我们感谢德意志民主共和国（DFG），KFO 247，为我们的研究提供资金。

实验程序按照德国"动物福利法"（2014年最后修订）和欧洲条例（2010/63 / EU）进行。实验由当地动物福利机构（LaGeSo，Berlin）批准，符合当地部门和国际准则。 注意:在所提出的方法中，使用两个模型的电极从连接主运动皮质（M1）与底丘核（STN）和黑质基质（SNr）连接的超直接皮质基底神经节途径记录。对于使用M1定制低阻抗Ag / AgCl电极的硬膜外皮质电图（ECoG）记录。来自STN和SNr的记录用市售的高阻钨电极进行。 硬膜外Ag / AgCl硬膜外电极的构建<ol><li>拿一个约5厘米长的99.99％纯银线条，带直径r为200μm，如有必要，可以去除任何涂层。 </li><li>将电极尖端向下插入打火机或蜡烛的火焰中，直到尖端开始熔化。等到尖端是球形，直径约为1毫米。将预制的电极从球形尖端的开始到导线端部的总长度为15mm。 </li><li>焊接精密连接器到线端，适合所用的电生理记录系统。用导电银漆覆盖焊丝端到连接器的焊点。这有助于电导率，并产生更好的信号质量。 </li><li>导电清漆干燥后，用3 mm至1 mm的热收缩管覆盖焊点。仔细使用制表师的锤子将球形尖端压平至厚度的一半。 </li><li>戴上检查手套，取100％乙醇的无绒清洁布清除任何污垢和油脂。 </li><li>把电极放入15 mL离心管或细胞培养管，并用家用氯漂白剂（注意:每100g溶剂含有2.8克次氯酸钠），直到球形尖端完全被覆盖。 注意:氯漂白剂有腐蚀性;始终遵循制造商的安全说明。 </li><li> 23分钟后取出电极，用蒸馏水漱口。氯化银层的成功应用表现为均匀的紫色变化。 </li><li>在空气中干燥完全干燥后，用细镊子取电极。使用精美的画笔，应用液体电气绝缘。直接从电极尖端的电线上开始，并覆盖所有的热收缩管。让绝缘干燥至少2小时。 </li><li>为了进行质量控制，请用万用表检查电导率。如果可用，使用适当的阻抗表在1 kHz下执行阻抗测试，同时进行电极和测试将探针放在含有0.9％NaCl的H 2 O溶液中而不相互接触。 1 kHz时的阻抗值应为约8kΩ。 </li></ol> 2.将电极固定在立体定位架上注意:要同时记录MUA和LFP，请使用阻抗为1.5MΩ的钨微丝电极。如果记录的焦点在单个单元的高质量记录上，请选择具有较高阻抗（&gt; 5MΩ）的微线电极。如果研究的目的仅针对LFP，则具有较低阻抗的电极是可以接受的。对于经常需要进行背侧立体定位调整的小型结构，可以使用具有合适的背侧尖端分离（在这种情况下为250μm）的成对电极。此外，如果需要，这提供了更局部的参考电极的优点。始终从最下面的电极和a测量立体定位坐标重新计算参考伯尔贾。 <ol><li>拿一个带有丙烯酸块和夹具的标准立体定位电极支架，并将其牢牢放在手术显微镜视野的平坦表面上。 </li><li>使用细镊子将第一对电极松散地固定在带有胶带（3毫米×8毫米）的支架的丙烯酸块上。电极应突出丙烯酸块约12毫米。 </li><li>仔细地将第二双极电极固定在第一电极的旁边。为了瞄准超直接路径的结构，距离应为2mm（ 图1 ）。对于大多数标准的立体定位电极支架，这是相邻的凹槽。使用卡尺来验证。可以以相同的方式接近不同的网络。为此，可以将丙烯酸嵌段转变到一定程度。 </li><li>通过将第二对电极小心地滑动到最接近顶端约200的位置来调整第二对电极与第一电极相比凹陷（ 图1 ）。在微观视觉下进行。为此，使用30 G插管（外径300μm）来更好地估计距离。 </li><li>按下胶带，然后用夹具的金属夹固定。 </li></ol>手术<ol><li>对于电生理记录，使用聚氨酯（小心）进行麻醉。 注意:氨基甲酸是有毒和致癌的，因此始终遵守物质制造商提供的安全规定和数据表。 </li><li>在0.9％的NaCl食盐水溶液中制备200 mg / mL聚氨酯溶液。 </li><li>腹膜内（IP）总共施用1.3g / kg体重尿布。根据大鼠应变，在注射之间以15分钟间隔将剂量分成两个剂量可能是合理的，以便增强麻醉的安全性。 </li><li>使用p检查麻醉深度戒烟反射和其他合适的反射。麻醉不足以进行手术，注射0.15 g / kg体重的尿烷IP，再等15分钟。 </li><li>应用眼膏防止角膜脱水。 </li><li>在麻醉期间不断监测呼吸频率和踏板撤退反射。使用温度控制的小型动物加热垫，以确保在整个手术过程中保持生理体温。在开始电生理记录之前，更换为非电气替代品（ 如醋酸钠头垫）。 </li><li>剃毛与头部的背侧，以实现一个干净的手术领域。用适当的手术消毒剂在切口部位周围消毒。将动物固定在立体定向框架中。 </li><li>用手术刀在矢状方向进行2厘米长的头皮切口。使用手术刀稍微刮掉头骨腱膜并消毒颅骨。使用公司浸泡在3％H 2 O 2中的豆芽以除去任何剩余的组织。 </li><li>如有必要，请使用电动机或电热器控制出血。停止颅骨和皮下注射出血，如果出血不能在1-2分钟后自发停止，并阻止颅骨的视线。 </li><li>调整门牙杆，直到头部位于平坦的颅骨位置，这意味着作为立体定位参考点的母体和lambda位于同一平面。这是实现高手术精度最重要的。使用标准的立体定位大鼠对齐工具，在微观视觉下校准指定的尖端至胭脂，并调整切牙杆，直到工具上的指定点和工具上的lambda同时触及颅骨。 注意:从一侧的视图与来自另一侧的聚焦光可能有助于确定这种情况。或者，取一个具有精细插管的立体定位支架，并测量Bregma a的背向坐标ndλ在微观视觉下。调整门牙杆，直到Bregma和lambda的背侧坐标相同。 </li><li>使用具有插管的立体定位夹具，校准到母体，然后计算颅骨上所有钻孔的位置。使用立体定位支架，通过仔细抓颅骨或使用手术颜色标记来标记待钻孔的位置。这个坐标取决于目标，参考表格的坐标给出了表1中的超直接路径，包括小脑参考电极的建议坐标。 </li><li>使用微型钻头仔细钻孔。对于STN和SNr，钻一个公共孔（大小为2 mm x 3 mm）。所有其他钻孔应具有约1mm的直径。 </li><li>取两个细管（至少27 G），并弯曲其尖端以形成钩形，使用硬表面或镊子。使用这些来删除任何debris从钻孔中，并在普通STN / SNr孔中仔细切割并去除硬脑膜。 </li><li>用生理盐水冲洗钻孔。每15分钟应用一滴生理盐水给钻孔，以防止脑和硬脑膜变干。 </li><li>用微型钻头和匹配的不锈钢微型螺钉（ 如 M 1.2 mm x 2 mm螺丝），在小脑上方的参考硬膜外电极的钻孔之间钻一个孔并拧入微螺钉，对于M1硬膜外电极</li><li>将自建的Ag / AgCl硬膜外电极滑入参考电极和M1电极的钻孔中。用精细的镊子引导电极尖端，并将其直接滑入颅骨下方进入钻孔。 </li><li>用双组分牙科丙烯酸修复所有硬膜外电极。确保不要覆盖边缘点，也不影响常见的STN / SNr孔。 </li><li>用钨微丝插入准备好的支架进入立体定位框架。 </li><li>校准旨在瞄准STN的大多数腹侧电极到正前方。调整到公共STN / SNr孔以上的计算位置，并在微观视觉下将电极降低到大脑。确保钨微丝电极顺利进入大脑。 </li></ol> 4.电生理测绘与录音注意:对于此步骤，需要具有能够进行在线过滤和在线穗分选的记录软件的法拉第笼和多通道电生理记录系统。优选使用与位于动物头部附近的前置放大器一起工作的系统，以将电噪声和伪像保持在绝对最小值。除了钨微丝电极之外，还需要至少一个硬膜外和一个参比电极来执行超直接通路的记录。建议插入硬膜外和参考e电极成对，没有它们相互接触，这有助于在发生故障的情况下，并允许在数据分析中进行不同类型的参考。 <ol><li>将移动法拉第笼放在立体定位框架上方。如果只有一个固定的法拉第笼可用，请仔细将立体定位框架移动到法拉第笼中，同时确保深脑电极不会下降到脑中，直到立体定位框架处于最终位置。 </li><li>将电极连接到电生理设置的前台。确保参考电极连接到适当的参考通道。 </li><li>设置录音软件:带通滤波器（0.05-8,000 Hz）并放大（增益1,500-2,000x）原始数据信号。使用适当设置的在线LFP和尖峰滤波器（LFP的带通滤波器为0.05-250 Hz，MUA为300-8,000 Hz的带通滤波器）。对于所有过滤器，请使用butterworth型过滤器。 </li><li>为onlin设置峰值阈值（如果适用）e尖峰排序。大多数记录软件允许设置尖峰阈值，其是幅度值，高于该幅度信号由软件标记为尖峰。该阈值可以以数学方式确定为滤波的尖峰信号的平均幅度的因子或标准偏差，或者可以优选地通过目视检查&lt;500ms的数据段来确定，并且在图形中设置为高于信号噪声的线用户界面。 注意:设置尖峰阈值的意图是计数尖峰和排序单位，以提供有关目前记录多少神经元以及它们的尖峰形状的信息。 </li><li>将钨微丝电极慢慢地降低到目标的背面1mm，这是超直接通路的STN。如有必要，请等待信号稳定。 </li><li>对于电生理映射，以100微米的步长将电极推向腹侧。在每一步，评估射击模式，射击r吃的和形状的钉。与图2中给出的典型示例进行比较。通常，密集的核在几个背部步骤中显示出快速连续的尖峰，而富含纤维的结构在随后的腹侧步骤中显示低的燃烧速率和较小的同质刺激活性。 </li><li>对于超直接通路，确保最内侧电极位于STN内。 注意:当检测到Zona incerta腹侧的MUA显着增加时，达到STN。腹部到STN，由于电极已经到达内囊，几乎完全停止。当大多数腹侧电极处于STN中时，钨微电极的结构确保后部，第二电极处于SNr。在STN和SNr同时记录典型的MUA可能需要以小增量进行背心式微调。注意，MUA的频率取决于实际记录的神经元数量和脑水平激活。 </li><li>一旦电极处于所需的结构，设置在线过滤和尖峰分选（ 见图4 ），然后开始记录数据。可以在LFP记录中识别的不同皮层同步状态的典型示例如图3所示。 </li></ol> 5.实验结束<ol><li>录制完成后，将电极慢慢地从脑中提起，并立即用生理盐水冲洗。电极可以在彻底冲洗和目视检查后重新使用。放电弯曲电极。 </li><li>通过IP注射过量的尿烷（2.5g / kg体重）来安乐死动物。 注:聚氨酯只能用于最终程序。 </li><li>如果需要对电极位置进行组织学验证或其他组织学染色手术，请从头骨中取出大脑并进行处理组织适当。 注意:根据预期的染色方法，可能需要经心灌注。对于电极位置的验尸验证，在大多数情况下，标准Nissl染色足以在例如冠状脑切片中观察电极轨迹。促进组织学目标验证的其它方法包括通过在电极插入16,17之前通过记录电极施加电流或应用生物相容性染料，将电诱导的损伤用于脑组织。 </li></ol>

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作者没有什么可以披露的。

在本研究中，该方法证明了如何使用连接M1与啮齿动物的STN和SNr的超直接皮层 - 基底神经节途径的实例同时从给定网络的多个位点记录细胞外电生理信号。 记录小皮质结构如STN的关键步骤是将记录电极精确地引导到靶中。在所提出的方法中，注意两个关键步骤确保了目标的高准确性。在将电极引入大脑之前，在立体定向装置中制备动物时，绝对必须确保将头骨带入"平头颅骨"位置22 。为了获得平坦的颅骨位置，立体定向框架的门牙杆的位置改变，直到粗糙和高度为参考点o颅骨在同一个背神经平面上21 。只有通过确保这个位置，立体定向地图集上的坐标可以以高水平的精度应用于个体实验室动物，因为图谱基于平坦的颅骨位置21 。此外，实验证据证明，使用个体化的平头颅骨位置的瞄准精度优于门牙杆23的固定调整。记录电极在背面平面中的位置应通过不断注册多单元活动进行微调。沿着电极轨迹的不同的核和白质结构示出了特征火焰图案（ 图2 ），其可用于重新调整电极9,20的位置。 所提出的方法的另一个重要步骤是放置r参考电极。在所提出的方案中，选择小脑皮层上方的位置，因为在这一点上，参考电极不检测作为研究中心点的皮层基底神经节活动。在对易受体积传导影响的分析方法感兴趣的研究中，应该优先考虑更多的本地参考。 聚氨酯是广泛使用的麻醉剂，用于记录动物研究中的神经元细胞外电位11,18,24,25,26。这样做的原因是，与其他麻醉剂相比，单剂量的氨基甲酸酯可以产生8-12小时的稳定且持久的麻醉，其中仅具有有限的中枢神经系统活性抑制。然而，氨基甲酸乙酯麻醉a也激活交感神经系统，这可能会导致不必要的副作用， 如高血糖27 。由于其持久的作用和缺乏拮抗其麻醉作用的有效药物，氨基甲酸酯不应用于分开数小时或数天的重复实验。如果计划在同一动物上进行多次记录会话，或者如果技术原因不使用氨基甲酸乙酯，则用异氟醚和氯胺酮和赛拉嗪等药物进行气体麻醉可以是电生理体内实验的合理替代方法28 ， 29 。这些麻醉机制的缺点是，由于半衰期短，随着药物的积累，需要比使用氨基甲酸酯更频繁的监测和调整。此外，有证据表明氨基甲酸酯可能与生理b有较小的干扰<html>雨活动比其他麻醉药30 。 本文中所有详细的记录条件都会严格确定如何进一步处理和离线分析获取的数据，因此必须根据计划的分析步骤的要求调整所有设置。由于有多种分析多通道细胞外记录的选项，因此使用可用的开源工具箱可能是有利的31 。 体内细胞外电位的记录是提供独特的时间和空间分辨率的脑信号的方法，其优于诸如功能磁共振成像和钙成像的替代方法5 。所提出的方法不仅可以应用于超直接途径的记录，而且可以容易地调整到各种其他实验模型和研究问题然而，由于它涉及到立体定向手术，因此有许多研究设置不能应用于哪里，并且应该选择非侵入性方法。 在未来，应该实现所提出的细胞外多位点记录技术与光遗传工具的组合，以进一步增强我们对不同神经精神疾病的网络功能障碍的理解，以便找到新的治疗方法。

最近关于不同神经精神障碍如帕金森病（PD）和精神分裂症的累积证据强烈地表明，他们的病理生理学是基于经常涉及皮层和皮质下结构1,2,3的延长的神经元回路的关键功能障碍。根据这一理论，疾病的临床表现是由于细胞网络的信息处理能力受损，而不是单个细胞或特定神经元素1,2,3的结果 。为了增强对这种复杂的神经精神疾病组的了解，并找到新的治疗方案，必须非常详细地描述人类患者和动物模型中无序网络的神经元动力学。优秀研究生物科目大规模网络的方法是细胞外电位的多位点电生理记录4 。使用这种方法，可以同时评估局部场电位（LFP），其主要表示由突触前电位5产生的兴奋性和抑制性突触后电流和多单位活性（MUA）的时间总和。细胞外电位的记录与研究网络的其他方法（ 例如功能性磁共振成像和钙成像）相比具有几个优点，因为它提供了更高的时间和空间分辨率，并且因为它不依赖于遗传工程生物体5 。然而，细胞外体内记录的使用是有限的，因为它是不能普遍应用的侵入性技术。 体内电生理记录可以在清醒和麻醉的动物中执行排序6 。这两种方法都伴随着具体的利弊。清醒动物的研究允许在执行定义的行为任务期间记录大脑信号，但容易发生运动相关和其他工件7,8 。另一方面，麻醉动物的记录提供了在高度定义的皮层同步状态下用最少的人造物评估LFP和MUA的机会，但是结果在一定程度上也与清醒对象9,10,11中可以发现的有所不同。 近年来，已经证明，LFP的抽样对于描绘网络活动的病理变化特别有用。其中一个突出的例子是人类患者PD的病理生理学研究和动物模型，其中可以显示皮质基底节神经节循环中增强的β振荡与帕金森症运动症状12,13相关 。作为这一研究的结果，目前正在研究β振荡是否可以用作闭环深层脑刺激的在线反馈生物标志物14,15 。 在本研究中，提供了用氨基甲酸酯麻醉的大鼠中LFP和MUA的急性多位点体内电生理记录的详细描述。证明如何通过电生理学使用标准和定制的电极以及如何构建这些电极来完整的网络，例如超直接皮层 - 基底神经节途径。特别强调如何通过co实现基底节神经核的精确靶向将立体定向手术与MUAs的在线注册相结合。

<table><tbody><tr><td>Ag/AgCl custom epidural electrodes</td><td>Goodfellow GmbH&lt;br/&gt; D-61213 Bad Nauheim, Germany&lt;br/&gt; info@goodfellow.com</td><td>Product-ID AG005127 for 99.99% silver wire</td><td>Ag/AgCl electrodes will allow for better signal quality, but may only be used in acute experiments. Possible replacement: Stainless steel electrodes</td></tr><tr><td>Stereotaxic holder with acrylic block</td><td>David Kopf Instruments,&lt;br/&gt; 7324 Elmo Street, Tujunga, CA 91042, USA</td><td>Product ID Model 1770 Standard Electrode Holder</td><td>Make sure the acrylic block has recesses which suit the electrode setup for the desired target. Acrylic blocks can easily be modified with a file to obtain the desired configuration. Possible replacement: Self-constructed electrode holders</td></tr><tr><td>Tungsten microwire electrodes 1.5 M&amp;Omega; impedance</td><td>Microprobes.com&lt;br/&gt; 18247-D Flower Hill Way&amp;nbsp; Gaithersburg, Maryland, 20879 USA</td><td>Product-ID WE3ST31.5A5-250um</td><td>The 1.5 M&amp;Omega; is necessary to record MUA and LFP at the same time. Possible replacement: Microelectrodes of different materials can be used. The electrodes have to be straight, robust and as thin as possible.</td></tr><tr><td>Rat alignment tool</td><td>David Kopf Instruments,&lt;br/&gt; 7324 Elmo Street, Tujunga, CA 91042, USA</td><td>Product ID Model 944 Rat Alignment Tool</td><td>Allows the exact orientation of the brain to match stereotaxic atlases. Possible replacement: Stereotaxic holder with a cannula</td></tr><tr><td>Two-component dental acrylic</td><td>Associated Dental Products Ltd.&lt;br/&gt; Kemdent Works, Purton, Swindon&lt;br/&gt; Wiltshire, SN5 4HT, United Kingdom</td><td>Simplex Rapid Powder Clear 225g, Product code: ACR803; Simplex Rapid Liquid 150ml, Product code: ACR920</td><td>Depending in the electrodes used, superglue might be an easy alternative, if the electrodes are small and lightweight. Possible replacement: Superglue (Cyanacrylate-based)</td></tr><tr><td>Faraday cage</td><td>Self-construction</td><td></td><td>A proper Faraday cage will be the best protection from electromagnetic artifacts, but everything which can be formed into a box shape or applied to a frame and is made of conductive material may help. Possible replacement: Aluminum foil or copper mesh</td></tr><tr><td>Electrophysiological setup with recording software and online spike-sorting capabilities</td><td>OmniPlex&lt;sup&gt;&amp;reg;&lt;/sup&gt; Neural Data Acquisition System&lt;br/&gt; Plexon Inc&lt;br/&gt; 6500 Greenville Avenue, Suite 700&lt;br/&gt; Dallas, Texas 75206&lt;br/&gt; USA</td><td></td><td>Offline sorting software is a potential alternative, multiple scripts and softwares can be found for free in the open source community.</td></tr></tbody></table>

acute in vivo electrophysiological recordings of local field potentials and multi-unit activity from the hyperdirect pathway in anesthetized rats

融合证据表明，许多神经精神疾病应被理解为大规模神经元网络的紊乱。为了更好地了解这些疾病的病理生理基础，有必要精确地表征电路的不同神经元部分之间的信息处理在哪种方式的干扰。使用细胞外体内电生理记录，可以精确地描绘神经元网络内的神经元活动。该方法的应用与替代技术相比具有几个优点， 例如功能磁共振成像和钙成像，因为它允许独特的时间和空间分辨率，并且不依赖于遗传工程生物体。然而，细胞外体内记录的使用是有限的，因为它是不能普遍应用的侵入性技术。在本文中，介绍一种简单易用的方法可以在网络的多个位置同时记录细胞外电位，例如局部场电位和多单元活动。详细说明如何使用立体定向手术和多单位记录的在线分析来实现皮质下核的精确定位。因此，证明了在麻醉动物体内如何研究完整的网络如超直肠皮质基底神经节环。

使用本文使用的记录电极，可以从STN和SNr从初级运动皮层，下丘脑核和黑质分布样本LFPs和网状细胞。最初，LFP和多单元活动一起记录在宽带信号中。此后，LFP和MUA被带通滤波器分离（LFP为0.05-250Hz，MUA为300-4,000Hz）。 为了正确定位皮质下核，特别是小结构如STN，有利的是将计划的立体坐标与在线记录的MUA信号对齐。对于可以记录STN特征MUA图案的电极轨迹（ 图2 ）9,20。 对于后续步骤的分析，它是通常需要通过主成分分析从多单元活动中定义单个单位（ 图4 ）。 在M1的LFP记录中，可以识别两个自发交替的皮质同步状态:激活状态（AS）和慢波活动（SWA）状态（ 图3 ） 18,19 。虽然SWA状态由大约1Hz的高振幅慢振荡控制，但AS的特征在于具有较低振幅的较快振荡（ 图3 ）。 <img alt="图1" class="xfigimg" src="/files/ftp_upload/55940/55940fig1.jpg" /> 图1:在标准立体定位架中设置深脑微丝电极。注意A，电极对之间的尖端分离STN和B，左侧方向的SNr电极对约为200μm左右，前后方向约2毫米。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55940/55940fig1large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> <img alt="图2" class="xfigimg" src="/files/ftp_upload/55940/55940fig2.jpg" /> 图2:针对STN的背部电极轨迹的特征多单元活动。 （ A ）腹侧后内侧丘脑核（VPM），晕a（ZI），丘脑下核（STN）和黑质黑质（SNr）的多单位记录。 VPM表现出稀疏和不规则间隔的高幅度尖峰。当接近ZI时，这种尖峰模式就停止了。当电极进入STN时，具有中等幅度的短脉冲串的典型高频点火模式ude可以观察。 SNr可以通过其高振幅和规则的发射模式来识别。 （ B）叠加在大鼠立体定向图谱21上的STN轨迹。上部:冠状面。下部:矢状面。注意电极尖端通过VPM和ZI。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55940/55940fig2large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> <img alt="图3" class="xfigimg" src="/files/ftp_upload/55940/55940fig3.jpg" /> 图3.聚氨酯麻醉期间主运动皮质LFP记录中的皮层同步状态。 （ A ）主要运动皮质代表600 s LFP记录。具有对应于激活状态（i）的高频，低幅度活动的时间段和具有较慢节奏和高的时间段<html>可以区分对应于慢波活动状态（ii）的呃振幅。 （ B ）对应的时间 - 频率图，间隔为600秒，说明（A）中呈现的LFP的0-20 Hz相对功率。暖色调表示较高的相对功率。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55940/55940fig3large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> <img alt="图4" class="xfigimg" src="/files/ftp_upload/55940/55940fig4.jpg" /> 图4:从STN多单元活动中单个单元的排序。 （ A ）主成分分析后特征空间中单位群集的三维视图。每个集群表示一个推定的单个单元。 （ B ）对应于（A）中的簇的尖峰波形和尖峰波形平均值。0fig4large.jpg"target ="_ blank"&gt;请点击此处查看此图的较大版本。 <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td> 来自Bregma的坐标 </td><td> STN </td><td> 网状部 </td><td> M1 </td><td> 参考文献1 </td><td> 参考文献2 </td></tr><tr><td>前 - 后</td><td> -3.6 </td><td> -4.8 </td><td> +3.0 </td><td> -10.0 </td><td> -10.0 </td></tr><tr><td>内侧 - 外侧</td><td> +2.5 </td><td> +2.5 </td><td> +3.0 </td><td> +3.0 </td><td> -3.0 </td></tr><tr><td>背腹</td><td> -8.0 </td><td>呐</td><td>呐</td><td>呐</td><td>呐</td></tr></tbody></table> 表1:用于记录超直流Co的立体坐标基底神经节途径。所有点均以颅骨上的标准参考点测量，单位为mm;不适用。

Watch this Scientific Journal Video about Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats at JoVE.com

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

急性<em&gt;体内</em&gt;麻醉大鼠超导途径局部场电位和多单位活动的电生理记录

在本研究中，介绍了如何在尿烷麻醉下从超直接途径进行多位点体内电生理记录的方法。

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the ...

acute-vivo-electrophysiological-recordings-local-field-potentials

Universidad San Sebastian

Research

JoVE Journal

Neuroscience

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

15.5K 次观看.  Charité University Medicine Berlin. 在本研究中，介绍了如何在尿烷麻醉下从超直接途径进行多位点体内电生理记录的方法。 

视频: Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer scale. However, current methods mostly rely on tethered cable recordings or only use unidirectional systems, allowing either recording or stimulation of neural activity, but not at the same time or same target. Here, a new wireless, bidirectional device for simultaneous multichannel recording and stimulation of neural activity in freely behaving rats is described. The system operates through a single portable head stage that both transmits recorded activity and can be targeted in real-time for brain stimulation using a telemetry-based multichannel software. The head stage is equipped with a preamplifier and a rechargeable battery, allowing stable long-term recordings or stimulation for up to 1 h. Importantly, the head stage is compact, weighs 12 g (including battery) and thus has minimal impact on the animal&#180;s behavioral repertoire, making the method applicable to a broad set of behavioral tasks. Moreover, the method has the major advantage that the effect of brain stimulation on neural activity and behavior can be measured simultaneously, providing a tool to assess the causal relationships between specific brain activation patterns and behavior. This feature makes the method particularly valuable for the field of deep brain stimulation, allowing precise assessment, monitoring, and adjustment of stimulation parameters during long-term behavioral experiments. The applicability of the system has been validated using the inferior colliculus as a model structure.

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

A wireless, bidirectional system for multi-channel neural recordings and stimulation in freely behaving rats is introduced. The system is light and compact, thus having minimal impact on the animal&#180;s behavioral repertoire. Moreover, this bidirectional system provides a sophisticated tool to assess causal relationships between brain activation patterns and behavior.

用于体内的无线双向接口记录和刺激自由行为大鼠的神经活动

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer ...

科研

行为学

Simultaneous Recordings of Cortical Local Field Potentials and Electrocorticograms in Response to Nociceptive Laser Stimuli from Freely Moving Rats

Electrocortical responses, elicited by laser heat pulses that selectively activate nociceptive free nerve endings, are widely used in many animal and human studies to investigate the cortical processing of nociceptive information. These laser-evoked brain potentials (LEPs) consist of several transient responses that are time-locked to the onset of laser stimuli. However, the functional properties of the LEP responses are still largely unknown, due to the lack of a sampling technique that can simultaneously record neural activities at the surface of the cortex (i.e., electrocorticogram [ECoG] and scalp electroencephalogram [scalp EEG]) and inside the brain (i.e., local field potential [LFP]). To address this issue, we present here an animal protocol using freely moving rats. This protocol is composed of three main procedures: (1) animal preparation and surgical procedures, (2) a simultaneous recording of ECoG and LFP in response to nociceptive laser stimuli, and (3) data analysis and feature extraction. Specifically, with the help of a 3D-printed protective shell, both ECoG and LFP electrodes implanted on the rat&#39;s skull were securely held together. During data collection, laser pulses were delivered on the rat&#39;s forepaws through gaps in the bottom of the chamber when the animal was in spontaneous stillness. Ongoing white noise was played to avoid the activation of the auditory system by the laser-generated ultrasounds. As a consequence, only nociceptive responses were selectively recorded. Using the standard analytical procedures (e.g., band-pass filtering, epoch extraction, and baseline correction) to extract stimulus-related brain responses, we obtained results showing that LEPs with a high signal-to-noise ratio were simultaneously recorded from ECoG and LFP electrodes. This methodology makes the simultaneous recording of ECoG and LFP activities possible, which provides a bridge of electrocortical signals at the mesoscopic and macroscopic levels, thereby facilitating the investigation of nociceptive information processing in the brain.

We developed a technique that simultaneously records both electrocorticography and local field potentials in response to nociceptive laser stimuli from freely moving rats. This technique helps establish a direct relationship of electrocortical signals at the mesoscopic and macroscopic levels, which facilitates the investigation of nociceptive information processing in the brain.

在自由运动大鼠的反应中, 皮质局部电位和电皮质图谱的同时记录

Electrocortical responses, elicited by laser heat pulses that selectively activate nociceptive free nerve endings, are widely used in many animal and ...

Induction of an Isoelectric Brain State to Investigate the Impact of Endogenous Synaptic Activity on Neuronal Excitability In Vivo

The way neurons process information depends both on their intrinsic membrane properties and on the dynamics of the afferent synaptic network. In particular, endogenously-generated network activity, which strongly varies as a function of the state of vigilance, significantly modulates neuronal computation. To investigate how different spontaneous cerebral dynamics impact single neurons&#39; integrative properties, we developed a new experimental strategy in the rat consisting in suppressing in vivo all cerebral activity by means of a systemic injection of a high dose of sodium pentobarbital. Cortical activities, continuously monitored by combined electrocorticogram (ECoG) and intracellular recordings are progressively slowed down, leading to a steady isoelectric profile. This extreme brain state, putting the rat into a deep comatose, was carefully monitored by measuring the physiological constants of the animal throughout the experiments. Intracellular recordings allowed us to characterize and compare the integrative properties of the same neuron embedded into physiologically relevant cortical dynamics, such as those encountered in the sleep-wake cycle, and when the brain was fully silent.

Induction of an Isoelectric Brain State to Investigate the Impact of Endogenous Synaptic Activity on Neuronal Excitability In Vivo

This procedure performs long-lasting in vivo intracellular recordings from single neurons during physiologically relevant cerebral states and after complete abolition of ongoing electrical activities, resulting in an isoelectric brain state. The physiological constants of the animal are carefully monitored during the transition to the artificial comatose condition.

等电脑国家的诱导研究内源性突触活动对神经元兴奋性影响<em&gt;在体内</em

The way neurons process information depends both on their intrinsic membrane properties and on the dynamics of the afferent synaptic network. In ...

神经科学

Extracellular Recording of Neuronal Activity Combined with Microiontophoretic Application of Neuroactive Substances in Awake Mice

Differences in the activity of neurotransmitters and neuromodulators, and consequently different neural responses, can be found between anesthetized and awake animals. Therefore, methods allowing the manipulation of synaptic systems in awake animals are required in order to determine the contribution of synaptic inputs to neuronal processing unaffected by anesthetics. Here, we present methodology for the construction of electrodes to simultaneously record extracellular neural activity and release multiple neuroactive substances at the vicinity of the recording sites in awake mice. By combining these procedures, we performed microiontophoretic injections of gabazine to selectively block GABAA receptors in neurons of the inferior colliculus of head-restrained mice. Gabazine successfully modified neural response properties such as the frequency response area and stimulus-specific adaptation. Thus, we demonstrate that our methods are suitable for recording single-unit activity and for dissecting the role of specific neurotransmitter receptors in auditory processing.
The main limitation of the described procedure is the relatively short recording time (~3 hr), which is determined by the level of habituation of the animal to the recording sessions. On the other hand, multiple recording sessions can be performed in the same animal. The advantage of this technique over other experimental procedures used to manipulate the level of neurotransmission or neuromodulation (such as systemic injections or the use of optogenetic models), is that the drug effect is confined to the local synaptic inputs to the target neuron. In addition, the custom-manufacture of electrodes allows adjustment of specific parameters according to the neural structure and type of neuron of interest (such as the tip resistance for improving the signal-to-noise ratio of the recordings).

We present methods for the construction of electrodes to simultaneously record extracellular neural activity and release multiple neuroactive substances at the vicinity of the recording sites in awake mice. This technique allows the detailed analysis of putative local synaptic inputs to the neuron of interest.

神经元电活动外记录与神经活性物质在清醒小鼠微电泳应用相结合

Differences in the activity of neurotransmitters and neuromodulators, and consequently different neural responses, can be found between anesthetized and ...

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 ...

Construction of an Improved Multi-Tetrode Hyperdrive for Large-Scale Neural Recording in Behaving Rats

Monitoring the activity patterns of a large population of neurons over many days in awake animals is a valuable technique in the field of systems neuroscience. One key component of this technique consists of the precise placement of multiple electrodes into desired brain regions and the maintenance of their stability. Here, we describe a protocol for the construction of a 3D-printable hyperdrive, which includes eighteen independently adjustable tetrodes, and is specifically designed for in vivo extracellular neural recording in freely behaving rats. The tetrodes attached to the microdrives can either be individually advanced into multiple brain regions along the track, or can be used to place an array of electrodes into a smaller area. The multiple tetrodes allow for simultaneous examination of action potentials from dozens of individual neurons, as well as local field potentials from populations of neurons in the brain during active behavior. In addition, the design provides for simpler 3D drafting software that can easily be modified for differing experimental needs.

We present the construction of a 3D-printable hyperdrive with eighteen independently adjustable tetrodes. The hyperdrive is designed to record brain activity in freely behaving rats over a period of several weeks.

一种改进的多电子四极超光速引擎在行为大鼠大尺度神经记录中的构建

Monitoring the activity patterns of a large population of neurons over many days in awake animals is a valuable technique in the field of systems ...

Recording and Modulation of Epileptiform Activity in Rodent Brain Slices Coupled to Microelectrode Arrays

Temporal lobe epilepsy (TLE) is the most common partial complex epileptic syndrome and the least responsive to medications. Deep brain stimulation (DBS) is a promising approach when pharmacological treatment fails or neurosurgery is not recommended. Acute brain slices coupled to microelectrode arrays (MEAs) represent a valuable tool to study neuronal network interactions and their modulation by electrical stimulation. As compared to conventional extracellular recording techniques, they provide the added advantages of a greater number of observation points and a known inter-electrode distance, which allow studying the propagation path and speed of electrophysiological signals. However, tissue oxygenation may be greatly impaired during MEA recording, requiring a high perfusion rate, which comes at the cost of decreased signal-to-noise ratio and higher oscillations in the experimental temperature. Electrical stimulation further stresses the brain tissue, making it difficult to pursue prolonged recording/stimulation epochs. Moreover, electrical modulation of brain slice activity needs to target specific structures/pathways within the brain slice, requiring that electrode mapping be easily and quickly performed live during the experiment. Here, we illustrate how to perform the recording and electrical modulation of 4-aminopyridine (4AP)-induced epileptiform activity in rodent brain slices using planar MEAs. We show that the brain tissue obtained from mice outperforms rat brain tissue and is thus better suited for MEA experiments. This protocol guarantees the generation and maintenance of a stable epileptiform pattern that faithfully reproduces the electrophysiological features observed with conventional field potential recording, persists for several hours, and outlasts sustained electrical stimulation for prolonged epochs. Tissue viability throughout the experiment is achieved thanks to the use of a small-volume custom recording chamber allowing for laminar flow and quick solution exchange even at low (1 mL/min) perfusion rates. Quick MEA mapping for real-time monitoring and selection of stimulating electrodes is performed by a custom graphic user interface (GUI).

We illustrate how to perform recording and electrical modulation of 4-aminopyridine-induced epileptiform activity in rodent brain slices using microelectrode arrays. A custom recording chamber maintains tissue viability throughout prolonged experimental sessions. Live electrode mapping and selection of stimulating pairs are performed by a custom graphical user interface.

鼠脑切片偶联微电极阵列癫痫活性的记录与调控

Temporal lobe epilepsy (TLE) is the most common partial complex epileptic syndrome and the least responsive to medications. Deep brain stimulation (DBS) ...

<meta charset="utf8"></head><body>小鼠深部脑干结构的倾斜硅多电极记录

Acute Single-Unit Multi-Electrode Recordings from the Brainstem of Head-Fixed Mice

Silicon multielectrode-based recordings are increasingly popular for studying neuronal activity at the temporal resolution of action potentials in many brain regions. However, recording neuronal activity from deep caudal structures like the brainstem using multi-channel probes remains challenging. A significant concern is finding a trajectory for probe insertion that avoids large blood vessels, such as the superior sagittal venous sinus and the transverse venous sinus. Injuring these large veins can cause extensive bleeding, damage to the underlying brain tissue, and potentially death. This approach describes targeting brainstem structures by coupling anterior coordinates with an angled approach, allowing the recording probe to penetrate the brain below high-risk vascular structures. Compared to a strictly vertical approach, the angled approach maximizes the number of brain regions that can be targeted. Using this strategy, the ventrolateral periaqueductal gray (vlPAG), a brainstem region associated with REM sleep, can be reproducibly and reliably accessed to obtain single-unit, multi-electrode recordings in head-fixed mice before and during sevoflurane anesthesia. The ability to record neuronal activity in the vlPAG and surrounding nuclei with high temporal resolution is a step forward in advancing the understanding of the relationship between REM sleep and anesthesia.

<meta charset="utf8"></head><body>作者聚焦：研究小鼠麻醉诱导的睡眠通路和神经元兴奋性

This protocol describes a method to obtain in vivo, high-density single-neuron recordings from the brainstem of head-fixed mice. This approach is deployed to measure the action potential firing of neurons in the ventrolateral periaqueductal gray - a brainstem region inactive during Rapid Eye Movement (REM) sleep - before and during general anesthesia.

来自头部固定小鼠脑干的急性单单位多电极记录

Silicon multielectrode-based recordings are increasingly popular for studying neuronal activity at the temporal resolution of action potentials in many ...

急性

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此视频中的章节

用于体内的无线双向接口记录和刺激自由行为大鼠的神经活动

在自由运动大鼠的反应中, 皮质局部电位和电皮质图谱的同时记录

等电脑国家的诱导研究内源性突触活动对神经元兴奋性影响

神经元电活动外记录与神经活性物质在清醒小鼠微电泳应用相结合

啮齿动物联合局部脑电图和局部电位的同步记录

一种改进的多电子四极超光速引擎在行为大鼠大尺度神经记录中的构建

鼠脑切片偶联微电极阵列癫痫活性的记录与调控

来自头部固定小鼠脑干的急性单单位多电极记录

来自头部固定小鼠脑干的急性单单位多电极记录

来自头部固定小鼠脑干的急性单单位多电极记录