and we demonstrate how signals are recorded and analyzed from electrodes in the dorsal anterior cingulate cortex, an area intimately involved in decision-making. We conclude with further suggestions of ways in which this method can be used for investigating human cognitive processes.

Stereotactic Electroencephalography (SEEG) is an operative technique used in epilepsy surgery to help localize seizure foci. It also affords a unique opportunity to investigate brain function. Here we describe how SEEG can be used to investigate cognitive processes in human subjects.

调查深皮层的功​​能和皮层下结构采用立体定向脑电图:由前扣带皮层的经验教训

Stereotactic Electroencephalography (SEEG) is a technique used to localize seizure foci in patients with medically intractable epilepsy. This procedure ...

Assessment of Hippocampal Dendritic Complexity in Aged Mice Using the Golgi-Cox Method

Dendritic spines are the protuberances from the neuronal dendritic shafts that contain&#160; excitatory synapses. The morphological and branching variations of the neuronal dendrites within the hippocampus are implicated in cognition and memory formation. There are several approaches to Golgi staining, all of which have been useful for determining the morphological characteristics of dendritic arbors and produce a clear background. The present Golgi-Cox method, (a slight variation of the protocol that is provided with a commercial Golgi staining kit), was designed to assess how a relatively low dose of the chemotherapeutic drug 5-flurouracil (5-Fu) would affect dendritic morphology, the number of spines, and the complexity of arborization within the hippocampus. The 5-Fu significantly modulated the dendritic complexity and decreased the spine density throughout the hippocampus in a region-specific manner. The data presented show that the Golgi staining method effectively stained the mature neurons in the CA1, the CA3, and the dentate gyrus (DG) of the hippocampus. This protocol reports the details for each step so that other researchers can reliably stain tissue throughout the brain with high quality results and minimal troubleshooting.

Here we present a Golgi-Cox protocol in extensive detail. This reliable tissue stain method allows for a high-quality assessment of the cytoarchitecture in the hippocampus, and throughout the entire brain, with minimal troubleshooting.

使用高尔基Cox方法评估老年小鼠海马树突状复杂性

Dendritic spines are the protuberances from the neuronal dendritic shafts that contain&#160; excitatory synapses. The morphological and branching ...

Concurrent Recording of Co-localized Electroencephalography and Local Field Potential in Rodent

Although electroencephalography (EEG) is widely used as a non-invasive technique for recording neural activities of the brain, our understanding of the neurogenesis of EEG is still very limited. Local field potentials (LFPs) recorded via a multi-laminar microelectrode can provide a more detailed account of simultaneous neural activity across different cortical layers in the neocortex, but the technique is invasive. Combining EEG and LFP measurements in a pre-clinical model can greatly enhance understanding of the neural mechanisms involved in the generation of EEG signals, and facilitate the derivation of a more realistic and biologically accurate mathematical model of EEG. A simple procedure for acquiring concurrent and co-localized EEG and multi-laminar LFP signals in the anesthetized rodent is presented here. We also investigated whether EEG signals were significantly affected by a burr hole drilled in the skull for the insertion of a microelectrode. Our results suggest that the burr hole has a negligible impact on EEG recordings.

This protocol describes a simple method for concurrent recording of co-localized electroencephalography (EEG) and multi-laminar local field potential in an anesthetized rat. A burr hole drilled in the skull for the insertion of a microelectrode is shown to produce negligible distortion of the EEG signal.

啮齿动物联合局部脑电图和局部电位的同步记录

Although electroencephalography (EEG) is widely used as a non-invasive technique for recording neural activities of the brain, our understanding of the ...

Quantitative Analysis of Neuronal Dendritic Arborization Complexity in Drosophila

Dendrites are the branched projections of a neuron, and dendritic morphology reflects synaptic organization during the development of the nervous system. Drosophila larval neuronal dendritic arborization (da) is an ideal model for studying morphogenesis of neural dendrites and gene function in the development of nervous system. There are four classes of da neurons. Class IV is the most complex with a branching pattern that covers almost the entire area of the larval body wall. We have previously characterized the effect of silencing the Drosophila ortholog of SOX5 on class IV neuronal dendritic arborization complexity (NDAC) using four parameters: the length of dendrites, the surface area of dendrite coverage, the total number of branches, and the branching structure. This protocol presents the workflow of NDAC quantitative analysis, consisting of larval dissection, confocal microscopy, and image analysis procedures using ImageJ software. Further insight into da neuronal development and its underlying mechanisms will improve the understanding of neuronal function and provide clues about the fundamental causes of neurological and neurodevelopmental disorders.

Quantitative Analysis of Neuronal Dendritic Arborization Complexity in Drosophila

This protocol focuses on quantitative analysis of neuronal dendritic arborization complexity (NDAC) in Drosophila, which can be used for studies of dendritic morphogenesis.

嗜德维人神经元树突状树突状复杂度的定量分析

Dendrites are the branched projections of a neuron, and dendritic morphology reflects synaptic organization during the development of the nervous system. ...

Combined Transcranial Magnetic Stimulation and Electroencephalography of the Dorsolateral Prefrontal Cortex

Transcranial magnetic stimulation (TMS) is a non-invasive method that produces neural excitation in the cortex by means of brief, time-varying magnetic field pulses. The initiation of cortical activation or its modulation depends on the background activation of the neurons of the cortical region activated, the characteristics of the coil, its position and its orientation with respect to the head. TMS combined with simultaneous electrocephalography (EEG) and neuronavigation (nTMS-EEG) allows for the assessment of cortico-cortical excitability and connectivity in almost all cortical areas in a reproducible manner. This advance makes nTMS-EEG a powerful tool that can accurately assess brain dynamics and neurophysiology in test-retest paradigms that are required for clinical trials. Limitations of this method include artifacts that cover the initial brain reactivity to stimulation. Thus, the process of removing artifacts may also extract valuable information. Moreover, the optimal parameters for dorsolateral prefrontal (DLPFC) stimulation are not fully known and current protocols utilize variations from the motor cortex (M1) stimulation paradigms. However, evolving nTMS-EEG designs hope to address these issues. The protocol presented here introduces some standard practices for assessing neurophysiological functioning from stimulation to the DLPFC that can be applied in patients with treatment resistant psychiatric disorders that receive treatment such as transcranial direct current stimulation (tDCS), repetitive transcranial magnetic stimulation (rTMS), magnetic seizure therapy (MST) or electroconvulsive therapy (ECT).

The protocol presented here is for TMS-EEG studies utilizing intracortical excitability test-retest design paradigms. The intent of the protocol is to produce reliable and reproducible cortical excitability measures for assessing neurophysiological functioning related to therapeutic interventions in the treatment of neuropsychiatric diseases such as major depression.

侧前额叶皮质联合经颅磁刺激与脑电图的研究

Transcranial magnetic stimulation (TMS) is a non-invasive method that produces neural excitation in the cortex by means of brief, time-varying magnetic ...

Collection of Frozen Rodent Brain Regions for Downstream Analyses

As our understanding of neurobiology has progressed, molecular analyses are often performed on small brain areas such as the medial prefrontal cortex (mPFC) or nucleus accumbens. The challenge in this work is to dissect the correct area while preserving the microenvironment to be examined. In this paper, we describe a simple, low-cost method using resources readily available in most labs. This method preserves nucleic acid and proteins by keeping the tissue frozen throughout the process. Brains are cut into 0.5&#8211;1.0 mm sections using a brain matrix and arranged on a frozen glass plate. Landmarks within each section are compared to a reference, such as the Allen Mouse Brain Atlas, and regions are dissected using a cold scalpel or biopsy punch. Tissue is then stored at -80 &#176;C until use. Through this process rat and mouse mPFC, nucleus accumbens, dorsal and ventral hippocampus and other regions have been successfully analyzed using qRT-PCR and Western assays. This method is limited to brain regions that can be identified by clear landmarks.

This procedure describes the collection of discrete frozen brain regions to obtain high-quality protein and RNA using inexpensive and commonly available tools.

用于下游分析的冷冻啮齿动物大脑区域集合

As our understanding of neurobiology has progressed, molecular analyses are often performed on small brain areas such as the medial prefrontal cortex ...

Preparation of the Rat Vocal Fold for Neuromuscular Analyses

The purpose of this tutorial is to describe the preparation of the rat vocal fold for histochemical neuromuscular study. This protocol outlines procedures for rat laryngeal dissection, flash-freezing, and cryosectioning of the vocal folds. This study describes how to cryosection vocal folds in both longitudinal and cross-sectional planes. A novelty of this protocol is the laryngeal tracking during cryosectioning that ensures accurate identification of the intrinsic laryngeal muscles and reduces the chance of tissue loss. Figures demonstrate the progressive cryosectioning in both planes. Twenty-nine rat hemi-larynges were cryosectioned and tracked from the emergence of the thyroid cartilage to the appearance of the first section that included the full vocal fold. The full vocal fold was visualized for all animals in both planes. There was high variability in the distance from the appearance of the thyroid cartilage to the appearance of the full vocal fold in both planes. Weight was not correlated to depth of laryngeal landmarks, suggesting individual variability and other factors related to tissue preparation may be responsible for the high variability in the appearance of landmarks during sectioning. This study details a methodology and presents morphological data for preparing the rat vocal fold for histochemical neuromuscular investigation. Due to high individual variability, laryngeal landmarks should be closely tracked during cryosectioning to prevent oversectioning tissue and tissue loss. The use of a consistent methodology, including adequate tissue preparation and awareness of landmarks within the rat larynx, will assist with consistent results across studies and aid new researchers interested in using the rat vocal fold as a model to investigate laryngeal neuromuscular mechanisms.

This protocol describes methods used to prepare rat vocal folds for histochemical neuromuscular study.

用于神经肌肉分析的大鼠声带的制备

The purpose of this tutorial is to describe the preparation of the rat vocal fold for histochemical neuromuscular study. This protocol outlines procedures ...