Name: using neuron spiking activity to trigger closed-loop stimuli in neurophysiological experiments
Uploaded: 2019-11-12T11:00:00
Description: Watch this Scientific Journal Video about Using Neuron Spiking Activity to Trigger Closed-Loop Stimuli in Neurophysiological Experiments at JoVE.com

这项工作得到了NSERC发现公司对AL和AG的资助。

此处描述的所有程序均根据莱特布里奇大学动物福利委员会批准的动物研究协议进行。
1. 外科
注：用于植入神经生理学记录的探针的手术程序已在其他出版物24、25、26中发表。闭环刺激手术的确切细节取决于所使用的记录探头类型和目标的大脑区域。然而，在大多数情况下，典型的手?...

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此处描述的协议演示如何使用标准神经生理记录系统执行闭环刺激。该协议允许在计算机科学方面拥有有限专业知识的神经科学家以很少的成本快速实施各种闭环实验。这种实验经常是必要的，以研究在大脑的因果关系。
在准备动物并安装软件（步骤 1 和 2）后，闭环实验由两个单独的阶段组成。首先，初始数据采集（步骤3）收集数据，以定义与单个神经元活动对应的模板?...

该协议的目的是演示如何在神经生理学实验中实现闭环刺激。神经科学中闭环实验的典型设置涉及基于神经元活动的在线读出触发刺激。这反过来又导致大脑活动的变化，从而关闭反馈循环1，2。这种闭环实验比标准开环设置具有多种优势，特别是与光遗传学结合时，这允许研究人员针对特定的神经元子集。例如，Siegle和Wilson使用闭环操作来研究ta振荡在信息处理中的作用3。他们证明，刺激海马神经元在塔振荡的下降阶段对行为的影响与在上升阶段应用相同的刺激不同。闭环实验在临床前研究也越来越重要。例如，多种癫痫研究表明，癫痫发作时触发的神经元刺激是降低癫痫发作严重程度的一种有效方法。此外，自动癫痫检测系统和或送治疗7、8的系统对癫痫患者9、10、11、12显示出显著益处。闭环方法迅速发展的另一个应用领域是使用皮质脑机接口控制神经假体。这是因为向假肢装置的用户提供即时反馈可显著提高准确性和能力13。
近年来，一些实验室已经开发出定制系统，用于同步电子记录神经元活动和在闭环系统14，15，16，17，18的刺激。尽管其中许多设置具有令人印象深刻的特性，但在其他实验室中实现它们并不总是那么容易。这是因为系统通常需要经验丰富的技术人员来组装所需的电子元件和其他必要的硬件和软件组件。
因此，为了便于在神经科学研究中采用闭环实验，本文提供了一个协议和Matlab代码，将开环电生理记录设置19、20、21、22转换为闭环系统2、6、23。该协议旨在与数字 Lynx 记录硬件配合使用，这是一种用于神经元群记录的常用实验室系统。典型的实验包括：1） 记录 5-20 分钟的尖峰数据;2） 尖峰排序创建神经元模板;3） 使用这些模板对神经活动模式进行在线检测;4） 检测到用户指定的模式时触发刺激或实验事件。

<table>
	<tbody>
		<tr>
			<td>Baytril</td>
			<td>Bayer, Mississauga, CA</td>
			<td>DIN 02169428</td>
			<td>antibiotic; 50 mg/mL</td>
		</tr>
		<tr>
			<td>Cheetah 6.4</td>
			<td>NeuraLynx, Tucson, AZ</td>
			<td>6.4.0.beta</td>
			<td>Software interfaces for data acquisition&#160;</td>
		</tr>
		<tr>
			<td>Digital Lynx 4SX</td>
			<td>NeuraLynx, Tucson, AZ</td>
			<td>4SX</td>
			<td>recording equipment</td>
		</tr>
		<tr>
			<td>Headstage transmitter</td>
			<td>TBSI</td>
			<td>B10-3163-GK</td>
			<td>transmits the neural signal to the receiver</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Fresenius Kabi, Toronto, CA</td>
			<td>DIN 02237518</td>
			<td>inhalation anesthetic</td>
		</tr>
		<tr>
			<td>Jet Denture Powder &#38; Liqud</td>
			<td>Lang Dental, Wheeling, US</td>
			<td>1230</td>
			<td>dental acrylic</td>
		</tr>
		<tr>
			<td>Lacri-Lube</td>
			<td>Allergan, Markham, CA</td>
			<td>DIN 00210889</td>
			<td>eye ointment</td>
		</tr>
		<tr>
			<td>Lido-2</td>
			<td>Rafter 8, Calgary</td>
			<td>DIN 00654639</td>
			<td>local anesthetic; 20 mg/mL</td>
		</tr>
		<tr>
			<td>Matlab</td>
			<td>Mathworks</td>
			<td>R2018b</td>
			<td>software for signal processing and triggering external events</td>
		</tr>
		<tr>
			<td>Metacam</td>
			<td>Boehringer, Ingelheim, DE</td>
			<td>DIN 02240463</td>
			<td>analgesic; 5 mg/mL</td>
		</tr>
		<tr>
			<td>Netcom</td>
			<td>NeuraLynx</td>
			<td>v1</td>
			<td>Application Programming Interface (API) that communicates with Cheetah</td>
		</tr>
		<tr>
			<td>Silicone probe</td>
			<td>Cambridge Neurotech</td>
			<td>ASSY-156-DBC2</td>
			<td>implanted device</td>
		</tr>
		<tr>
			<td>SpikeSort 3D&#160;</td>
			<td>NeuraLynx, Tucson, AZ</td>
			<td>SS3D</td>
			<td>spike waveform-to-cell classification tools</td>
		</tr>
		<tr>
			<td>Wireless Radio Receiver</td>
			<td>TBSI</td>
			<td>911-1062-00</td>
			<td>transmits the neural signal to the Digital Lynx</td>
		</tr>
	</tbody>
</table>

using neuron spiking activity to trigger closed-loop stimuli in neurophysiological experiments

闭环神经生理学系统使用神经元活动模式来触发刺激，进而影响大脑活动。这种闭环系统已经在临床应用中找到，是基础脑研究的重要工具。最近一个特别有趣的发展是闭环方法与光遗传学的整合，这样神经元活动的特定模式可以触发选定神经元群的光学刺激。然而，为闭环实验建立电生理系统是很困难的。在这里，提供了一个随时应用的Matlab代码，用于根据单神经元或多个神经元的活动触发刺激。此示例代码可根据个人需求轻松修改。例如，它显示了如何触发声音刺激以及如何更改声音以触发连接到 PC 串行端口的外部设备。提出的协议设计用于动物研究（Neuralynx）的流行神经元记录系统。闭环刺激的实现在醒着的大鼠中得到证明。

Fisher-Brown挪威大鼠在现场出生和长大，在实验前两周习惯处理。一个记录驱动器被手术植入，类似于前面描述的方法28，29，30，31，32，33，34。使用数字采集系统在32 kHz下记录神经元信号。神经元信号首先被放大，使用统?...

Watch this Scientific Journal Video about Using Neuron Spiking Activity to Trigger Closed-Loop Stimuli in Neurophysiological Experiments at JoVE.com

Using Neuron Spiking Activity to Trigger Closed-Loop Stimuli in Neurophysiological Experiments

该协议演示如何使用电生理系统进行由神经元活动模式触发的闭环刺激。还提供可轻松针对不同刺激设备修改的示例 Matlab 代码。

在神经生理学实验中利用神经元刺激活性触发闭环刺激

Closed-loop neurophysiological systems use patterns of neuronal activity to trigger stimuli, which in turn affect brain activity. Such closed-loop systems ...

这个标准的神经生理记录系统可用于进行闭环刺激，这对于研究大脑中的因果相互作用非常有用。该协议允许在计算机科学领域专业知识有限的神经科学家以很少的成本快速实施各种闭环实验。闭环实验对基础科学和研究越来越重要。例如，多发癫痫研究表明，在癫痫发作时神经元刺激触发是降低癫痫发作严重程度的有效方法。为了便于在神经科学研究中采用闭环实验，该协议提供了MATLAB代码，用于将闭环电生理记录系统转换为闭环系统。这种闭环刺激的实现在清醒的老鼠中表现出来。要安装猎豹 6.4 数据采集系统，请打开包含与猎豹采集系统交互的库的软件网页，然后安装 SpikeSort-3D。安装网通开发包。登录到 MATLAB 帐户以选择正确的许可证、版本和操作系统。然后，下载联机事件触发的库，然后将文件提取到文档 MATLAB 文件夹。对于数据采集，请打开 Cheetah 软件并记录尖峰数据以填充模板波形。几分钟后，停止数据采集，对记录的数据执行 SpikeSort。接下来，打开 SpikeSort3-D 并单击文件菜单并加载峰值文件从包含记录数据的文件夹中选择峰值文件。然后，单击群集菜单，使用 KlustaKwik 自动群集，然后运行。要执行闭环实验，请恢复猎豹中的数据采集，然后打开 MATLAB。打开闭合循环。M 并单击"运行"。或者，在命令窗口中执行闭环。浏览到录制文件夹并选择其中一个尖峰数据文件，以加载初始录制中定义的尖峰信息。单击绘制的波形下相应的复选框，选择一个或多个将触发刺激的神经元。定义通过输入 min 匹配文本框中的整数来触发刺激的最小神经元数，并定义通过在窗口文本框中输入数字来将匹配不同波形的峰值视为共动的时间窗口。然后，单击"发送"以根据所选神经元的尖峰活动开始事件的在线触发。在这个具有代表性的实验中，老鼠被放在一个花盆上，花盆在行为实验中休息时是一个熟悉的休息场所。在这里，可以观察到记录软件和MATLAB程序的代表性同时运行，该程序显示实时获取的峰值波形。当检测到预定义触发群集的峰值时，在 MATLAB 图形窗口中用粗体虚线显示波形，从而触发音调并提供闭环系统。在图中，显示了一个代表性直方图，说明生成人工尖峰的时间与触发触发信号的时间之间的延迟。在准备动物并安装软件后，闭环实验包括两个主要阶段：初始数据采集和闭环刺激，在此期间尖峰触发外部刺激。该协议还允许基于多个神经元的活动来治疗刺激，这对于解决大脑中的信息处理问题非常重要。此 MATLAB 解决方案的优点是，它具有低成本和灵活性，可以定义触发刺激的活动模式，从而有助于研究各种神经科学问题。

Research

JoVE Journal

Neuroscience

An Experimental Platform to Study the Closed-loop Performance of Brain-machine Interfaces

研究脑机接口的闭环性能的实验平台

The non-stationary nature and variability of neuronal signals is a fundamental problem in brain-machine interfacing. We developed a brain-machine ...

科研

神经科学

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

多单位记录方法来描述蝗虫的神经活动（<em&gt; Schistocerca美洲</em&gt;）嗅觉电路

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

The Neuromuscular Junction: Measuring Synapse Size, Fragmentation and Changes in Synaptic Protein Density Using Confocal Fluorescence Microscopy

神经肌肉接头处：测量突触大小，碎片和改变突触蛋白密度利用共聚焦荧光显微镜

The neuromuscular junction (NMJ) is the large, cholinergic relay synapse through which mammalian motor neurons control voluntary muscle contraction. ...

Real-time Electrophysiology: Using Closed-loop Protocols to Probe Neuronal Dynamics and Beyond

实时电：采用闭环协议来探测神经动力学和超越

Experimental neuroscience is witnessing an increased interest in the development and application of novel and often complex, closed-loop protocols, where ...

Closed-loop Neuro-robotic Experiments to Test Computational Properties of Neuronal Networks

闭环神经机器人实验来检验神经网络的计算性能

Information coding in the Central Nervous System (CNS) remains unexplored. There is mounting evidence that, even at a very low level, the representation ...

Immunohistochemical Visualization of Hippocampal Neuron Activity After Spatial Learning in a Mouse Model of Neurodevelopmental Disorders

海马神经元活动的可视化免疫组化空间后学习神经发育障碍的小鼠模型

Induction of phosphorylated extracellular-regulated kinase (pERK) is a reliable molecular readout of learning-dependent neuronal activation. Here, we ...

Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals

使用单一感器录音来检测臭虫的嗅觉神经元的反应，以信息化学物质

The insect olfactory system plays an important role in detecting semiochemicals in the environment. In particular, the antennal sensilla which house ...

Neuron-Macrophage Co-cultures to Activate Macrophages Secreting Molecular Factors with Neurite Outgrowth Activity

神经元-巨噬细胞共培养活化巨噬细胞分泌突起活性的分子因子

There is strong evidence that macrophages can participate in the regeneration or repair of injured nervous system. Here, we describe a protocol in which ...

Semi-Quantitative Determination of Dopaminergic Neuron Density in the Substantia Nigra of Rodent Models using Automated Image Analysis

使用自动图像分析对啮齿动物模型的亚斯坦蒂娅尼格拉多巴明神经元密度进行半定量测定

Estimation of the number of dopaminergic neurons in the substantia nigra is a key method in pre-clinical Parkinson's disease research. Currently, unbiased ...

In vivo Calcium Imaging of Mouse Geniculate Ganglion Neuron Responses to Taste Stimuli

在活体钙成像小鼠致病性甘里昂神经元对味觉刺激的反应

Within the last ten years, advances in genetically encoded calcium indicators (GECIs) have promoted a revolution in in vivo functional imaging. Using ...