Name: modeling fast-scan cyclic voltammetry data from electrically stimulated dopamine neurotransmission data using qnsim1.0
Uploaded: 2017-06-05T12:30:00.000+00:00
Duration: 7 min 41 s
Description: Watch this Scientific Journal Video about Modeling Fast-scan Cyclic Voltammetry Data from Electrically Stimulated Dopamine Neurotransmission Data Using QNsim1.0 at JoVE.com

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1.安装，数据准备和启动QNsim1.0 <ol><li>下载"QNsim1.0.zip"（作为补充提供），并将其解压缩到所需的目录。 </li><li>通过组织电子表格准备刺激的DA响应数据，用于通过软件程序进行建模，其中每列包含转换为μM DA浓度的时间DA响应。将此（.xlsx）文件另存为与程序文件相同的目录。 注意:电子表格可以包含单个实验中的多个响应或包含固定数量响应的多个研究（有关示例，请参阅"Sample.xlsx"）。 </li><li>打开编程环境软件，导航到"当前文件夹"窗口中的"QNSim1.0"目录，打开名为"Initialization.m。"的文件。单击"运行"以启动初始化屏幕（ 图1 ）。 </li></ol> 图1 :初始化屏幕。该屏幕允许用户通过输入以前的* .mat项目文件（左）继续现有的项目，或者从受保护的DA神经传递（右）的存储数据开始一个新的项目。提供文本框以输入包含数据的* .xlsx文件名，数据的几个基本描述符，以及* .mat项目文件名，其中包含与项目相关的所有数据。 <a href="http://ecsource.jove.com/files/ftp_upload/55595/55595fig1large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> 2.仿真环境的初始化注意:初始化屏幕允许用户开始一个新项目（步骤2.1）或继续以前保存的项目（步骤2.2）。 <ol><li> 选项1:开始一个新项目<ol><li>在"新建项目"部分下，将包含项目的DA响应数据（ 例如 Sample.xlsx）的电子表格文件的名称输入到[DA]数据文件文本框中。 注意:文件必须与程序文件在同一个文件夹中。 </li><li>在模拟时间旁边，输入相对于刺激开始的数据收集的开始和结束（以秒为单位）的时间点。例如，如果数据采集在刺激前5秒开始，并在35秒后结束，如Sample.xlsx中输入"-5"和"35"。 </li><li>在每个研究中输入相应文本框旁边的答复数。 </li><li>在采样间隔旁边，输入实验数据采样间隔（以秒为单位）。 </li><li>另存为（.mat）旁边，指定文件名，包括要保存项目的文件类型（.mat）。 注意:这是要访问的文件来创建模拟，所有工作都可以保存到此项目文件中。 </li><li>单击创建新项目以启动模拟器窗口（ 图2 ）。 </li></ol></li><li> 选项2:继续以前的项目 <ol><li>在"初始化"窗口的"继续上一个项目"部分中，输入先前开始的项目的.mat文件名（ 例如， Sample.mat）。 注意:该文件必须包含在与软件程序文件相同的文件夹中。 </li><li>单击加载现有项目启动模拟器窗口（ 图2 ）。 </li></ol></li></ol><img alt="图2" class="xfigimg" src="/files/ftp_upload/55595/55595fig2.jpg" /> 图2 :模拟器屏幕。模拟器屏幕允许用户在D中选择实验数据，并在E中调整模拟参数以对实验进行建模数据。 （ A，B和C ）的视觉检查可以帮助模拟参数的细化，以使模拟（蓝色虚线）模拟实验数据（粗绿色线）。这里，（ A ）包含实验和模拟DA浓度对时间数据。 （ B ）是有助于估计刺激后DA释放的图表。 （ C ）是DA浓度对来自A的时间数据或诱发溢出率（EO）的实验和模拟的一阶导数的曲线图。理论上，EO的比率是释放再摄取速率的平衡，其叠加在该图上。 <a href="http://ecsource.jove.com/files/ftp_upload/55595/55595fig2large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> 3.使用模拟器窗口注意:模拟器屏幕允许选择实验数据到模型（步骤3.1），调整参数以模拟实验DA响应数据（步骤3.2），为每个DA响应保存/加载建模参数（步骤3.3），并导出保存的参数（步骤3.4）。 <ol><li> 选择实验DA响应进行模拟。 <ol><li>通过输入相应文本框中的研究编号，响应数量和刺激持续时间来选择实验DA响应进行模拟。按"输入"键或单击模拟开始模拟过程，创建包含实验数据（粗绿线），模拟数据（蓝色虚线）和不考虑后刺激DA释放的模拟数据的3个图形（红色虚线）， 如图2所示 。 </li></ol></li><li> 建模实验DA响应。 注意:建模的目的是调整与激发的DA释放相关的模型参数（步骤3.2.1），DA再摄取步骤3.2.2）和刺激后DA释放（步骤3.2.3）以准确模拟实验DA响应。建模是一个迭代过程，其中开始参数被精炼，直到参数产生近似DA对时间图（A图）中的实验数据的模拟DA响应，其可以通过将模拟数据拟合到实验导出的数据来辅助估计的后刺激成分图（B组）和释放，再摄取和EO图的速率（图C）。 <ol><li>调整与DA释放相关的ΔDAR，ΔDARτ和DARss参数（ 等式2 ）以匹配刺激的幅度。 注意:这些参数只是开始估算值，而ΔDARτ= 25，DARss值是ΔDAR的1/5，这是令人满意的起始条件。增加任何这些参数将增加DA与时间图中的振幅，并增加DA相关估计的后刺激释放组件和释放，再摄取和EO图的速率。 </li><li>调整与DA再摄取（等式4）相关联的V max ，K mi ， ΔKm ，K minf和k参数，使得在图A中，模拟数据近似于实验数据的上升相位的形状线），并且使得在刺激后释放轨迹（红色虚线）上的模拟小于所有刺激后时间点的实验数据轨迹。 注意:此步骤可能需要重新调整DA释放参数（步骤3.2.1）。 </li><li>调整与刺激后DA释放（ 等式3 ）相关的XR，τR和m参数，使得仿真近似于DA对时间图中的实验数据。 注意:X R应取0到1之间的值，通常应大于0.7。 </li></ol></li><li> 保存/ LO建模参数。 <ol><li>一旦一组参数对实验数据进行了密切的模拟，单击"保存参数"，将为该项目的.mat文件保存给定响应的参数集。 </li><li>如果需要，通过单击加载参数来加载特定响应的先前保存的参数。确保在相应的文本框中输入适当的学习编号和答复编号。 </li></ol></li><li> 导出保存的参数。 <ol><li>在"导出参数"按钮旁边的文本框中，键入文件名（ 例如 Sample.txt），然后单击"导出参数"以导出具有模拟所有参数的文本文件。使用空格分隔文本文件以生成参数电子表格，如下面" 代表结果"部分的图4所示。 </li></ol></li></ol>

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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Response+times+of+carbon+fiber+microelectrodes+to+dynamic+changes+in+catecholamine+concentration.">Response times of carbon fiber microelectrodes to dynamic changes in catecholamine concentration.</a> Anal Chem. 74 (3), 539-546 (2002).</li><li>May, L. J., Wightman, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterogeneity+of+stimulated+dopamine+overflow+within+rat+striatum+as+observed+with+in+vivo+voltammetry.">Heterogeneity of stimulated dopamine overflow within rat striatum as observed with in vivo voltammetry.</a> Brain research. 487 (2), 311-320 (1989).</li><li>Wu, Q., Reith, M. E., Wightman, R. M., Kawagoe, K. T., Garris, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+release+and+uptake+parameters+from+electrically+evoked+dopamine+dynamics+measured+by+real-time+voltammetry.">Determination of release and uptake parameters from electrically evoked dopamine dynamics measured by real-time voltammetry.</a> J Neurosci Methods. 112 (2), 119-133 (2001).</li></ol>

作者没有什么可以披露的。

使用FSCV研究体内刺激的DA神经传递起源于20世纪80年代30 ，仍然是具有无与伦比的空间和时间分辨率的体内神经传递数据的丰富来源。受刺激的DA响应反映了由电刺激本身调制的DA释放和再摄取的复杂平衡。 QN模型结合了来自当代神经传递研究的原理，在动态释放和再摄取假设方面对体内刺激的DA神经传递数据进行建模。此外，QN框架将可能的FSCV分析范围...

多巴胺（DA）神经传递在各种认知和行为功能中起重要作用，其功能障碍涉及几种常见的疾病和障碍。因此，开发定量研究体内 DA神经传递的准确方法至关重要，以评估DA神经传递在疾病模型和药物药理学背景下的变化。快速扫描循环伏安法（FSCV）允许在体内 DA神经传递监测，具有良好的空间和时间分辨率。虽然可以监测生理DA神经传递在清醒，自由行动的动物，麻醉动物的上升多巴胺能途径的电刺激可以产生稳定的DA反应，适合DA神经传递的增强的动力学分析。 电刺激的DA响应反映了DA释放和再摄取以及解释的动态相互作用这些反应主要使用被称为Michaelis-Menten（MM）模型12的刺激DA神经传递的简单模型。 MM模型由3个变量组成，根据恒定的DA释放速率和恒定的再摄取效率（ 即， DA再摄取速率和细胞外DA浓度之间的关系）来描述DA响应，如等式1所述 : <img alt="公式1" src="/files/ftp_upload/55595/55595eq1.jpg" /> （DA释放）（DA再摄取） 在等式1中， f是刺激的频率; [DA] p是每脉冲刺激的估计的DA浓度增加; V max表示估计的最大再摄取速率;并且K m是估计的MM常数，其在理论上等同于饱和DAT的50％的细胞外DA浓度，导致半最大再摄取速率。这个差异可以通过估计[DA] p ，V max和K m参数来整合等式来模拟实验DA响应。 虽然MM模型有助于在各种实验环境中了解DA神经传递动力学方面的显着进步，但MM模型制作了简单的基本假设，限制了其在超生理刺激引起的DA反应建模时的适用性。例如，如果MM模型以凸起方式上升，则MM模型只能近似DA响应形状，但不能解释在背纹纹区域12中发现的渐进（凹）上升响应。因此，MM模型假设不能准确地捕获受激DA神经传递的动态释放和再摄取过程。 根据现实的量子来模拟受激DA响应基于互补研究和实验的刺激神经传递动力学原理，开发了定量神经生物学（QN）框架。各种神经传递研究表明，（1）刺激的神经递质释放是在刺激过程中速率降低的动态过程14 ，（2）在双相衰变动力学15后的刺激后期释放继续发生，和（3）DA在刺激持续时间2,16期间，再摄取效率逐渐被抑制。这三个概念作为QN框架的基础，三个方程由描述DA释放和再摄取动力学的12个参数组成（ 表1 ）。 QN框架可以密切模拟异质实验DA响应类型，以及p刺激参数和药物给药的实验操作的改变效果2,6 。虽然进一步的研究是必要的，以完善数据建模方法，未来的实验可以极大地受益于这种神经生物学接地的建模方法，这显着增加了从受刺激的DA神经传递范式中得出的推论。 <img alt="表格1" src="/files/ftp_upload/55595/55595tbl1.jpg" /> 表1: 建模方程和参数 。 <a href="http://ecsource.jove.com/files/ftp_upload/55595/55595tbl1large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> 本教程介绍如何使用QNsim 1.0对受激DA响应数据进行建模以估计DA释放和再摄取动力学。实际实验数据收集这里不描述加工，并且仅需要时间DA浓度数据。 QN框架的理论支持和基础已经在前面已经有很多的描述2 ，但是对应用QN框架来建立DA响应数据的实际观点如下所述。 QN框架模拟动态相互作用:1）动态DA释放，2）DA再摄取; 3）超生理刺激对这些过程的影响，从DA响应数据中提取有意义的动力学信息。 QN框架最适合于使用高持续时间（ 例如， 60 Hz，10 s刺激）的高度超生理刺激获取的FSCV数据，其产生适用于动力学分析的强大的DA响应。在对底层释放和再摄取过程进行准确建模之后，可以使用模型参数来模拟DA反应，其应近似于ex感应DA响应。 QN框架的方程式描述了在受刺激的DA反应过程中DA释放和再摄取的速率。当刺激的DA释放速率在刺激过程中指数下降时，QN框架描述了刺激的DA释放速率作为从刺激开始（t 刺激 ）开始的时间的函数。这与容易释放的池的耗尽一致，具有补充的稳态DA释放速率（DARss）以解释囊泡补充，类似于其他报告（ 等式2 ） 14,17 。 <img alt="公式2" src="/files/ftp_upload/55595/55595eq2.jpg" />增加DA释放速率的操作，例如增加ΔDAR，ΔDARτ或DARss导致DA与时间图的响应幅度增加。每个参数ter对DA响应形状有差异。增加DARss和ΔDARτ都使得响应的上升阶段更线性（较少凸）。降低ΔDARτ促进凸度，其由ΔDAR的大小控制。基于建模经验，DARss通常小于ΔDAR的1/5;因此，ΔDAR是主要确定DA响应的总体响应幅度的释放参数。 刺激后DA释放速率由等式3建模为刺激结束时刺激的DA释放速率（DAR ES ）的延续，作为刺激后（t post ）时间的函数。后刺激DA释放速率遵循双相衰变模式，如前所述15 ，具有快速指数衰减期和延长的线性衰变期以模拟两个ca依赖于依赖的神经递质释放过程。 <img alt="公式4" src="/files/ftp_upload/55595/55595eq4.jpg" /> （快速指数衰减）（长时间线性衰减） 目前不可能确定发生多少刺激后DA释放。可以通过系统地最小化刺激后DA释放的估计和跨越从使用不同刺激持续时间的同一记录位点收集的一组实验DA响应来验证模型参数来解决这个限制。这种最小化允许用户对释放和再摄取进行保守的估计。因为电刺激导致促进刺激后神经递质释放的钙积聚，刺激的持续时间影响后刺激神经元垃圾释放参数18,19 。基于建模经验，发现随着刺激持续时间的增加，τR增加，X R降低，与更大的钙积累的预期效果相一致20 。 方程4描述了作为MM框架的扩展的DA再摄取速率并且包含动态K m项，其在刺激期间增加以模拟由超生理刺激2,16引起的逐渐降低的再摄取效率。刺激结束后K m保持恒定在K m值（K mES ）。 <img alt="公式5" src="/files/ftp_upload/55595/55595eq5.jpg" />哪里， <img alt ="等式6"src ="/ files / ftp_upload / 55595 / 55595eq6.jpg"/&gt; （刺激期间）（刺激后） 刺激的DA反应，特别是从腹侧纹状体区域，往往对初始K m值（K mi ）的变化不敏感，这使得定义K mi值有问题。因此，像原始的MM框架一样，从对照未处理的动物12收集的DA反应，K mi近似为0.1-0.4μM。 ΔK m项决定了刺激过程中再摄取效率变化的程度，从我们的经验来看，约为2081; M在60Hz，10s刺激过程中。 k和K minf值确定K m随着时间的推移而改变，并且增加这些项中的任一个可以促进上升阶段的凹陷。 V max是部分与局部DA转运体密度相关的最大再摄取速率，其表现出腹外侧至背侧梯度21 。因此，背侧纹状体（D-Str）中的V max值通常大于30μM / s，但通常在腹侧区域（如伏核（NAc） 6 ））小于30μM / s。 上述一般准则可以帮助建模实验DA响应数据，但是生成近似实验DA响应的仿真需要迭代地调整模型参数。可以通过获得提供的超生理刺激的DA响应来改善模型参数的准确性一个强大的基板用于模拟，以及通过获得和建模多个DA响应来刺激在相同记录位置（ 例如， 60Hz，5s和10s的刺激）的不同持续时间以验证参数的准确性（参见样本数据）。为了证明，在已经使用QN框架建模的药理学挑战之前和之后，在包含在伏隔核和背纹纹状体中收集的区域特异性刺激的DA应答的软件包中包含数据集。通过扩展，用户将发现这种方法可以类似地应用于表征DA神经传递在各种疾病背景和药理学操作中的动力学。

<table><tbody><tr><td>MATLAB R2016a for Mac&amp;nbsp;</td><td>Mathworks</td><td></td><td></td></tr><tr><td>QNsim1.0</td><td>In house software package</td><td></td><td>Software to model FSCV data using the QN framework</td></tr></tbody></table>

modeling fast-scan cyclic voltammetry data from electrically stimulated dopamine neurotransmission data using qnsim1.0

中枢多巴胺能（DAergic）途径在广泛的功能（如注意力，动机和运动）中具有重要作用。多巴胺（DA）涉及疾病和病症，包括注意缺陷多动障碍，帕金森病和创伤性脑损伤。因此，DA神经传递及其研究方法具有强烈的科学兴趣。 体内快速扫描循环伏安法（FSCV）是一种允许以精细的时间和空间分辨率选择性地监测DA浓度变化的方法。这种技术通常用于上升DAergic途径的电刺激以控制多巴胺神经传递的脉冲流动。尽管受刺激的DA神经传递范例可以产生具有清晰形态学的强大的DA反应，使其适合于动力学分析，但仍然存在关于如何根据其DA释放和清除来解释反应的许多争论ce组件。为了解决这个问题，最近开发了一种刺激DA神经传递的定量神经生物学（QN）框架，以在刺激的DA反应过程中实时模拟DA释放和再摄取的动力学。该模型的基础是基于来自刺激的DA神经传递的实验数据和来自各种研究线路的神经传递原理。 QN模型实现了与激发的DA释放和再摄取动力学相关的12个参数以模拟DA响应。这项工作描述了如何使用QNsim1.0模拟DA响应，还详细说明了系统地分辨刺激的多巴胺释放和再摄取动力学变化的原理。

软件程序包括从大鼠背纹纹（研究1）和伏隔核（研究2）获得的样品DA神经传递数据，被编译成"Sample.xlsx"。电子表格包含对DA运输抑制剂哌甲酯（MPH）（10mg / kg）施用35分钟后对60Hz，10s和5s刺激的基线响应的DA浓度数据和对60Hz，5s刺激的响应。 kg，ip）。刺激的DA响应数据被组织成列， 如图3所示 ，其用作用于组织使用软件程序建模的数据的模板。 <p class="jove_content" fo:keep-tog...

Watch this Scientific Journal Video about Modeling Fast-scan Cyclic Voltammetry Data from Electrically Stimulated Dopamine Neurotransmission Data Using QNsim1.0 at JoVE.com

Modeling Fast-scan Cyclic Voltammetry Data from Electrically Stimulated Dopamine Neurotransmission Data Using QNsim1.0

快速扫描循环伏安法可以在药物，疾病和其他实验操作的上下文中监测体内多巴胺神经传递。这项工作描述了QNsim1.0的实现，QNsim1.0是一种根据定量神经生物学模型建模电刺激多巴胺反应的软件，用于量化多巴胺释放和再摄取动力学的估计。

使用QNsim1.0从电刺激多巴胺神经传递数据建模快速扫描循环伏安数据

Central dopaminergic (DAergic) pathways have an important role in a wide range of functions, such as attention, motivation, and movement. Dopamine (DA) is ...

modeling-fast-scan-cyclic-voltammetry-data-from-electrically

Research

JoVE Journal

Neuroscience

9.8K 次观看.  University of Pittsburgh. 快速扫描循环伏安法可以在药物，疾病和其他实验操作的上下文中监测体内多巴胺神经传递。这项工作描述了QNsim1.0的实现，QNsim1.0是一种根据定量神经生物学模型建模电刺激多巴胺反应的软件，用于量化多巴胺释放和再摄取动力学的估计。 

视频: Modeling Fast-scan Cyclic Voltammetry Data from Electrically Stimulated Dopamine Neurotransmission Data Using QNsim1.0

Examination of Rapid Dopamine Dynamics with Fast Scan Cyclic Voltammetry During Intra-oral Tastant Administration in Awake Rats

Rapid, phasic dopamine (DA) release in the mammalian brain plays a critical role in reward processing, reinforcement learning, and motivational control. Fast scan cyclic voltammetry (FSCV) is an electrochemical technique with high spatial and temporal (sub-second) resolution that has been utilized to examine phasic DA release in several types of preparations. In vitro experiments in single-cells and brain slices and in vivo experiments in anesthetized rodents have been used to identify mechanisms that mediate dopamine release and uptake under normal conditions and in disease models. Over the last 20 years, in vivo FSCV experiments in awake, freely moving rodents have also provided insight of dopaminergic mechanisms in reward processing and reward learning. One major advantage of the awake, freely moving preparation is the ability to examine rapid DA fluctuations that are time-locked to specific behavioral events or to reward or cue presentation. However, one limitation of combined behavior and voltammetry experiments is the difficulty of dissociating DA effects that are specific to primary rewarding or aversive stimuli from co-occurring DA fluctuations that mediate reward-directed or other motor behaviors. Here, we describe a combined method using in vivo FSCV and intra-oral infusion in an awake rat to directly investigate DA responses to oral tastants. In these experiments, oral tastants are infused directly to the palate of the rat &#8211; bypassing reward-directed behavior and voluntary drinking behavior &#8211; allowing for direct examination of DA responses to tastant stimuli.

Rapid fluctuations in extracellular dopamine (DA) mediate both reward processing and motivated behavior in mammals. This manuscript describes the combined use of fast scan cyclic voltammetry (FSCV) and intra-oral tastant administration to determine how tastants alter rapid dopamine release in awake, freely moving rats.

考试多巴胺快速与动态快速扫描循环伏安法在口内呈味物质管理清醒大鼠

Rapid, phasic dopamine (DA) release in the mammalian brain plays a critical role in reward processing, reinforcement learning, and motivational control. ...

科研

行为学

Presynaptic Dopamine Dynamics in Striatal Brain Slices with Fast-scan Cyclic Voltammetry

Extensive research has focused on the neurotransmitter dopamine because of its importance in the mechanism of action of drugs of abuse (e.g. cocaine and amphetamine), the role it plays in psychiatric illnesses (e.g. schizophrenia and Attention Deficit Hyperactivity Disorder), and its involvement in degenerative disorders like Parkinson's and Huntington's disease. Under normal physiological conditions, dopamine is known to regulate locomotor activity, cognition, learning, emotional affect, and neuroendocrine hormone secretion. One of the largest densities of dopamine neurons is within the striatum, which can be divided in two distinct neuroanatomical regions known as the nucleus accumbens and the caudate-putamen. The objective is to illustrate a general protocol for slice fast-scan cyclic voltammetry (FSCV) within the mouse striatum. FSCV is a well-defined electrochemical technique providing the opportunity to measure dopamine release and uptake in real time in discrete brain regions. Carbon fiber microelectrodes (diameter of ~ 7 &mu;m) are used in FSCV to detect dopamine oxidation. The analytical advantage of using FSCV to detect dopamine is its enhanced temporal resolution of 100 milliseconds and spatial resolution of less than ten microns, providing complementary information to in vivo microdialysis.

Using fast-scan cyclic voltammetry to measure electrically evoked presynaptic dopamine dynamics in striatal brain slices.

突触前多巴胺纹状体脑片的快速扫描循环伏安动态

Extensive research has focused on the neurotransmitter dopamine because of its importance in the mechanism of action of drugs of abuse (e.g. cocaine and ...

神经科学

Voltage Biasing, Cyclic Voltammetry, &amp; Electrical Impedance Spectroscopy for Neural Interfaces

Electrical impedance spectroscopy (EIS) and cyclic voltammetry (CV) measure properties of the electrode-tissue interface without additional invasive procedures, and can be used to monitor electrode performance over the long term. EIS measures electrical impedance at multiple frequencies, and increases in impedance indicate increased glial scar formation around the device, while cyclic voltammetry measures the charge carrying capacity of the electrode, and indicates how charge is transferred at different voltage levels. As implanted electrodes age, EIS and CV data change, and electrode sites that previously recorded spiking neurons often exhibit significantly lower efficacy for neural recording. The application of a brief voltage pulse to implanted electrode arrays, known as rejuvenation, can bring back spiking activity on otherwise silent electrode sites for a period of time. Rejuvenation alters EIS and CV, and can be monitored by these complementary methods. Typically, EIS is measured daily as an indication of the tissue response at the electrode site. If spikes are absent in a channel that previously had spikes, then CV is used to determine the charge carrying capacity of the electrode site, and rejuvenation can be applied to improve the interface efficacy. CV and EIS are then repeated to check the changes at the electrode-tissue interface, and neural recordings are collected. The overall goal of rejuvenation is to extend the functional lifetime of implanted arrays.

Voltage Biasing, Cyclic Voltammetry, & Electrical Impedance Spectroscopy for Neural Interfaces

The electrode-tissue interface of neural recording electrodes can be characterized with electrical impedance spectroscopy (EIS) and cyclic voltammetry (CV). Application of voltage biasing changes the electrochemical properties of the electrode-tissue interface and can improve recording capability. Voltage biasing, EIS, CV, and neural recordings are complementary.

偏置电压，循环伏安，神经接口电阻抗光谱

Electrical impedance spectroscopy (EIS) and cyclic voltammetry (CV) measure properties of the electrode-tissue interface without additional invasive ...

Single Cell Measurement of Dopamine Release with Simultaneous Voltage-clamp and Amperometry

After its release into the synaptic cleft, dopamine exerts its biological properties via its pre- and post-synaptic targets1. The dopamine signal is terminated by diffusion2-3, extracellular enzymes4, and membrane transporters5. The dopamine transporter, located in the peri-synaptic cleft of dopamine neurons clears the released amines through an inward dopamine flux (uptake). The dopamine transporter can also work in reverse direction to release amines from inside to outside in a process called outward transport or efflux of dopamine5. More than 20 years ago Sulzer et al. reported the dopamine transporter can operate in two modes of activity: forward (uptake) and reverse (efflux)5. The neurotransmitter released via efflux through the transporter can move a large amount of dopamine to the extracellular space, and has been shown to play a major regulatory role in extracellular dopamine homeostasis6. Here we describe how simultaneous patch clamp and amperometry recording can be used to measure released dopamine via the efflux mechanism with millisecond time resolution when the membrane potential is controlled. For this, whole-cell current and oxidative (amperometric) signals are measured simultaneously using an Axopatch 200B amplifier (Molecular Devices, with a low-pass Bessel filter set at 1,000 Hz for whole-cell current recording). For amperometry recording a carbon fiber electrode is connected to a second amplifier (Axopatch 200B) and is placed adjacent to the plasma membrane and held at +700 mV. The whole-cell and oxidative (amperometric) currents can be recorded and the current-voltage relationship can be generated using a voltage step protocol. Unlike the usual amperometric calibration, which requires conversion to concentration, the current is reported directly without considering the effective volume7. Thus, the resulting data represent a lower limit to dopamine efflux because some transmitter is lost to the bulk solution.

The amperometric technique measures dopamine release from a single cell by detecting the oxidative current produced by spontaneous dopamine oxidization. Simultaneous voltage clamp and amperometry methodology reveal the mechanistic relationship between the overall "activity" of dopamine transporter and the regulatory role of this activity on the reverse transport of dopamine.

单细胞的多巴胺释放，同时电压钳和电流分析法测量

After its release into the synaptic cleft, dopamine exerts its biological properties via its pre- and post-synaptic targets1. The dopamine signal is ...

Gold Nanoparticle Modified Carbon Fiber Microelectrodes for Enhanced Neurochemical Detection

For over 30 years, carbon-fiber microelectrodes (CFMEs) have been the standard for neurotransmitter detection. Generally, carbon fibers are aspirated into glass capillaries, pulled to a fine taper, and then sealed using an epoxy to create electrode materials that are used for fast scan cyclic voltammetry testing. The use of bare CFMEs has several limitations, though. First and foremost, the carbon fiber contains mostly basal plane carbon, which has a relatively low surface area and yields lower sensitivities than other nanomaterials. Furthermore, the graphitic carbon is limited by its temporal resolution, and its relatively low conductivity. Lastly, neurochemicals and macromolecules have been known to foul at the surface of carbon electrodes where they form non-conductive polymers that block further neurotransmitter adsorption. For this study, we modify CFMEs with gold nanoparticles to enhance neurochemical testing with fast scan cyclic voltammetry. Au3+ was electrodeposited or dipcoated from a colloidal solution onto the surface of CFMEs. Since gold is a stable and relatively inert metal, it is an ideal electrode material for analytical measurements of neurochemicals. Gold nanoparticle modified (AuNP-CFMEs) had a stability to dopamine response for over 4 h. Moreover, AuNP-CFMEs exhibit an increased sensitivity (higher peak oxidative current of the cyclic voltammograms) and faster electron transfer kinetics (lower &#916;EP or peak separation) than bare unmodified CFMEs. The development of AuNP-CFMEs provides the creation of novel electrochemical sensors for detecting fast changes in dopamine concentration and other neurochemicals at lower limits of detection. This work has vast applications for the enhancement of neurochemical measurements. The generation of gold nanoparticle modified CFMEs will be vitally important for the development of novel electrode sensors to detect neurotransmitters in vivo in rodent and other models to study neurochemical effects of drug abuse, depression, stroke, ischemia, and other behavioral and disease states.

In this study, we modify carbon-fiber microelectrodes with gold nanoparticles to enhance the sensitivity of neurotransmitter detection.

用于增强神经化学检测的金纳米粒子改性碳纤维微电极

For over 30 years, carbon-fiber microelectrodes (CFMEs) have been the standard for neurotransmitter detection. Generally, carbon fibers are aspirated into ...

化学

Dopamine Release at Individual Presynaptic Terminals Visualized with FFNs

The nervous system transmits signals between neurons via neurotransmitter release during synaptic vesicle fusion. To observe neurotransmitter uptake and release from individual presynaptic terminals directly, we designed fluorescent false neurotransmitters as substrates for the synaptic vesicle monoamine transporter. Using these probes to image dopamine release in the striatum, we made several observations pertinent to synaptic plasticity. We found that the fraction of synaptic vesicles releasing neurotransmitter per stimulus was dependent on the stimulus frequency. A kinetically distinct "reserve" synaptic vesicle population was not observed under these experimental conditions. A frequency-dependent heterogeneity of presynaptic terminals was revealed that was dependent in part on D2 dopamine receptors, indicating a mechanism for frequency-dependent coding of presynaptic selection.

Hui Zhang and Niko G. Gubernator contributed equally to this work.

A new means to measure neurotransmission optically using fluorescent dopamine analogs.

多巴胺的释放突触前货箱码头在个别的可视化与FFNs

The nervous system transmits signals between neurons via neurotransmitter release during synaptic vesicle fusion. To observe neurotransmitter uptake and ...

生物学

Measuring In Vivo Changes in Extracellular Neurotransmitters During Naturally Rewarding Behaviors in Female Syrian Hamsters

The ability to measure neurotransmitter release on a rapid time scale allows patterns of neurotransmission to be linked to specific behaviors or manipulations; a powerful tool in elucidating underlying mechanisms and circuitry. While the technique of microdialysis has been used for decades to measure nearly any analyte of interest in the brain, this technique is limited in temporal resolution. Alternatively, fast scan cyclic voltammetry is both temporally precise and extremely sensitive; however, because this technically difficult method relies on the electroactivity of the analyte of interest, the possibility to detect nonelectroactive substances (e.g., the neurotransmitter glutamate) is eliminated. This paper details the use of a turn-key system that combines fixed-potential amperometry and enzymatic biosensing to measure both electroactive and nonelectroactive neurotransmitters with temporal precision. The pairing of these two powerful techniques allows for the measurement of both tonic and phasic neurotransmission with relative ease, and permits recording of multiple neurotransmitters simultaneously. The aim of this manuscript is to demonstrate the process of measuring dopamine and glutamate neurotransmission in vivo using a naturally rewarding behavior (i.e., sexual behavior) in female hamsters, with the ultimate goal of displaying the technical feasibility of this assay for examining other behaviors and experimental paradigms.

Measuring In Vivo Changes in Extracellular Neurotransmitters During Naturally Rewarding Behaviors in Female Syrian Hamsters

This paper details the use of fixed-potential amperometric recordings using carbon fiber electrodes and enzymatic biosensor technology to measure the release of dopamine and glutamate with high temporal resolution during natural rewarding behavior in the female hamster.

雌性叙利亚仓鼠自然奖赏行为中细胞外神经递质的测量体内变化

The ability to measure neurotransmitter release on a rapid time scale allows patterns of neurotransmission to be linked to specific behaviors or ...

Combined Infusion and Stimulation with Fast-Scan Cyclic Voltammetry (CIS-FSCV) to Assess Ventral Tegmental Area Receptor Regulation of Phasic Dopamine

Phasic dopamine (DA) release from the ventral tegmental area (VTA) to the nucleus accumbens plays a pivotal role in reward processing and reinforcement learning. Understanding how the diverse neuronal inputs into the VTA control phasic DA release can provide a better picture of the circuitry that controls reward processing and reinforcement learning. Here, we describe a method that combines intra-VTA cannula infusions of pharmacological agonists and antagonists with stimulation-evoked phasic DA release (combined infusion and stimulation, or CIS) as measured by in vivo fast-scan cyclic voltammetry (FSCV). Using CIS-FSCV in anesthetized rats, a phasic DA response can be evoked by electrically stimulating the VTA with a bipolar electrode fitted with a cannula while recording in the nucleus accumbens core. Pharmacological agonists or antagonists can be infused directly at the stimulation site to investigate specific VTA receptors&#39; roles in driving phasic DA release. A major benefit of CIS-FSCV is that VTA receptor function can be studied in vivo, building on in vitro studies.

Combined Infusion and Stimulation with Fast-Scan Cyclic Voltammetry CIS-FSCV to Assess Ventral Tegmental Area Receptor Regulation of Phasic Dopamine

The goal of this protocol is to directly manipulate ventral tegmental area receptors to study their contribution to subsecond dopamine release.

联合输注和刺激与快速扫描循环伏安法（CIS-FSCV）评估腹侧被盖区域受体对相位多巴胺的调节

Phasic dopamine (DA) release from the ventral tegmental area (VTA) to the nucleus accumbens plays a pivotal role in reward processing and reinforcement ...

Cyclic Voltammetry (CV)

Source: Laboratory of Dr. Kayla Green &#8212; Texas Christian University
A Cyclic Voltammetry (CV) experiment involves the scan of a range of potential voltages while measuring current. In the CV experiment, the potential of an immersed, stationary electrode is scanned from a predetermined starting potential to a final value (called the switching potential) and then the reverse scan is obtained. This gives a &#39;cyclic&#39; sweep of potentials and the current vs. potential curve derived from the data is called a cyclic voltammogram. The first sweep is called the &#39;forward scan&#39; and the return wave is called the &#39;reverse scan&#39;. The potential extremes are termed the &#39;scan window&#39;. The magnitude of reduction and oxidation currents and the shape of the voltammograms are highly dependent on analyte concentration, scan rates, and experimental conditions. By varying these factors, cyclic voltammetry can yield information regarding the stability of transition metal oxidation state in the complexed form, reversibility of electron transfer reactions, and information regarding reactivity. This video will explain the basic setup for a cyclic voltammetry experiment including analyte preparation and setting up the electrochemical cell. A simple cyclic voltammetry experiment will be presented.

Cyclic Voltammetry CV: Measuring Redox Potentials and Currents

循环伏安法 (CV)

Source: Laboratory of Dr. Kayla Green &#8212; Texas Christian University
A Cyclic Voltammetry (CV) experiment involves the scan of a range of potential ...

教育

JoVE Science Education

分析化学

Monitoring Dopamine Release in a Rat Model with VTA Dopaminergic Neuron Receptor Manipulation

Source: Wickham, R. J., et al. <a href="https://app.jove.com/v/60886">Combined Infusion and Stimulation with Fast-Scan Cyclic Voltammetry (CIS-FSCV) to Assess Ventral Tegmental Area Receptor Regulation of Phasic Dopamine.</a> J. Vis. Exp. (2020).
This video demonstrates the implantation of electrodes and cannulas in an anesthetized rat to measure dopamine release in the nucleus accumbens using a potentiostat and the infusion of drugs to modulate dopaminergic activity in the ventral tegmental area.

使用 VTA 多巴胺能神经元受体作监测大鼠模型中的多巴胺释放

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
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