我们感谢王晔讨论和行为训练索林Pojoga。由美国国立卫生研究院尤里卡计划，国家眼科研究所，皮尤学者奖励计划，詹姆斯S ·麦克唐纳基金会（VD），以及美国国立卫生研究院视力训练津贴（BJH）支持。

1。南Microdrive微型建设南电极驱动系统相结合，我们使用的U型探头。建立这个系统需要2-3个小时，但一旦建成，这是非常简单的修改。我们开始组装，其中包括一个4通道的基础（图2a）南塔，南腔（图2b），网格间距1毫米（图2c），1-4螺钉微型硬盘（图2d），1 -4导管（图2E，直径为500微米，并削减约5-7厘米），和1-4微型硬盘塔（图2F）。为方便起见，我们将描述为建设有一个塔和一个U - Probe公司南系统过程。一些训练后，此过程通常需要2-3个小时，如果所有材料都可用。 <ol><li>要兴建南电极驱动器组装，第一项措施是大会的所有工具和您需要的件（如导管，导丝，完整dremil设置，南工具和零件和U型探头）。测量导管连接到录音设备时，他们足够长的时间不损害硬脑膜上休息。 </li><li>要兴建南电极驱动器组装，第一项措施是录音室的深度。然后，切断与测量长度约5-7厘米的导管。在切割导管，需要，以确保没有金属碎片进入管内。使用硬线小于直径的导管内管内删除任何金属碎片。 </li><li>下一步，放置到南基地南电网。拧紧的紧固螺钉和电网螺杆。一旦基地和电网的安全，确定录制地区利益，并事先通过导管南电网的底部。 </li><li>通过电网的，直到它是南腔外约1-2毫米的导管。一旦导管在所需的位置，开始组装南Microdrive微型硬盘塔。 </li><li>南Microdrive微型硬盘塔在每个有两个夹子 - 电机驱动器的顶部钳位，可以是固定到位或松动而底部钳的。将顶部钳的钢筋管的U - Probe公司。底部钳将导管，适用于少量的强力胶固定到位导管。这个系统更稳定和更精确的加强U型探头管连接到两个夹子。 </li><li>小心地将导管顶部的U型探头的尖端，并通过导管U型探头，直到你可以安全的塔南基地。塔的位置与翼形螺钉调整，U型探针或导管，以便有没有增加紧张。 </li></ol> 2。 U型探头消毒该层的电极或Plextrode U型探头是从Plexon公司公司购买，并在2000美元左右的价格 - $ 4000。价格取决于三个主要方面:联系站点，站点配置，每个站点的直径。目前，我们正在使用的线性配置16通道版本和一个直径为25微米的接触。更重要的是，U型探头的厚度直接相关的接触直径。在我们的实验中，我们一直使用直径为25微米的接触，这是等于360微米的厚度。目前我们的版本模式的成本约为3500美元。 U型探头都装在一个电极的情况下跳线和接地线和所需的时间从购买到交付大约4-6周。 <ol><li>气缸的基础上南系统和电机电缆连接到相应的塔。颜色编码压缩的关系，如果使用多个塔是用来帮助区分电机电缆和塔。 </li><li>使用南软件程序，开始推进U型探头，可以设置目标位置，自动向前推进的U型探头到该位置，或通过点击""南软件接口。推进U型探针，使至少10毫米的提示，通过过去南腔导管。 </li><li>消毒U型探针，MetriCide活性双醛溶液20-30分钟植入的录音室之前，连接南基地。之后，用无菌水冲洗U型探头和NaN基地。 </li><li> U型探头，提示导管内零收回南软件的位置。在南软件中，单击"零各个岗位。 </li><li>将南基地植入的录音室，并拧紧所有四个螺丝。然后，根据录音室的一侧是一个引脚对齐基地。所有四个螺丝拧紧，确保南基地是牢固地连接到录音室。 </li></ol> 3。提前录制的U型探头鉴于杜拉的强度和厚度，主体之间是高度可变的，我们已经实施了推进的U - PRO的一般程序使用南Microdrive微型硬盘系统。更重要的是，每个U型探头的详细分析的每一个接触阻抗和护林员为U - Probe公司的整体。我们用电极的接触阻抗从0.3-0.5MΩ不等。目前有阻抗测试仪从Plexon公司购买的，但不幸的是，在我们的录音时，此设备不可用。因此，我们已无法执行了详细的阻抗分析。 <ol><li> U型探头是左浮动（有一根导线连接在底部连接器的跳线）。的探头固定在U型探头连接器和放大器的电缆连接和接地。 </li><li>约1-2毫米的初始发展既要快而强。设置速度参数范围在0.1 - 0.2毫米/秒，并深入一步，以0.2 - 0.3毫米。这些值将确保U型探头能够穿破硬脊膜干净，并在录音中是一个重要的第一步。 </li><li>一旦通过硬脑膜，减少0.050 -0.1毫米/秒的速度，并降低了深入一步，以0.05 - 0.1毫米。我们的目标是推进顺利，尽可能等缓慢，无组织被破坏的U - Probe公司。探头进入大脑的迹象之一是变化幅度在减少噪音水平（文字叠加:局部领域的潜力）的陪同下LFP。 </li><li>为了验证电极是跨越所有皮质层，测量振幅在满场的白色闪光刺激变化。跨越时间在大幅面幅度的变化，诱发反应的潜力分析的基础。此分析提供确定皮质层的基础上。 </li></ol> 4。皮质层的识别和验证我们有皮质层，使用诱发反应电位（ERP）范式和电流源密度​​（CSD）分析确定的程序实施。我们依赖于惩教署，因为它提供了一个索引的位置，方向，和跨膜电流流过密度，使我们能够准确定位电极记录在一个单一的渗透层。事实上，查尔斯施罗德和他的同事们先前结合层记录，microlesion和组织学的重建，以验证在9-12在V1的皮质层的功能鉴定ERP / CSD方法的有效性。使用自发产生的振荡其他方法已用于鉴别皮质的深度，如皮质纱锭和向上/向下国家13-15。 基于上述分析，我们利用MATLAB中，计算惩教署根据第二LFP跨越U型探头的间距相等的接触时间序列的空间衍生（ <a href="http://software.incf.org/software/csdplotter/home">ICSD</a>工具箱http://software.incf.org/ <a href="http://software.incf.org/software/csdplotter/home">软件/ csdplotter /家庭</a> ）9,10,16,17。 <ol><li>要确定的皮质层，测量过程中处于被动的固定任务的诱发反应电位，同时露出了一个满场的黑色屏幕，闪烁白色为100毫秒，然后返回到黑色。构成此序列是重复200次试验。 </li><li> Plexon公司的多通道采集处理器保存所有的连续数据信号直接录制的计算机通过NI的PCI板卡。保存数据后，开始处理的信号电流源密度​​分析。 </li><li>使用的软件校正FPAlign Plexon公司提供正确的过滤器中的探头和前置放大板诱导的磷酸锂铁信号的时间延迟。 </li><li>在这一点上的数据转移到MATLAB与Neuroexplorer。每个LFP通道过滤使用标准高和低通滤波器的截止频率为0.5赫兹和100赫兹。每个电极接触已筛选后，确定审判和平均每跨试验，以获得每个电极接触的时间序列的平均劳动力参与。然后，整理成一个矩阵作为时间的函数的振幅与LFP每个联系人。 </li><li>运行ICSD（文本覆盖密度电流源）在MATLAB工具箱在工作区输入CSDplotter。鉴于连续数据采样频率为1 kHz，DT参数设置为1毫秒。接下来，设置皮层的电导率值0.4 S / M（约在每立方毫米毫微安单位电流源密度​​），改变电极作为[0.1:0.1:1.6的矢量位置，以反映接触。当所有参数都被插入，单击"运行"。 </li><li>查看惩教署在CSDplotter接口配置文件，并将其粘贴到一个新的数字。在MATLAB中的常见功能，如imagesc可用于积层剖面，可以代表惩教署的数据和比较小时层识别应用和各种平滑算法和规范化程序和会话。 </li><li>要识别极性反转源库在第4层的基础配置的陪同下，首先，验证，在颗粒层使用层惩教署的档案的主要下沉。找到驱动负极在惩教署图水槽。然后，计算颗粒下沉的质量中心。 </li><li>是从一个质心的联络电话号码和时间组成的分析时，接收器是最大的。与接收器质心的接触作为0微米的颗粒层参考。分析分为三个层参考和集团他们的上方和下方的所有联系人:supragranular，粒状，和infragranular。 </li><li>验证洗牌的电极位置，离开时域不变的颗粒下沉。惩教署矩阵洗牌后，重新计算质心的分析。洗牌作为皮质深度功能的电极接触，应销毁任何层的特殊性。 </li></ol> 5。单个神经元的鉴定和接受实地测绘我们曾与隔离和记录多个单单位，从U - Probe公司巨大的成功。在一个典型的记录，我们可以期望有6-10隔离的单位和14-16局部领域的潜力信号。寻找单一的单位也是单电极相比，与U - Probe公司更可靠。即使使用所有必要的硬件，能够精确地提前16个电极，他们将无法探索，正是与U - Probe公司的皮质层功能的网络人口。最后，我们通常可以记录与相同的U型探头为30-40穿透。 <ol><li>为了找到接受的领域，开始呈现反向相关的刺激感受野有可能位于显示器。由四个方向光栅0，45，90和135度的刺激。 </li><li>执行聚类分析射速地图找到的感受野。首先，计算每个时间延迟的最大射速的位置和它们的质心。然后，计算质心和这些最大射速地点之间的距离。计算40之间的传导延​​迟发射率在每个空间位置的地图 - 120毫秒，每个神经元在5毫秒的间隔独立。 </li><li>查找在所有时间延迟之间的质心与周围的最大射速点的总距离。感受野的时间延迟最小化，距离。 </li><li>一旦发现一个感受野是为每一个细胞，目前反向相关刺激比感受野纪录的人口重叠的感受野的位置较大。一个实时的发射率情节，可以用来判断是否正确的感受野位置已经确定。 </li><li>最后，删除单个单位突然改变他们的反应，只保留单位作进一步的分析与稳定射击率。此外，选择了最佳的信号噪声比的记录点。 </li></ol> 6。代表结果:单单位的录音从初级视觉皮层和皮质层之间的LFPs 在使用电极层的分析最重要的步骤之一是能够可靠地确定皮质层，并验证在许多时间和会话识别。因此，我们在全领域闪现刺激（图3a）测量的跨层接触LFPs诱发反应电位（ERPs）。图3b提供的信息类型之一，需要获得以计算电流源密度​​（CSD），以确定皮质层的一个例子。然后，我们惩教署LFP时间序列分析，以确定极性反转，伴随着在第4层的基础下沉源配置。图4a所示的惩教署在整个皮质的深度本地​​化作为时间函数的皮质层的分析 - supragranular（SG），颗粒（G）和infragranular（IG）层保持稳定甚至四个小时后，记录会话开始的位置。图4b包括惩教署的痕迹，代表平均分配到一个给定的的图层这些接触 - 在这个例子中，颗粒层经历了一个明显减少幅度在惩教署〜50毫秒。这种分析作为参考分配supragranular和infragranular层颗粒层上方和下方的电极接触，分别为（最大下沉0微米的颗粒层参考服务中心的质量与接触）。 另一个关键的分析，使用层电极准确地识别和本地化的神经元的感受野。此过程是对定位的刺激产生的最强有力的反应，从神经元是至关重要的。图5a是两个神经元的感受野地块的初级视觉CORT例如EX（V1）。这些地块的起源是固定点，这是一个黑色的电脑屏幕上集中显示一个白色的小圆圈。在这些地块的颜色代表每个神经元的放电率，在一个动态的反向相关性刺激的反应。我们利用这些信息来定位为一个给定的实验（如正弦波光栅）的刺激。比一般的感受野大小，呈现的刺激较大，以涵盖所​​有同时记录神经元的感受野位置。 后，我们确定了皮质层，位置最佳的感受野位置的刺激，我们可以进行动物进行实验的协议中，我们提出不同的视觉刺激，而无论是固定或歧视的任务。实验后，我们执行我们的尖峰波形分析，隔离单单位，我们能够记录相同的通道。这一过程往往需要一些时间，掌握和不断改进新的分析软件和技术提供。图5b是一个使用Plexon公司的离线浏览"后，人们可能预期的输出类型的例子。使用这个软件，单一的单元之间的隔离是通过目视检查。确定不同的集群基础上的第一和第二主成分的重量，穗宽，山谷，和峰值性能。 <img src="/files/ftp_upload/2806/2806fig1.jpg" alt="图1" /> 图1。多接触层电极 ，使用多接触电极层，我们录得同时扣球从孤立的单个神经元的活动和跨大幅面单位V1的皮质层。每个U型探头由16等距（100微米）的电极接触，涵盖了总长度为1.6毫米。每个电极的接触是25微米，直径铂铱组成。 <img src="/files/ftp_upload/2806/2806fig2.jpg" alt="图2" /> 图2。南电网建设南Microdrive微型硬盘系统提供额外的古典螺杆驱动的微型硬盘的稳定性和精度。每个电极组是独立操纵在XY平面，一个用户定义的工作范围内。每个电极组在Z方向独立操纵在一个用户定义的工作深度（可达100 mm）和变速范围从0.001毫米/秒至0.5毫米/秒和1微米的高分辨率（一 ）4。 -通道的基础，（二）南腔，（C），间距1毫米，（四 ）1-4螺钉微型硬盘，（五 ）1-4导管（直径为500微米，并削减约5-7厘米）电网（F）1-4微型硬盘塔和（七）完成的南系统和气缸基地。 <img src="/files/ftp_upload/2806/2806fig3.jpg" alt="图3" /> 图3。诱发反应潜在的范式和LFP时间系列（一 ）确定皮质层中，我们测量的诱发反应电位（ERP）在被动的固定任务，而猴子们接触到一个满场的黑色屏幕上闪现白（〜1Hz的100）毫秒，然后返回到黑色（B）LFP层U型探头记录的反应处理，以获得ERP系统为每个联系人的痕迹。颗粒层中的所有会议决定，由定位一个接收器驱动的反转，并在ERP痕迹的响应幅度的存在伴随着在第4层的基础下沉源配置的极性反转。反转发生时，虚线框表示一段时间的时间。 <img src="/files/ftp_upload/2806/2806fig4.jpg" alt="图4" /> 图4。层识别，使用电流源密度分析（一 ）电流源密度分析（第二LFP时间序列的空间衍生）被用来确定伴随着在基片源配置的极性反转颗粒层。我们评估如何皮质层的识别是随着时间的推移保持稳定（从左至右）。在这些例子中，当前接收器（蓝色）代表颗粒层和跨越〜400微米（二）惩教署下面的每一个情节的痕迹代表平均分配到一个给定的的图层这些接触的惩教署。这使我们能够确定的初始下沉的精确定时（在这些例子〜50-60毫秒。惩教署跟踪信封代表标准偏差和黑网吧表明闪过刺激的持续时间（100毫秒）。 <img src="/files/ftp_upload/2806/2806fig5.jpg" alt="图5" /> 图5。尖峰排序和接受实地测绘（一）首先，一个可视化程度的一半计算了一倍。然后，反向相关性刺激的CRT显示器C补丁面向光栅onsisting 0，45，90和135度。每个神经元的发放率是40至120毫秒之间的时间间隔在5毫秒计算独立后，刺激每个空间位置。最大发射率的计算，然后每次延迟的质心。然后，在每个延迟的质心与相邻的射速地点之间的距离计算。选择时间延迟的最小距离是感受野（二）使用脱机分拣软件程序穗波形的属性，如峰高，河谷深度，高峰，谷时间，峰值或谷等进行了分析（ Plexon公司）。尖峰基于类似物业进行排序，直到从一个神经元的波形没有从另一个重叠集群。 <img src="/files/ftp_upload/2806/2806fig6.jpg" alt="图6" /> 图6。洗牌惩教署剖面图3a相同的约定，但我们进行了一个洗牌的过程，随机混合与接触地点编译一个新的惩教署矩阵。这种分析是用来更好地验证洗牌的电极位置，离开时域不变的颗粒下沉。从这些例子显示随着时间的推移，洗牌作为皮质深度功能的电极接触会破坏任何层的特殊性。

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多单元的录音已经成为分析皮层中的神经网络是如何刺激信息进行编码的标准。执行层电极由于电极技术的最新进展，使局部皮层电路前所未有的特性。虽然多电极记录提供了有关神经种群动态的有用信息，使多个层电极更高的分辨率和更多的神经元的具体位置信息。由于皮层与解剖不同的输入和输出层的组织，这引起了在这些层disparately感觉信息是如何处理的问题。 我们已经提出了一个新的记录方法，利用多接触层的电极记录在初级视觉皮层（V1），皮质层的功能本地网络的活动。重要的是，我们还实施了分析过程中诱发的响应模式的局部领域的潜力，以确定皮质层的方法。我们还提供了从接受实地测绘程序和尖峰波形分析的详细结果。 我们承认，不是没有限制，最引人注目的是录制的稳定层电极。我们奉劝那些使用这种技术，耐心地推进电极，并允许的时间足够数量的大脑后进步定居（我们通常记录45分钟至1小时后，最后提前）。在这段时间内，我们将运行无数眼球校准，接受映射，和诱发反应的潜在范例。 我们能够提高我们的录音，使用导管固定用螺丝微型硬盘，这是基地的南。我们还修改了标准的U型探头的设计，减少30到25度，刀尖角度。因此，U型探头的幅度，允许通过硬脑膜顺畅渗透。有可能与钝电极尖端的组织损伤和出血可能发生。出血可覆盖的电极接触，防止清洁单元之间的隔离。我们已经测试了这一理论的录音和30日和25度的针尖角度和能够解决更多的单位超过穿透，甚至延长生命的U型探头。 如前所述以上我们通常提前开始，并迅速减慢，一旦我们通过硬脑膜通过。我们相信，这个程序结合锐利的刀尖角度，使我们成为一个能够解决单个单位活动，使用U - Probe公司的几个实验室。我们能使大脑定居后的U - Probe公司地位的时间长短直接关系到我们的单个单元活动和录制的整体稳定。 这项技术将继续蓬勃发展，随着越来越多的实验室将利用这些技术。目前，长期植入阵列的设计和实施工作正在进行，将最有可能取代多电极电网。此外，含有多个沿轴（实质上多个U型探针）接触电极的阵列，正在开发并行。

<table border="1"><tbody><tr><td> 设备名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>南微硬盘系统</td><td>南仪器</td><td>南S4 </td><td>图2。定制的夹具需要使用U - Probe公司。 U型探头异常提到的一切是由南仪器。 </td></tr><tr><td>螺丝微型硬盘</td><td> MIT机械厂</td><td></td><td>凡是能够争取到一个导管南电网应是适当的。 </td></tr><tr><td>不锈钢导管</td><td>小零件</td><td> B00137QHNS（1）或B00137QHO2（5） </td><td>这些都是在长60，并在实验室使用DREMEL手电钻切割的大小</td></tr><tr><td> Plexon公司ü探头</td><td> Plexon公司，公司</td><td> PLX - UP - 16 - 25ED - 100 - SE - 360 - 25T - 500 </td><td> U型探头规格请<a href="www.plexon.com">www.plexon.com</a>见图1。 </td></tr></tbody></table> 表1。五金。 <table border="1"><tbody><tr><td> 软件名称 </td><td> 公司 </td><td> 网站 </td><td> 评论 </td></tr><tr><td>南软件</td><td>南</td><td> <a href="http://www.naninstruments.com/DesignConcept.htm">http://www.naninstruments.com/DesignConcept.htm</a> </td><td>计算机接口需要一个额外的的串行端口，以适应Plexon公司系统及南硬件</td></tr><tr><td>离线浏览"，FPAlign，PlexUtil，MATLAB程序</td><td> Plexon公司</td><td> <a href="http://www.plexon.com/downloads.html#Software">http://www.plexon.com/downloads.html＃软件</a> </td><td>在"安装软件包" </td></tr><tr><td> NeuroExplorer </td><td> NeuroExplorer </td><td> <a href="http://www.neuroexplorer.com/">http://www.neuroexplorer.com/</a> </td><td>在"资源" </td></tr><tr><td> CSDplotter版本0.1.1 </td><td>克拉斯阁下Petterson </td><td> <a href="http://arken.umb.no/~klaspe/user_guide.pdf">http://arken.umb.no/〜klaspe / user_guide.pdf</a> </td><td></td></tr></tbody></table> 表2。软件。

examining local network processing using multi-contact laminar electrode recording

皮质层是无处不在整个大脑皮层1-4高度经常性的本地网络的组成结构。近年来，已取得了重大进展，我们的理解差异，在应对不同的皮质层5-8神经元属性，但仍然是一个伟大的左了解神经元的人群是否，以及如何在特定层的信息进行编码处理方式。 现有的多电极阵列技术，虽然测量皮质空间沿皮层表面的许多毫米的反应信息，是不适宜的做法层皮质电路问题。在这里，我们提出我们的方法，为建立和记录单个神经元和跨越的初级视觉皮层（V1）皮质层局部场电位（LFPs），利用多接触电极层（图1; Plextrode U型探头，Plexon公司公司）。 的方法包括录音设备建设，皮质层的识别，并确定单个神经元的感受野。要找出皮质层，我们衡量LFP使用满场闪现刺激时间序列诱发反应电位（ERPs）。然后，我们执行的电流源密度（CSD）分析，以确定在基地的第4层（第4层的水槽内，后来被称为颗粒层 9-12）片源配置的陪同下极性反转。电流源密度是有用的，因为它提供了一个索引的位置，方向，和跨膜电流流过密度，使我们能够准确定位电极记录在一个单一的渗透层6，11，12。

Watch this Scientific Journal Video about Examining Local Network Processing using Multi-contact Laminar Electrode Recording at JoVE.com

Examining Local Network Processing using Multi-contact Laminar Electrode Recording

我们了解皮质电路中的一个根本的问题是，如何在不同的皮质层的网络感官的信息进行编码。在这里，我们描述了利用多接触层电极的电生理技术记录单单位和地方领域的潜力和目前的分析，以确定皮质层。

检查本地网络处理采用多接触层电极记录

Cortical layers are ubiquitous structures throughout neocortex1-4 that consist of highly recurrent local networks. In recent years, significant progress ...

examining-local-network-processing-using-multi-contact-laminar

Nanyang Technological University

Research

JoVE Journal

Neuroscience

12.7K 次观看.  University of Texas. 我们了解皮质电路中的一个根本的问题是，如何在不同的皮质层的网络感官的信息进行编码。在这里，我们描述了利用多接触层电极的电生理技术记录单单位和地方领域的潜力和目前的分析，以确定皮质层。

视频: Examining Local Network Processing using Multi-contact Laminar Electrode Recording

Correlating Behavioral Responses to fMRI Signals from Human Prefrontal Cortex: Examining Cognitive Processes Using Task Analysis

The aim of this methods paper is to describe how to implement a neuroimaging technique to examine complementary brain processes engaged by two similar tasks. Participants' behavior during task performance in an fMRI scanner can then be correlated to the brain activity using the blood-oxygen-level-dependent signal. We measure behavior to be able to sort correct trials, where the subject performed the task correctly and then be able to examine the brain signals related to correct performance. Conversely, if subjects do not perform the task correctly, and these trials are included in the same analysis with the correct trials we would introduce trials that were not only for correct performance. Thus, in many cases these errors can be used themselves to then correlate brain activity to them. We describe two complementary tasks that are used in our lab to examine the brain during suppression of an automatic responses: the stroop1 and anti-saccade tasks. The emotional stroop paradigm instructs participants to either report the superimposed emotional 'word' across the affective faces or the facial 'expressions' of the face stimuli1,2. When the word and the facial expression refer to different emotions, a conflict between what must be said and what is automatically read occurs. The participant has to resolve the conflict between two simultaneously competing processes of word reading and facial expression. Our urge to read out a word leads to strong 'stimulus-response (SR)' associations; hence inhibiting these strong SR's is difficult and participants are prone to making errors. Overcoming this conflict and directing attention away from the face or the word requires the subject to inhibit bottom up processes which typically directs attention to the more salient stimulus. Similarly, in the anti-saccade task3,4,5,6, where an instruction cue is used to direct only attention to a peripheral stimulus location but then the eye movement is made to the mirror opposite position. Yet again we measure behavior by recording the eye movements of participants which allows for the sorting of the behavioral responses into correct and error trials7 which then can be correlated to brain activity. Neuroimaging now allows researchers to measure different behaviors of correct and error trials that are indicative of different cognitive processes and pinpoint the different neural networks involved.

The goal of our research is to correlate behavior to brain activity. Accurate behavioral measures and imaging techniques allow us to elucidate brain-behavior relationships.

关联从人类前额叶皮层的fMRI信号的行为反应:检查的认知过程，使用任务分析

The aim of this methods paper is to describe how to implement a neuroimaging technique to examine complementary brain processes engaged by two similar ...

科研

神经科学

Electrophysiology on Isolated Brainstem-spinal Cord Preparations from Newborn Rodents Allows Neural Respiratory Network Output Recording

While it is well known that the central respiratory drive is located in the brainstem, several aspects of its basic function, development, and response to stimuli remain to be fully understood. To overcome the difficulty of accessing the brainstem in the whole animal, isolation of the brainstem and part of the spinal cord is performed. This preparation is maintained in artificial cerebro-spinal fluid where gases, concentrations, and temperature are controlled and monitored. The output signal from the respiratory network is recorded by a suction electrode placed on the fourth ventral root. In this manner, stimuli can be directly applied onto the brainstem, and the effect can be recorded directly. The signal recorded is linked to the inspiratory signal sent to the diaphragm via the phrenic nerve, and can be described as bursts (around 8 bursts per minute). Analysis of these bursts (frequency, amplitude, length, and area under the curve) allows precise characterization of the stimulus effect on the respiratory network. The main limitation of this method is the viability of the preparation beyond the early post-natal stages. Thus, this method greatly focuses on the study of the whole network without the peripheral inputs in the newborn rat.

The central respiratory drive is located in the brainstem. Spontaneous respiratory motor output from an isolated brainstem-spinal cord is recorded by placing an electrode on the fourth ventral root. This experimental approach is valuable for pharmacological investigations or the assessment of respiratory challenges and genetic manipulations on rhythmic motor behavior.

电从新生儿鼠害隔离脑干，脊髓准备允许呼吸中枢网络输出记录

While it is well known that the central respiratory drive is located in the brainstem, several aspects of its basic function, development, and response to ...

Direct-current Stimulation and Multi-electrode Array Recording of Seizure-like Activity in Mice Brain Slice Preparation

Cathodal transcranial direct-current stimulation (tDCS) induces suppressive effects on drug-resistant seizures. To perform effective actions, the stimulation parameters (e.g., orientation, field strength, and stimulation duration) need to be examined in mice brain slice preparations. Testing and arranging the orientation of the electrode relative to the position of the mice brain slice are feasible. The present method preserves the thalamocingulate pathway to evaluate the effect of DCS on anterior cingulate cortex seizure-like activities. The results of the multichannel array recordings indicated that cathodal DCS significantly decreased the amplitude of the stimulation-evoked responses and duration of 4-aminopyridine and bicuculline-induced seizure-like activity. This study also found that cathodal DCS applications at 15 min caused long-term depression in the thalamocingulate pathway. The present study investigates the effects of DCS on thalamocingulate synaptic plasticity and acute seizure-like activities. The current procedure can test the optimal stimulation parameters including orientation, field strength, and stimulation duration in an in vitro mouse model. Also, the method can evaluate the effects of DCS on cortical seizure-like activities at both the cellular and network levels.

Studies have shown that cathodal transcranial direct-current stimulation can produce suppressive effects on drug-resistant seizures. In this study, an in vitro experimental setup was devised in which the direct-current stimulation and multielectrode array recording of seizure-like activity were evaluated in mice brain slice preparation. The direct-current stimulation parameters were evaluated.

小鼠脑切片准备扣押样活性的直接电流刺激和多电极阵列记录

Cathodal transcranial direct-current stimulation (tDCS) induces suppressive effects on drug-resistant seizures. To perform effective actions, the ...

Time-dependent Increase in the Network Response to the Stimulation of Neuronal Cell Cultures on Micro-electrode Arrays

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network dynamics in neuronal cultures. In contrast with more traditional methods, such as patch-clamping, which can only record activity from a single cell, MEAs can record simultaneously from multiple sites in a network, without requiring the arduous task of placing each electrode individually. Moreover, numerous control and stimulation configurations can be easily applied within the same experimental setup, allowing for a broad range of dynamics to be explored. One of the key dynamics of interest in these in vitro studies has been the extent to which cultured networks display properties indicative of learning. Mouse neuronal cells cultured on MEAs display an increase in response following training induced by electrical stimulation. This protocol demonstrates how to culture neuronal cells on MEAs; successfully record from over 95% of the plated dishes; establish a protocol to train the networks to respond to patterns of stimulation; and sort, plot, and interpret the results from such experiments. The use of a proprietary system for stimulating and recording neuronal cultures is demonstrated. Software packages are also used to sort neuronal units. A custom-designed graphical user interface is used to visualize post-stimulus time histograms, inter-burst intervals, and burst duration, as well as to compare the cellular response to stimulation before and after a training protocol. Finally, representative results and future directions of this research effort are discussed.

Mouse neuronal cells cultured on multi-electrode arrays display an increase in response following electrical stimulation. This protocol demonstrates how to culture neurons, how to record activity, and how to establish a protocol to train the networks to respond to patterns of stimulation.

时间依赖性增加对微电极阵列刺激神经元细胞培养物的网络响应

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network ...

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 ...

Examining Monosynaptic Connections in Drosophila Using Tetrodotoxin Resistant Sodium Channels

Here, a new technique termed Tetrotoxin (TTX) Engineered Resistance for Probing Synapses (TERPS) is applied to test for monosynaptic connections between target neurons. The method relies on co-expression of a transgenic activator with the tetrodotoxin-resistant sodium channel, NaChBac, in a specific presynaptic neuron. Connections with putative post-synaptic partners are determined by whole-cell recordings in the presence of TTX, which blocks electrical activity in neurons that do not express NaChBac. This approach can be modified to work with any activator or calcium imaging as a reporter of connections. TERPS adds to the growing set of tools available for determining connectivity within networks. However, TERPS is unique in that it also reliably reports bulk or volume transmission and spillover transmission.

Examining Monosynaptic Connections in Drosophila Using Tetrodotoxin Resistant Sodium Channels

This article features a method to test the monosynaptic connections between neurons by employing tetrodotoxin and the tetrodotoxin-resistant sodium channel, NaChBac.

使用河豚毒素耐药性钠通道检查果蝇中的 Monosynaptic 连接

Here, a new technique termed Tetrotoxin (TTX) Engineered Resistance for Probing Synapses (TERPS) is applied to test for monosynaptic connections between ...

Evaluation of Hemisphere Lateralization with Bilateral Local Field Potential Recording in Secondary Motor Cortex of Mice

This article demonstrates complete, detailed procedures for both in vivo bilateral recording and analysis of local field potential (LFP) in the cortical areas of mice, which are useful for evaluating possible laterality deficits, as well as for assessing brain connectivity and coupling of neural network activities in rodents. The pathological mechanisms underlying Alzheimer&#39;s disease (AD), a common neurodegenerative disease, remain largely unknown. Altered brain laterality has been demonstrated in aging people, but whether or not abnormal lateralization is one of the early signs of AD has not been determined. To investigate this, we recorded bilateral LFPs in 3-5-month-old AD model mice, APP/PS1, together with littermate wild type (WT) controls. The LFPs of the left and right secondary motor cortex (M2), specifically in the gamma band, were more synchronized in APP/PS1 mice than in WT controls, suggesting a declined hemispheric asymmetry of bilateral M2 in this AD mouse model. Notably, the recording and data analysis processes are flexible and easy to carry out, and can also be applied to other brain pathways when conducting experiments that focus on neuronal circuits.

We present in vivo electrophysiological recording of the local field potential (LFP) in bilateral secondary motor cortex (M2) of mice, which can be applied to evaluate hemisphere lateralization. The study revealed altered levels of synchronization between the left and right M2 in APP/PS1 mice compared to WT controls.

小鼠二次运动皮质中双边局部场电位记录半球横向化评价

This article demonstrates complete, detailed procedures for both in vivo bilateral recording and analysis of local field potential (LFP) in the cortical ...

Advanced Diffusion Imaging in The Hippocampus of Rats with Mild Traumatic Brain Injury

Mild traumatic brain injury (mTBI) is the most common type of acquired brain injury. Since patients with traumatic brain injury show a tremendous variability and heterogeneity (age, gender, type of trauma, other possible pathologies, etc.), animal models play a key role in unraveling factors that are limitations in clinical research. They provide a standardized and controlled setting to investigate the biological mechanisms of injury and repair following TBI. However, not all animal models mimic the diffuse and subtle nature of mTBI effectively. For example, the commonly used controlled cortical impact (CCI) and lateral fluid percussion injury (LFPI) models make use of a craniotomy to expose the brain and induce widespread focal trauma, which are not commonly seen in mTBI. Therefore, these experimental models are not valid to mimic mTBI. Thus, an appropriate model should be used to investigate mTBI. The Marmarou weight drop model for rats induces similar microstructural alterations and cognitive impairments as seen in patients who sustain mild trauma; therefore, this model was selected for this protocol. Conventional computed tomography and magnetic resonance imaging (MRI) scans commonly show no damage following a mild injury, because mTBI induces often only subtle and diffuse injuries. With diffusion weighted MRI, it is possible to investigate microstructural properties of brain tissue, which can provide more insight into the microscopic alterations following mild trauma. Therefore, the goal of this study is to obtain quantitative information of a selected region-of-interest (i.e., hippocampus) to follow up disease progression after obtaining a mild and diffuse brain injury.

The overall goal of this procedure is to obtain quantitative microstructural information of the hippocampus in a rat with mild traumatic brain injury. This is done using an advanced diffusion-weighted magnetic resonance imaging protocol and region-of-interest based analysis of parametric diffusion maps.

轻度创伤性脑损伤大鼠海马学的高级扩散成像

Mild traumatic brain injury (mTBI) is the most common type of acquired brain injury. Since patients with traumatic brain injury show a tremendous ...

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

Estimation of the number of dopaminergic neurons in the substantia nigra is a key method in pre-clinical Parkinson's disease research. Currently, unbiased stereological counting is the standard for quantification of these cells, but it remains a laborious and time-consuming process, which may not be feasible for all projects. Here, we describe the use of an image analysis platform, which can accurately estimate the quantity of labeled cells in a pre-defined region of interest. We describe a step-by-step protocol for this method of analysis in rat brain and demonstrate it can identify a significant reduction in tyrosine hydroxylase positive neurons due to expression of mutant &#945;-synuclein in the substantia nigra. We validated this methodology by comparing with results obtained by unbiased stereology. Taken together, this method provides a time-efficient and accurate process for detecting changes in dopaminergic neuron number, and thus is suitable for efficient determination of the effect of interventions on cell survival.

Here we present an automated method for semi-quantitative determination of dopaminergic neuron number in the rat substantia nigra pars compacta.

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 ...

Recording Network Activity in Spinal Nociceptive Circuits Using Microelectrode Arrays

The roles and connectivity of specific types of neurons within the spinal cord dorsal horn (DH) are being delineated at a rapid rate to provide an increasingly detailed view of the circuits underpinning spinal pain processing. However, the effects of these connections for broader network activity in the DH remain less well understood because most studies focus on the activity of single neurons and small microcircuits. Alternatively, the use of microelectrode arrays (MEAs), which can monitor electrical activity across many cells, provides high spatial and temporal resolution of neural activity. Here, the use of MEAs with mouse spinal cord slices to study DH activity induced by chemically stimulating DH circuits with 4-aminopyridine (4-AP) is described. The resulting rhythmic activity is restricted to the superficial DH, stable over time, blocked by tetrodotoxin, and can be investigated in different slice orientations. Together, this preparation provides a platform to investigate DH circuit activity in tissue from na&#239;ve animals, animal models of chronic pain, and mice with genetically altered nociceptive function. Furthermore, MEA recordings in 4-AP-stimulated spinal cord slices can be used as a rapid screening tool to assess the capacity of novel antinociceptive compounds to disrupt activity in the spinal cord DH.

The combined use of microelectrode array technology and 4-aminopyridine-induced chemical stimulation for investigating network-level nociceptive activity in the spinal cord dorsal horn is outlined.