Name: automatic detection of highly organized theta oscillations in the murine eeg
Uploaded: 2017-03-10T13:00:00.000+00:00
Duration: 9 min 35 s
Description: Watch this Scientific Journal Video about Automatic Detection of Highly Organized Theta Oscillations in the Murine EEG at JoVE.com

The authors would like to thank Dr. Christina Ginkel (German Center for Neurodegenerative Diseases, DZNE) and Dr. Robert Stark (DZNE) for their assistance with animal breeding and animal healthcare. This work was financially supported by the Federal Institute for Drugs and Medical Devices (Bundesinstitut f&#252;r Arzneimittel und Medizinprodukte, BfArM), Bonn, Germany.

所有的动物实验是根据当地和机构理事会关于动物保健（波恩大学，BfArM，LANUV，德国）的指导方针进行。此外，所有的动物实验是按照上级的立法， 如 1986年11月24日（86/609 / EEC）的欧洲共同体理事会指令，或个别区域或国家的法律进行。具体的努力是为了最大限度地减少使用的动物的数量，以及它们的痛苦。 1.动物住房和脑电图记录条件<ol><li>家鼠过滤器顶笼或使用独立通风笼。 </li><li>从动物设施在特殊的实验室室，适合植入动物和遥测记录传输小鼠通风机箱。 </li><li>执行所有动物实验，包括脑电图电极的植入和随后的录音，标准条件（22℃透射电镜023，12小时/ 12小时明-暗循环，50％-60％的相对湿度，噪声衰减等 ）18。 </li><li>之前射频发射机注入，房子的动物中的3组- 4清楚II型聚碳酸酯笼子，食物和水随意 。不要分离单个老鼠，因为这可能导致紧张和随后的实验和结果产生干扰。 </li><li>不要使用开放式的住房条件，但实验和录制过程中，而使用通风机箱。 </li></ol> 2.无线电遥测脑电图植入电极和脑电图记录<ol><li> 麻醉用注射麻醉剂， 例如，ketaminehydrochloride / xylazinehydrochloride（100/10毫克/千克，腹膜内，IP）或吸入麻醉剂， 如异氟烷17,18小鼠。 <ol><li>对于异氟烷麻醉，将鼠标定位到一个感应Çhamber用4-5％异氟醚和0.8-1％的氧气或卡波金（5％CO 2和95％O 2）。 </li><li>放置一个硅面罩上的动物，以控制麻醉的期望深度，并使用清除系统避免实验者暴露于异氟醚的鼻/口。 </li><li>使用注射麻醉剂， 如 esketaminhydrochloride（100毫克/千克，IP）和xylazinehydrochloride（10毫克/千克，IP），如果吸入麻醉不可用。 </li><li>通过检查尾巴捏反射，脚捏反射和呼吸速率监测麻醉深度。注意， 经由气管插管进行人工呼吸不在小鼠中必要的。 </li></ol></li><li> 植入射频发射机插入动物背部皮下袋。 <ol><li>从头皮去除体毛和具有两个消毒剂， 即 70％的乙醇和碘基擦洗预处理剃头皮。 </li><li>使用手术刀，使从前额到nucheal区域头皮中线切开。 </li><li>从颈部切口开始，通过使用外科剪刀或外科手术探针进行钝器解剖准备在动物背部的一侧的皮下袋中。 </li><li>将发射器放入皮下囊和灵活的发射器通向袋，以及存放多余的长度。要特别注意防止手术部位和发射器植入物的污染。正确地隔离使用窗帘非无菌区无菌。 </li></ol></li><li>将实验动物的立体框架， 例如，一个计算机化的3D立体设备。修正了使用鼻夹和耳棒头骨。 </li><li>预处理用0.3％ 的 H 2 O 2的头骨从头骨移除进一步的组织并照亮颅缝和craniometrics地标，前囟和lambda。 </li><li>在选择的坐标钻孔在以最大速度的无压模式下使用一个高速神经外科钻（参见步骤2.6）。 注:无压力钻井避免钻头和损坏皮质的突然突破。对于开颅手术，建议神经外科高速电钻。选择的0.3标准钻头直径 - 0.5毫米，这取决于电极的直径。 </li><li>慎重选择电极型，考虑到阻抗，直径，涂料等。 注:聚对二甲苯被覆钨或不锈钢电极是最常见的。电极类型应根据实验要求来选择。作为一个先发制人的手法，用消毒70％乙醇植入前电极头。注意，电极涂覆不允许用于加热消毒。 </li><li>对于海马CA1脑电图记录，在以下坐标立体定向钻孔:前囟门，-2毫米;中侧，1.5毫米（右半球）;背腹，1.3毫米（圆盾ŧ地区:大角ammonis（CA1）锥体细胞层）。为参考电极，在以下立体坐标钻小脑皮层以上的孔:前囟，-6.2毫米，中侧0毫米;背腹0毫米。 注:小脑电极用作pseudoreference电极，如小脑是一个相当无声大脑区域。立体定位坐标被从标准鼠标脑图谱而得。 </li><li>到电极的插入之前，将它们缩短到所需长度。机械地夹在电极到发射器的不锈钢螺旋的颅外一部分。 注:焊接应尽量避免，因为这会在系统中引入大量噪声。 </li><li>该电极连接到立体定位装置的垂直臂和根据上述立体坐标插入电极。 </li><li>采用固定玻璃离子水门汀电极，等到水泥已经完全硬化。 </li><li>关闭头皮使用过和悬停缝线用非可吸收的5-0或6-0缝合材料。 </li><li>对于术后疼痛管理，管理卡洛芬（5毫克/千克，皮下，SC），每日一次，连续4天，植入后。需要注意的是卡布洛芬应在首次切口（步骤2.2.2）注入。 </li><li>让动物10恢复 - 录音和/或注塑实验开始前14天植入后。 </li></ol> 3.西塔振荡与药理诱导的自发录音<ol><li>激活使用磁性开关的射频发射机。放置动物与接收器上的板其家笼。至少24-48小时进行长期海马脑电图记录。 注:脑电图振幅和长期记录脑电图频率特性的分析提供了详细的洞察成θ振荡的昼夜依赖性以及它们与特定行为和关联认知条件/任务。始终结合脑电图记录与实验动物的视频监控。 </li><li>为THETA振荡药理感应，辖氨基甲酸酯（800毫克/千克IP）或毒蕈碱受体激动剂的单一剂量，例如毛果芸香碱（10毫克/千克IP），槟榔碱（0.3毫克/千克IP）或氧化震颤素的单剂量（0.03毫克/千克IP）。预处理与N-甲基东（0.5毫克/千克IP）的小鼠，以避免外周毒蕈碱反应。新鲜溶解在0.9％NaCl或林格氏液所有药物。 注:更高的毒蕈碱受体激动剂的剂量可能导致癫痫发作感应的实验动物。同时认为，这里给出的剂量表示需要在受调查的鼠标线前剂量 - 效应研究的里程碑。注意，聚氨酯是需要在储存和处理适当的预防措施诱变剂和致癌物质。 </li><li>注射阿托品（50毫克/千克，IP）来区分阿托品敏感型II从阿托品不敏感型我THETA振荡。 注:阿托品用量为一次物种和应变依赖和需要事先剂量 - 效应评价。阿托品注射液的最佳时间点取决于毒蕈碱受体激动剂的药效学。 II型theta的识别，阿托品注射氨基甲酸酯给药后1小时，建议。 </li><li>尽量避免的几种药物的后续申请，如全身性药物施用改变全局转录和翻译模式，这影响后续脑电图记录。需要注意的是短暂的振荡THETA也可以通过感官刺激诱发，如tail-或爪子捏。 </li><li>萃取物/出口代表脑电图数据集的前阶段（基线），并从总的脑电图记录作为ASCI或TXT文件后喷射阶段的，考虑到所施加的药物的药代动力学和个体研究协议的要求。 </li></ol> 4。脑电图电极放置的验证<ol><li>通过将它们放置在一个孵育室安乐死的动物和引入100％的二氧化碳。使用的每与加入到孵育室的现有空气二氧化碳分腔室容积的10-30％的填充率;这将导致以最小的应力给动物迅速失去知觉。 </li><li>从室中取出鼠标一旦出现呼吸停止，暗淡的眼睛颜色持续2-3分钟。 </li><li>切不锈钢电极和外植体射频发射器。用剪刀或者断头台斩首鼠标轻轻操纵手术剪刀和镊子取出从脑颅大脑。 </li><li>固定在4％低聚甲醛的大脑在磷酸盐缓冲盐水（PBS）（pH 7.4）中过夜。对于冷冻保护，在4℃的大脑转移到30％的葡萄糖，并将其储存，直到进一步处理。 注意:多聚甲醛是由2012 OSHA哈兹认为是有害的ARD通信标准（29 CFR 1910.1200）。采取必要的预防措施:使用个人防护用品，保证适当的通风，避免形成粉尘。此外，除去火源，并采取防止静电放电的措施。低聚甲醛不应该被释放到环境中。 </li><li>安装在使用粘接剂的低温恒温器的组织保持器所提取的脑切大脑成40 - 75微米的冠状切片。安装片到玻片上，并使用标准的组织学方法与尼氏蓝染色他们。这个过程将可视化反映的是前电极位置的分支通道。请注意，它也有可能使用vibroslicer从天然脑切冠状切片</li><li>只合并那些符合正确的脑电图电极放置标准的动物;对于CA1区，深电极的尖端应该CA1区锥体细胞层内进行本地化。 </li></ol><ol><li>记录CA1海马脑电图与没有先验滤波器截止适当的采样速率。 注:采样速率，这是发射器 - 特异性的，确定脑电图分析的频率上限。 </li><li>处理与分析软件的记录的数据。程序的时间-频率分析，并与用于分析方法的适当的控制（ 图5）20定做程序计算。 </li><li>切断记录脑电图段用各1小时的长度。使用快速计算机处理器，因为计算时间是很高的。此外，利用软件，可以在多个内核并行20计算。 </li></ol> 6.脑电图数据分析<ol><li>分析使用复杂Morlet小波来计算的频率和振荡的振幅数据段。 注:此小波（ 例如，Ψ（X）=（πB）（ - 1/2）EXP（2 Iπçx）的EXP（-x 2 / b）中 ，其中b是带宽参数，c中的中心频率，和我虚数单位）经常在文献中研究脑电图被应用数据，因为它保证了在频率和时间两者23,24最佳分辨率。 </li><li>使用带宽参数和中心频率设置，尤其是权重，同时还不能忽略足够的时间分辨率，频率分辨率，以区分在0.1赫兹级频率的差异。 注:伽玛带神经过程是短暂的25，这也可以容纳如此THETA节奏。因此，分析方法必须考虑足够的时间分辨率。 </li><li>在0.2的频率范围分析的EEG数据 - 12赫兹，0.1 Hz的步长，因此包括典型的增量，该TA，和α的频率范围。 </li><li>设立的θ频率架构的论述，这可以代替THETA振荡的标准目视检查的自动化，复杂的分析用具;此过程被称为的θ的检测方法（TDM）的。 </li><li>计算中的θ频率范围（3.5-8.5赫兹）的最大振幅的商并且在上三角频率范围为每2.5秒的时间窗的最大振幅（2-3.4赫兹）。 注:此商数的值用作度量来决定，如果发生了theta电振荡。的θ频率范围的定义可以根据不同的功能状态和神经解剖学电路/系统上进行分析而变化。 </li><li>分类段为"THETA振荡时期，"如果这部分期间计算出的比例是1.5以上。 注意:这保证了最大THETA振幅相关2.5 - S的脑电图期间比在上部三角形带的幅度高至少50％的分割。注意，该比率可能取决于所用的线路和/或物种需要适应。的2.5秒的时间间隔表示为一个theta电振荡的最小持续时间，防止某些嘈杂历元的假阳性检测，并位于其他出版物19，26的范围定义内。上部三角形的频率范围作为控制频带因为慢波睡眠，这是THETA活动期间高度衰减期间在非-θ时期出现生理学相关的增量活性，例如，。 </li><li>重复此过程，每1小时部分;因此，每一个部分包括与每2.5秒长度1,440脑电图段。 </li><li>统计学评估所识别THETA振荡历元中的数据。 </li><li>计算检测THETA历元的总持续时间的统计信息;不同的或预定义的组;循环，如光/暗;和其他参数。 注:为staticSTICS可以包括t-检验，ANOVA，或MANOVA，取决于变量，组的数量，条件等 </li><li>计算所检测THETA历元的幅度的统计数据，但是只在的θ的频率范围（3.5 - 8.5赫兹）。 </li><li>评估所检测THETA历元的频率的统计数据，但是只在的θ的频率范围（3.5 - 8.5赫兹）。 注意:一THETA划时代的THETA频率被定义为属于一个THETA划时代的最大THETA振幅的频率。 </li></ol>

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The authors have nothing to disclose.

西塔活动是全身神经生理学中心的相关性。它可以在各种脑区域可以观察到，特别是在海马，在那里它与特定行为和认知状态。此外，海马THETA可以药理分化成阿托品敏感的II型和阿托品不敏感型我THETA。 I型被认为是涉及到运动，如走路或跑步27，28，29，30，31，而II型可在警...

睡在大脑中的Theta活动涉及到不同的认知和功能状态，包括觉醒，注意，随意运动，探索行为，注意力的行为，学习和记忆，体感集成和快速眼动（REM）1，2。原则上，可以在不同的脑区中产生THETA活动作为一个有节奏的实体，高度组织化和同步，THETA振荡。下面，我们将集中于THETA活动/被所述septohippocampal系统3,4内产生振荡的分析和定量。内的隔膜，GABA能，谷氨酸，和胆碱能神经元投射到海马和有助于THETA振荡行为的启动和维护。有对海马THETA振荡是否在隔发起的， 即一个持续的讨论， </em&gt;隔起搏器海马跟随模式，（extrahippocampal理论）或本质海马内（海马理论）5，6，7。 不论其来源，海马THETA振荡已经在感兴趣的焦点多年来，尤其是在转基因小鼠模型。这些模型允许深脑电图电极的植入和在特定的认知和行为的任务8海马THETA振荡的记录。海马THETA振荡是在自然界中异源。根据THETA振荡的所谓的二元论，可以阿托品敏感的II型theta和阿托品不敏感的I型THETA 9，10，11之间区分。后者通常可以通过毒蕈碱诱导1 / M <sUB&gt; 3受体激动剂， 如槟榔碱，毛果芸香碱和聚氨酯。然而，氨基甲酸乙酯是一种多目标药物，除了毒蕈碱活化，也施加在其它离子通道实体复杂的影响。 II型THETA，所述毒蕈碱性通路包括M 1的活化/ M 3和随后ģQ / 11（Gα）磷脂酶C介导的活化β1/4（PLCβ1/4），三磷酸肌醇（INSP 3） ，diacylglycerole（DAG），钙离子 ，和蛋白激酶C（PKC）。 PLCβ1和PLCβ4的thetagenesis的作用已在敲除研究使用被验证PLCβ1 - / -和PLCβ4 - / -呈现THETA振荡12，13，14的完全丧失或显著衰减的小鼠。附加M 1，M 3和M 5下游目标（CHA毒蕈碱信令级联nnels /电流）包括各种电导，诸如经由电压依赖性K +通道（K 7节 ）M型K +通道（K M）;超极化K +通道（KS AHP）后慢;泄漏K +通道（K 泄漏 ），可能通过TWIK相关酸敏感的K +通道（TASK1 / 3）;阳离子电流（I CAT），可能通过钠泄漏通道（NALCN）;并经由超极化和环核苷酸门控通道（HCN）我小时 。此外，M 2 / M 4乙酰胆碱受体（乙酰胆碱受体）报告，其中内向整流干扰K +通道3.1（K IR 3.1）和内向整流钾通道3.2（K IR 3.2）15。 目前，市售的分析软件允许快速基于FFT的频率分析， 例如，功率分析（P，毫伏2）或功率谱密度（PSD，毫伏2 /赫兹）。电源或θ波频率范围内的功率谱密度（PSD）的分析只给其活动的全球概览。然而，为了得到详细的洞察认知和行为相关的THETA活性，高度组织THETA振荡的分析是强制性的。高度组织THETA振荡的评估是在神经变性和神经精神性疾病的领域重要性。最具实验性的疾病的研究都使用高度复杂的神经外科手术的方法来记录硬膜外表层和深层颅内脑电图转基因小鼠模型进行的。这些技术包括系绳系统16和无线电遥测设置17,18。西塔振荡可以记录为长期的录音条件下的自发和行为相关的THETA振荡。此外，THETA振荡可以RECOrded以下的药理感应而且下列动物暴露于行为或认知任务或感觉刺激，例如尾部夹持。 早期方法来表征THETA振荡被Csicsvari 等人所述。 19。作者设计为短期THETA分析的半自动化的工具 - 即不适合长时间脑电图记录（15 50分钟）。我们的方法中，这里描述的，允许长期脑电图记录&gt; 48小时20进行的分析。 Csicsvari 等。 10还提到了THETA三角洲率，但提供了高度组织THETA振荡的决心没有门槛。三角洲和θ范围定义符合我们的频率范围的定义。因为它是不明确提到，我们假设基于FFT的方法用于由Csicsvari 等。计算的θ-Δ频带的功率。这个再次从我们的方法明显不同，因为我们计算上的大量频率尺度（频率步骤Δ（F）= 0.05赫兹）基于小波的振幅，导致高得多的精度。独具一格的分析脑电图时代的持续时间是类似于我们的定义。 Klausberger 等。 21也借助THETA三角比长期脑电图记录的分析。但是，也有相对于我们的方法三大差别:i）所述脑电图历元持续时间要长得多， 即至少为6秒; ⅱ）的θ-Δ比设定为4，这是比我们阈高得多，并涉及到不同的频率范围定义;和iii）电源定义很可能是基于FFT的方法，它缺乏精度高，特别是对于非常短的时间窗口（2秒， 即，5个循环为2.5赫兹的频率）振荡。在这种情况下，一个基于小波的过程是更可取的。由卡普兰等人的研究。 22只计算THETA功率而忽略了THETA三角洲功率比。因此，卡普兰接近22不能伴随着高或低的增量认知丰富THETA流程进行区分。 在下面的协议，我们将提出我们的分析基于小波变换的方法来可靠地分析海马脑电图记录高度组织THETA振荡小鼠。由于该过程自动工作，它可以应用到大型数据集和长期EEG测量。

<table><tbody><tr><td>Carprofen (Rimadyl VET - Injektionsl&amp;ouml;sung)</td><td>Pfizer</td><td>PZN 0110208208</td><td>20ml</td></tr><tr><td>binocular surgical magnification microscope</td><td>Zeiss Stemi 2000</td><td>0000001003877, 4355400000000, 0000001063306, 4170530000000, 4170959255000, 4551820000000, 4170959040000, 4170959050000</td><td></td></tr><tr><td>Dexpanthenole (Bepanthen Wund- und Heilsalbe)</td><td>Bayer</td><td>PZN: 1578818</td><td></td></tr><tr><td>drapes (sterile)</td><td>Hartmann</td><td>PZN 0366787</td><td></td></tr><tr><td>70% ethanol</td><td>Carl Roth</td><td>9065.5</td><td></td></tr><tr><td>0.3% / 3% hydrogene peroxide solution</td><td>Sigma</td><td>95321</td><td>30% stock solution</td></tr><tr><td>gloves (sterile)</td><td>Unigloves</td><td>1570</td><td></td></tr><tr><td>dental glas ionomer cement</td><td>KentDental /NORDENTA</td><td>957 321</td><td></td></tr><tr><td>heat-based surgical instrument sterilizer</td><td>F.S.T.</td><td>18000-50</td><td></td></tr><tr><td>high-speed dental drill</td><td>Adeor</td><td>SI-1708</td><td></td></tr><tr><td>Inhalation narcotic system (isoflurane)</td><td>Harvard Apparatus GmbH</td><td>34-1352, 10-1340, 34-0422, 34-1041, 34-0401, 34-1067, 72-3044, 34-0426, 34-0387, 34-0415, 69-0230</td><td></td></tr><tr><td>Isoflurane</td><td>Baxter 250 ml</td><td>PZN 6497131</td><td></td></tr><tr><td>Ketamine</td><td>Pfizer</td><td>PZN 07506004</td><td></td></tr><tr><td>Lactated Ringer&#039;s solution (sterile)</td><td>Braun</td><td>L7502</td><td></td></tr><tr><td>Nissl staining solution</td><td>Armin Baack</td><td>BAA31712159</td><td></td></tr><tr><td>pads (sterile)</td><td>ReWa Krankenhausbedarf</td><td>2003/01</td><td></td></tr><tr><td>Steel and tungsten electrodes parylene coated</td><td>FHC Inc., USA</td><td>UEWLGESEANND</td><td></td></tr><tr><td>stereotaxic frame</td><td>Neurostar</td><td>51730M</td><td>ordered at Stoelting</td></tr><tr><td>(Stereo Drive-New Motorized Stereotaxic)</td><td></td><td></td><td></td></tr><tr><td>tapes (sterile)</td><td>BSN medical GmbH &amp;amp; Co. KG</td><td>626225</td><td></td></tr><tr><td>TA10ETA-F20</td><td>DSI</td><td>270-0042-001X</td><td>Radiofrequency transmitter 3.9 g, 1.9 cc, input voltage range &amp;plusmn; 2.5 mV, channel bandwidth (B) 1 - 200 Hz, nominal sampling rate (f) 1,000 Hz (f = 5B) temperature operating range 34 - 41 &amp;deg;C warranted battery life 4 months</td></tr><tr><td>TL11M2-F20EET</td><td>DSI</td><td>270-0124-001X</td><td>Radiofrequency transmitter 3.9 g, 1.9 cc, input voltage range &amp;plusmn; 1.25 mV, channel bandwidth (B) 1 - 50 Hz, nominal sampling rate (f) 250 Hz (f = 5B) temperature operating range 34 - 41 &amp;deg;C warranted battery life 1.5 months</td></tr><tr><td>Vibroslicer 5000 MZ</td><td>Electron Microscopy Sciences</td><td>5000-005</td><td></td></tr><tr><td>Xylazine (Rompun)</td><td>Bayer</td><td>PZN: 1320422</td><td></td></tr><tr><td>Matlab</td><td>Mathworks Inc.</td><td></td><td>programming, computing and visualization software</td></tr><tr><td>SPSS</td><td>IBM</td><td></td><td>statistical analysis software</td></tr></tbody></table>

automatic detection of highly organized theta oscillations in the murine eeg

西塔活性在septohippocampal系统中产生，并且可以使用深海马电极和植入脑电图（EEG）无线电遥测或系绳系统的方法进行记录。药理学上，海马THETA是多相（见二元理论），并且可以分化为I型和II型2θ。这些个别脑电图亚型都与特定的认知和行为状态，如觉醒，勘探，学习和记忆，更高的综合功能，等等。在神经变性疾病如阿尔茨海默氏症，结构和septohippocampal系统的功能的改变可导致受损THETA活性/振荡。海马脑电图的标准定量分析包括快速傅里叶变换（FFT）为基础的时频分析。但是，此过程不提供有关特别是在一般和高度组织THETA振荡THETA活动的详细信息。为了获得DETA在海马高度组织THETA振荡ILED的信息，我们已经开发了一种新的分析方法。该方法允许高度组织THETA振荡和其频率特性的持续时间的时间和成本有效的量化。

西塔活性可以被记录在一个大范围的中枢神经系统（CNS）区域中。在这里，我们提出从小鼠海马THETA振荡的分析。在不同的行为和认知状态，可能会发生这种振荡。强烈建议来分析自发的长期任务相关的短期和药理学诱导条件下THETA振荡。 图1显示了控制条件下，具有代表性的海马CA1记录。如果动物是不是在一个自发的"THETA国家"海马脑电图的?...

Watch this Scientific Journal Video about Automatic Detection of Highly Organized Theta Oscillations in the Murine EEG at JoVE.com

Automatic Detection of Highly Organized Theta Oscillations in the Murine EEG

Theta activity in the hippocampus is related to specific cognitive and behavioral stages. Here, we describe an analytical method to detect highly-organized theta oscillations within the hippocampus using a time-frequency (i.e., wavelet analysis)-based approach.

高度组织的Theta振荡在鼠脑的自动检测

Theta activity is generated in the septohippocampal system and can be recorded using deep intrahippocampal electrodes and implantable ...

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9.1K 次观看.  University of Cologne. Theta activity in the hippocampus is related to specific cognitive and behavioral stages. Here, we describe an analytical method to detect highly-organized theta oscillations within the hippocampus using a time-frequency (i.e., wavelet analysis)-based approach.

视频: Automatic Detection of Highly Organized Theta Oscillations in the Murine EEG

EEG Mu Rhythm in Typical and Atypical Development

Electroencephalography (EEG) is an effective, efficient, and noninvasive method of assessing and recording brain activity. Given the excellent temporal resolution, EEG can be used to examine the neural response related to specific behaviors, states, or external stimuli. An example of this utility is the assessment of the mirror neuron system (MNS) in humans through the examination of the EEG mu rhythm. The EEG mu rhythm, oscillatory activity in the 8-12 Hz frequency range recorded from centrally located electrodes, is suppressed when an individual executes, or simply observes, goal directed actions. As such, it has been proposed to reflect activity of the MNS. It has been theorized that dysfunction in the mirror neuron system (MNS) plays a contributing role in the social deficits of autism spectrum disorder (ASD). The MNS can then be noninvasively examined in clinical populations by using EEG mu rhythm attenuation as an index for its activity. The described protocol provides an avenue to examine social cognitive functions theoretically linked to the MNS in individuals with typical and atypical development, such as ASD.&#160;

Assessment of the EEG mu rhythm provides a unique methodology for examining brain activity and when combined with behaviorally based assays, can be a powerful tool for elucidating aspects of social cognition, such as imitation, in clinical populations.

脑电图沐节奏典型和非典型发展

Electroencephalography (EEG) is an effective, efficient, and noninvasive method of assessing and recording brain activity. Given the excellent temporal ...

科研

医学

Long-term Continuous EEG Monitoring in Small Rodent Models of Human Disease Using the Epoch Wireless Transmitter System

Many progressive neurologic diseases in humans, such as epilepsy, require pre-clinical animal models that slowly develop the disease in order to test interventions at various stages of the disease process. These animal models are particularly difficult to implement in immature rodents, a classic model organism for laboratory study of these disorders. Recording continuous EEG in young animal models of seizures and other neurological disorders presents a technical challenge due to the small physical size of young rodents and their dependence on the dam prior to weaning. Therefore, there is not only a clear need for improving pre-clinical research that will better identify those therapies suitable for translation to the clinic but also a need for new devices capable of recording continuous EEG in immature rodents. Here, we describe the technology behind and demonstrate the use of a novel miniature telemetry system, specifically engineered for use in immature rats or mice, which is also effective for use in adult animals.

Here we demonstrate the use of a wireless enabling technology for electroencephalogram (EEG) in neonatal rodent models of human disease. With telemetry, there are no encumbering connections, thus allowing natural behaviors.

在人类疾病中使用大纪元无线传输系统的小型啮齿动物模型的长期连续监测脑电图

Many progressive neurologic diseases in humans, such as epilepsy, require pre-clinical animal models that slowly develop the disease in order to test ...

神经科学

Non-restraining EEG Radiotelemetry: Epidural and Deep Intracerebral Stereotaxic EEG Electrode Placement

Implantable EEG radiotelemetry is of central relevance in the neurological characterization of transgenic mouse models of neuropsychiatric and neurodegenerative diseases as well as epilepsies. This powerful technique does not only provide valuable insights into the underlying pathophysiological mechanisms, i.e., the etiopathogenesis of CNS related diseases, it also facilitates the development of new translational, i.e., therapeutic approaches. Whereas competing techniques that make use of recorder systems used in jackets or tethered systems suffer from their unphysiological restraining to semi-restraining character, radiotelemetric EEG recordings overcome these disadvantages. Technically, implantable EEG radiotelemetry allows for precise and highly sensitive measurement of epidural and deep, intracerebral EEGs under various physiological and pathophysiological conditions. First, we present a detailed protocol of a straight forward, successful, quick and efficient technique for epidural (surface) EEG recordings resulting in high-quality electrocorticograms. Second, we demonstrate how to implant deep, intracerebral EEG electrodes, e.g., in the hippocampus (electrohippocampogram). For both approaches, a computerized 3D stereotaxic electrode implantation system is used. The radiofrequency transmitter itself is implanted into a subcutaneous pouch in both mice and rats. Special attention also has to be paid to pre-, peri- and postoperative treatment of the experimental animals. Preoperative preparation of mice and rats, suitable anesthesia as well as postoperative treatment and pain management are described in detail.

Non-restraining EEG radiotelemetry is a valuable methodological approach to record in vivo long-term electroencephalograms from freely moving rodents. This detailed protocol describes stereotaxic epidural and deep intracerebral electrode placement in different brain regions in order to obtain reliable recordings of CNS rhythmicity and CNS related behavioral stages.

非约束脑电遥测:硬膜外和脑深部脑电立体定位放置电极

Implantable EEG radiotelemetry is of central relevance in the neurological characterization of transgenic mouse models of neuropsychiatric and ...

Tuning in the Hippocampal Theta Band In Vitro: Methodologies for Recording from the Isolated Rodent Septohippocampal Circuit

This protocol outlines the procedures for preparing and recording from the isolated whole hippocampus, of WT and transgenic mice, along with recent improvements in methodologies and applications for the study of theta oscillations. A simple characterization of the isolated hippocampal preparation is presented whereby the relationship between internal hippocampal theta oscillators is examined together with the activity of pyramidal cells, and GABAergic interneurons, of the cornu ammonis-1 (CA1) and subiculum (SUB) areas. Overall, we show that the isolated hippocampus is capable of generating intrinsic theta oscillations in vitro and that rhythmicity generated within the hippocampus can be precisely manipulated by optogenetic stimulation of parvalbumin-positive (PV) interneurons. The in vitro isolated hippocampal preparation offers a unique opportunity to use simultaneous field and intracellular patch-clamp recordings from visually-identified neurons to better understand the mechanisms underlying theta rhythm generation.

Tuning in the Hippocampal Theta Band In Vitro: Methodologies for Recording from the Isolated Rodent Septohippocampal Circuit

Here, we present a protocol for recording rhythmic neuronal network theta and gamma oscillations from an isolated whole hippocampal preparation. We describe the experimental steps from extraction of the hippocampus to details of field, unitary and whole-cell patch clamp recordings as well as optogenetic pacing of the theta rhythm.

调整在海马Theta乐队<em&gt;体外</em&gt;:从孤立的啮齿动物皮下注射电路记录的方法

This protocol outlines the procedures for preparing and recording from the isolated whole hippocampus, of WT and transgenic mice, along with recent ...

Interictal High Frequency Oscillations Detected with Simultaneous Magnetoencephalography and Electroencephalography as Biomarker of Pediatric Epilepsy

Crucial to the success of epilepsy surgery is the availability of a robust biomarker that identifies the Epileptogenic Zone (EZ). High Frequency Oscillations (HFOs) have emerged as potential presurgical biomarkers for the identification of the EZ in addition to Interictal Epileptiform Discharges (IEDs) and ictal activity. Although they are promising to localize the EZ, they are not yet suited for the diagnosis or monitoring of epilepsy in clinical practice. Primary barriers remain: the lack of a formal and global definition for HFOs; the consequent heterogeneity of methodological approaches used for their study; and the practical difficulties to detect and localize them noninvasively from scalp recordings. Here, we present a methodology for the recording, detection, and localization of interictal HFOs from pediatric patients with refractory epilepsy. We report representative data of HFOs detected noninvasively from interictal scalp EEG and MEG from two children undergoing surgery.
The underlying generators of HFOs were localized by solving the inverse problem and their localization was compared to the Seizure Onset Zone (SOZ) as this was defined by the epileptologists. For both patients, Interictal Epileptogenic Discharges (IEDs) and HFOs were localized with source imaging at concordant locations. For one patient, intracranial EEG (iEEG) data were also available. For this patient, we found that the HFOs localization was concordant between noninvasive and invasive methods. The comparison of iEEG with the results from scalp recordings served to validate these findings. To our best knowledge, this is the first study that presents the source localization of scalp HFOs from simultaneous EEG and MEG recordings comparing the results with invasive recordings. These findings suggest that HFOs can be reliably detected and localized noninvasively with scalp EEG and MEG. We conclude that the noninvasive localization of interictal HFOs could significantly improve the presurgical evaluation for pediatric patients with epilepsy.

High Frequency Oscillations (HFOs) have emerged as presurgical biomarkers for the identification of the epileptogenic zone in pediatric patients with medically refractory epilepsy. A methodology for the noninvasive recording, detection, and localization of HFOs with simultaneous scalp electroencephalography (EEG) and magnetoencephalography (MEG) is presented.

同时用脑磁图和脑电图为小儿癫痫的生物标志物检测发作间期高频振动

Crucial to the success of epilepsy surgery is the availability of a robust biomarker that identifies the Epileptogenic Zone (EZ). High Frequency ...

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 ...

Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice

Extensive data on relationships of neural network oscillations to behavior and organization of neuronal discharge across brain regions call for new tools to selectively manipulate brain rhythms. Here we describe an approach combining projection-specific optogenetics with extracellular electrophysiology for high-fidelity control of hippocampal theta oscillations (5-10 Hz) in behaving mice. The specificity of the optogenetic entrainment is achieved by targeting channelrhodopsin-2 (ChR2) to the GABAergic population of medial septal cells, crucially involved in the generation of hippocampal theta oscillations, and a local synchronized activation of a subset of inhibitory septal afferents in the hippocampus. The efficacy of the optogenetic rhythm control is verified by a simultaneous monitoring of the local field potential (LFP) across lamina of the CA1 area and/or of neuronal discharge. Using this readily implementable preparation we show efficacy of various optogenetic stimulation protocols for induction of theta oscillations and for the manipulation of their frequency and regularity. Finally, a combination of the theta rhythm control with projection-specific inhibition addresses the readout of particular aspects of the hippocampal synchronization by efferent regions.

We describe the use of optogenetics and electrophysiological recordings for selective manipulations of hippocampal theta oscillations (5-10 Hz) in behaving mice. The efficacy of the rhythm entrainment is monitored using local field potential. A combination of opto- and pharmacogenetic inhibition addresses the efferent readout of hippocampal synchronization.

Optogenetic 在行为小鼠海马θ振荡中的作用

Extensive data on relationships of neural network oscillations to behavior and organization of neuronal discharge across brain regions call for new tools ...

Recording Spatially Restricted Oscillations in the Hippocampus of Behaving Mice

The local field potential (LFP) emerges from ion movements across neural membranes. Since the voltage recorded by LFP electrodes reflects the summed electrical field of a large volume of brain tissue, extracting information about local activity is challenging. Studying neuronal microcircuits, however, requires a reliable distinction between truly local events and volume-conducted signals originating in distant brain areas. Current source density (CSD) analysis offers a solution for this problem by providing information about current sinks and sources in the vicinity of the electrodes. In brain areas with laminar cytoarchitecture such as the hippocampus, one-dimensional CSD can be obtained by estimating the second spatial derivative of the LFP. Here, we describe a method to record multilaminar LFPs using linear silicon probes implanted into the dorsal hippocampus. CSD traces are computed along individual shanks of the probe. This protocol thus describes a procedure to resolve spatially restricted neuronal network oscillations in the hippocampus of freely moving mice.

This protocol describes the recording of local field potentials with multi-shank linear silicon probes. Conversion of the signals using current source density analysis allows the reconstruction of local electrical activity in the mouse hippocampus. With this technique, spatially restricted brain oscillations can be studied in freely moving mice.

行为小鼠海马空间受限振荡的记录

The local field potential (LFP) emerges from ion movements across neural membranes. Since the voltage recorded by LFP electrodes reflects the summed ...

Multi-system Monitoring for Identification of Seizures, Arrhythmias and Apnea in Conscious Restrained Rabbits

Patients with ion channelopathies are at a high risk of developing seizures and fatal cardiac arrhythmias. There is a higher prevalence of heart disease and arrhythmias in people with epilepsy (i.e., epileptic heart.) Additionally, cardiac and autonomic disturbances have been reported surrounding seizures. 1:1,000 epilepsy patients/year die of sudden unexpected death in epilepsy (SUDEP). The mechanisms for SUDEP remain incompletely understood. Electroencephalograms (EEG) and electrocardiograms (ECG) are two techniques routinely used in the clinical setting to detect and study the substrates/triggers for seizures and arrhythmias. While many studies and descriptions of this methodology are in rodents, their cardiac electrical activity differs significantly from humans. This article provides a description of a non-invasive method for recording simultaneous video-EEG-ECG-oximetry-capnography in conscious rabbits. As cardiac electrical function is similar in rabbits and humans, rabbits provide an excellent model of translational diagnostic and therapeutic studies. In addition to outlining the methodology for data acquisition, we discuss the analytical approaches for examining neuro-cardiac electrical function and pathology in rabbits. This includes arrhythmia detection, spectral analysis of EEG and a seizure scale developed for restrained rabbits.

Using simultaneous video-EEG-ECG-oximetry-capnography, we developed a methodology to evaluate the susceptibility of rabbit models to develop provoked arrhythmias and seizures. This novel recording system establishes a platform to test the efficacy and safety of therapeutics and can capture the complex cascade of multi-system events that culminate in sudden death.

意识约束兔子中癫痫发作、心律失常和呼吸暂停的多系统监测

Patients with ion channelopathies are at a high risk of developing seizures and fatal cardiac arrhythmias. There is a higher prevalence of heart disease ...

Selective Manipulation of Theta Oscillations in a Mouse Model

Source: Bender, F. et al., <a href="https://app.jove.com/v/57349">Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice.</a> J. Vis. Exp (2018)
This video demonstrates a protocol to modulate theta oscillations in a genetically engineered mouse model with light-sensitive ion channels in its GABAergic medial septum neurons. By delivering light pulses to the hippocampus, the protocol aims to synchronize neuronal activity and generate theta oscillations, which are recorded as local field potentials.

小鼠模型中 Theta 振荡的选择性作

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