这项工作是通过神经疾病研究所和中风R43 / R44 NS064661资助。

按照动物护理手术工具消毒机构的指导方针，并根据需要修改，​​以符合准则，并获得批准的机构的机构动物护理和使用委员会（IACUC）的协议。 1.手术准备<ol><li>清洁和准备发射，以确保安全和无菌手术。从防静电包装的发射器和喷雾两种或浸泡在70％乙醇。漂洗发射机用浸在无菌盐水或无菌盐水一直浸没无菌棉海绵之间的无菌盐水和地点。 </li><li>收集和消毒手术所需的工具;高压蒸汽灭菌。看表中的原料和试剂为手术工具列表。 </li></ol> 2.手术植入<ol><li>麻醉动物并根据IACUC批准的协议维持麻醉。在启动和在河畔盖瑞检查脚趾捏反射每隔15分钟。响应的缺乏表明麻醉足够的水平。 <ol><li>对于幼崽，使用麻醉异氟醚（4％）与O 2（100％）。对于成年人，用氯胺酮（100毫克/千克）与赛拉嗪（10毫克/千克）。 </li></ol></li><li>修正了立体框的位置。将耳栏提示在耳道。不要过度拧紧耳酒吧，颅骨很软的年轻的幼鼠。确保麻醉鼻锥。 <ol><li>通过将其放置在加热垫设置为37℃，保持在手术过程中的动物保暖。成年动物，适用润滑软膏动物的眼睛。 </li></ol></li><li>消毒切口部位，保持无菌手术领域。 <ol><li>拭头皮用70％乙醇和优碘的交替应用。开始于头皮的中心，并且日益广泛的同心圆。 </li><li>盖上悬垂的动物，并进行了手术过搭着动物。维持免缝勒手术野由衬外科的建立用无菌窗帘，喷射设备，用70％的乙醇。 </li><li>戴无菌手术手套和礼服（或所要求的机构）。为了帮助保持无菌区，使用手术助手。 </li></ol></li><li>使对动物的头皮稍稍落后沿中线眼睛，约2厘米的切口。插入时手术刀的头骨仍然非常柔软年轻的幼鼠谨慎使用。做一个切口使切口出血少，愈合和更快。 </li><li>暴露颅骨。准备一个干净，干燥的地方，以最大限度地提高发射机和颅骨的骨之间的纽带。用动脉瘤夹子抓头皮。 <ol><li>轻轻一拉头皮远离中线的四角。寻找解剖标志，如前囟和lambda头骨。记住头骨是不是在这个年龄段融合的动物。使用立体坐标Paxinos地图集找到正确位置钻孔。 </li></ol&gt; </li><li>使用的Dremel型工具带毛刺型钻头。创建与孔在直径上大于300微米所需的记录位置的两个颅骨钻孔。放置毛刺孔用于在颅骨的拉姆达背后小脑参考电极。 </li><li>确保发射器上的导线与毛刺的孔对齐。如果电极导线没有对准，电极的胶污染是可能的，并会导致不良的信号。要对齐电线，检查变送器的配合和轻轻弯曲电极排队超过预定的位点毛刺的孔。 </li><li>修剪电极引线。用手术剪修剪电极至所需的长度。的电极深度为所需的实验记录的类型的重要（ 即 ，放置上述硬膜的电极，用于EEG记录，或使用立体定位坐标定义的大脑结构）。 </li><li>宽松申请氰基丙烯酸酯变送器T的基础上，Ø覆盖的区域，确保避免涂层电极。氰基丙烯酸酯胶是电绝缘体，污染电极用胶水将导致无信号。 <ol><li>如果从脑深部结构的记录，安装变送器上的套管固定器，并放置在立体定位臂，用于Z轴控制。使用立体手臂合适的深度，并将氰基丙烯酸酯胶发射机围绕降低发射机。 </li></ol></li><li>放置发射器，以确保强粘合前彻底干燥的头骨。适用于发射器涂上氰基丙烯酸酯的头骨。小心对准电极与相应的颅骨钻孔。 <ol><li>尽量避免破坏性大血管结构。握住发射器的地方用一分钟轻微的压力。用轻微的压力，以形成在发射机和颅骨之间的牢固的结合。 </li></ol></li><li>施加额外氰基丙烯酸酯，足以完全密封发射器/头骨接口。为确保股份公司OOD适配性强的债券，最大限度的胶水接触头骨的表面面积。施加氰基丙烯酸酯类粘合剂在发射机一圈，确保既头骨和发射机的壁都包括在内。 </li><li>在植入发射机的基地申请通过围绕氰基丙烯酸酯类注射器化学促进剂（0.1毫升）中。使用助燃剂谨慎，小心不要申请到邻近组织。 注意:氰基丙烯酸酯固化的化学加速度确保发射机和颅骨之间的牢固的结合迅速形成。氰基丙烯酸酯促进剂是有用的粘合剂的速度固化，但不是必要的。 </li><li>通过用无菌生理盐水彻底冲洗区域除去促进剂。氰基丙烯酸酯加速器可能导致组织发炎，如果不从切口的面积洗涤。洗区域，填充1.0毫升注射器用无菌盐水，并通过注射器针头浇灌的区域。通常0.5毫升盐水足以洗出加速器。 </li><li>缝合周围的发射机的底部的皮肤，但不覆盖发射机。顶部发射机必须高于皮肤有效地传输神经信号。皮肤应该是发射器和设备周围的胶水各地合理紧。使用薇乔或丝线缝合（软线）;皮肤未成熟动物是软的，如果不使用软缝线容易损坏。对于成年动物，用任何材料缝合。 </li><li>从立体框架中取出的动物，并放置在加热毯进行恢复。 </li><li>确保动物是久违的大坝前温热（37℃）和动态（ 即完全恢复）。确保动物被捏在动物的背部皮肤（如果动物是脱水，皮肤会保持变形）水合。如果动物是脱水，管理皮下注入乳酸林格氏缓冲。不要让动物无人看管，直到它已经恢复了意识足以维持胸骨斜卧。 <ol><li>辖丁丙诺啡（0.05毫克/公斤），以动物为手术后疼痛管理和皮下注射的注射部位周围0.1毫升布比卡因。 注:从开始到结束的整个过程应在5-10分钟这个年龄（出生后6天）的动物来完成。手术时间可能需要更长的时间对老年人的动物。 </li></ol></li></ol> 3.保养和住房注意:有些大坝可能不会容忍幼仔植入设备。水坝可能需要选择谁是宽容。它是可以接受的水坝以通过拾取起来由发射机左右移动笼幼仔。 <ol><li>一旦动物断奶，单独室内它们，以避免他们的笼子队友拆除设备。 </li><li>通过致死量的戊巴比妥（25毫克/千克）或异氟烷（在钟罩）当困难的症状是本安乐死的动物。 </li><li>请注意，有些动物住房笼用铁丝插入除可FERE与植入发射机。一定要检查导线插入的高度，以确保动物不能获得发射抓住电线插入的&#39;棒&#39;之间。咨询您的兽医的帮助。 </li></ol> 4.录像脑电图<ol><li>本身或放置在一个笼子里动物共安置与同窝和大坝。然而，地方只有一个植入动物在一个单一的笼子里。不要让幼犬单独录音室中超过2小时。监测动物遇险和脱水的迹象。 </li><li>连接提供电源接收器底座和验证电源指示灯亮起。连接接收器底座使用（卡口尼尔 - Concelman）BNC电缆数据采集系统。 </li><li>将动物笼上的接收器底座（ 图2）的顶部。 "信号"灯应亮表示发射机已检测。数据现在可以被记录下来。 </li><li> ŤÓ记录数据时，接收机基连接到一个模拟-数字转换器和转换器连接到计算机（ 图1）。 </li><li>设定记录的采样率。确保数据被正确采样。记录至少选择250 Hz的采样率（推荐500赫兹）（发射机的带宽为0.1-100赫兹）。 </li><li>保存数字化数据，并使用信号处理软件如Matlab的分析。 </li></ol> 5.脑电图分析 - 通用<ol><li>执行的FFT（快速傅立叶变换）到时域脑电图数据从0-100赫兹变换到频域。 </li><li>执行从FFT使用256翰窗口段功率谱密度（PSD的）的基础上，Welch法和归由10×估计日志10（PSD）。功率谱表明主导脑电图信号在所需时间周期的特定频率。 </li><li>通过从各动物服用PSD的平均跨动物组数据随着时间的推移匹配处理。由1.96 X平均值（PSD）/平方根（n），其中n是动物数（PSD痕迹）创建的95％置信区间。绘制数据的平均值和95％置信区间，以产生脑电图跨越动物的同伙的整个频率内容的一个量化报告如比较治疗组与对照组。 </li></ol> 6.围产期缺氧缺血（HI）示范协议<ol><li>麻醉P6 -通过（从麻醉汽化器框与输入）将动物在麻醉箱用异氟烷麻醉（4％用100％O 2）的7大鼠幼鼠。在启动和手术过程中检查脚趾捏反射每隔15分钟。响应的缺乏表明麻醉足够的水平。 </li><li>将在它的后面的小狗，露出颈部和擦洗用70％乙醇和10％的优碘的交替应用。重复乙醇/优碘擦洗3次。 </li><li>制成1厘米的切口在与S颈部的皮肤cissors在颈部的中线。提起用钳子的皮肤和使用剪刀剪。小心不要让切口切开时的肌肉组织。 </li><li>使用钝性分离技术来揭露颈动脉。要执行钝性分离，使用2对钝鼻钳。插入的尖端进入组织并让外科器械的弹性作用扩散的组织。重复，直到颈动脉被暴露。一个鲜红的颜色和可视脉冲的存在确定的颈动脉。 </li><li>单独的颈动脉使用钝钳迷走神经。插入动脉和神经之间的钝尖镊子。松开钳子，并让该工具的春风行动从迷走神经分离颈动脉。 </li><li>地方动脉瘤夹4-5毫米分开放在颈动脉。小心避免快速运动不损害与夹具的动脉。 </li><li>烧灼动脉瘤夹具之间的颈动脉。烧灼动脉，触摸夹具之间的动脉热cauterizer小费。动脉被切断后，确保两端都正确烧灼，以避免出血。 </li><li>取下夹子，用3缝合关闭颈部切口。只缝合皮肤，注意不要缝合肌肉组织。 </li><li>让动物恢复1小时。监测动物的呼吸和从颈部出血。如果出血是存在，请勿使动物HI（步骤6.10）。 </li><li>放置在温度受控室中的动物，在37℃，不断地推出8％的O 2/92％N 2混合物到腔室2小时。 </li></ol>

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</ol>

博士。 Lehmkuhle和杜德克有经济利益Epitel公司，大纪元无线生物电位记录系统的设计。

它可以是非常昂贵的，使长期的脑电图记录在疾病的小动物模型。依靠简单的电路和强调低功耗，我们已经能够制造发射机系统（ 图1和2），该降低的长期监测实验的成本。 6个月的监测实验的总成本可以低至$ 470，再加上动物（〜按日1.5美元动物元，200发射器）的成本。发射器的小尺寸允许连续不间断的电摄影记录在小动物，人类疾病的临床前模型，这是非常困难的，以获得与拴系或射频的无线记录 ​​系统（ 图4）。最后，发射机的颅骨安装的性质减少了手术时间和压力上，可以以其他方式损害的实验动物。在这里，我们表现出证明型原理实验从三个差异erent实验模型发作:围产期缺氧缺血13，27，在大鼠幼鼠28（ 图4），DFP诱发癫痫持续状态（ 图5）和在遗传诱导的海绵状血管畸形模型发作（ 图6）。 可能获得无伪影，长期电图记录的最关键方面是验证不受约束电极获得的利息（ 图4-6）的皮层区。这包括公用基准/接地电极。尤其关键的是将所述发射机到所述颅骨硬膜外脑电图的应用程序。在此过程中，有可能无意中涂覆电极与腈基丙烯酸酯的前端给定电极的非常短的长度。覆盖在电极在氰基丙烯酸酯可以衰减EEG信号或完全分离它们在最坏的情况下。同样，缺乏良好的电气连接B切口白内障手术挽公共参考/地面和动物的大脑将防止差动放大器的正确操作，在发射器，从而导致在电"噪声"信号输出。通常情况下，在手术后，高质量的信号可能会受到损害，最多到48小时，由于水肿周围在头骨的毛刺的孔。随着水肿消退，信号普遍提高。这可避免通过将颅骨的表面上的电极不作毛刺的孔。这个过程的后果是增加了势涂覆电极与腈基丙烯酸酯，减小高频的活性由于颅骨的低通电特性，以及潜在的电隔离的信号中的共同参考/地呈现噪声。电极练正确放置可以用薄木板或胶合板的模仿的小鼠或大鼠颅骨的厚度来完成。在这个手稿呈现的结果说明了QUAlity可以使用无线遥测技术获得的录音。 使用本文所述的方法通过外科手术植入可以少至10分钟，这取决于手术的复杂性。用于外科进入到深部脑结构，如海马CA1区，最好是发射机连接到安装到立体定位框架上的微操作。微操作将提供外科医生用的准确性根据在小鼠29和大鼠30大脑的图谱公布立体坐标植入发射机。这可以通过简单地套结一块皮下注射针管到发射机与腈基丙烯酸酯，然后安装在显微皮下注射针来完成。的x，y和z坐标的微操纵控制将安装发射机到颅骨时缝合皮肤闭合之前，以提供额外的稳定性。周围的pe加入骨螺钉发射机rimeter可以帮助锚定发射机到头骨，虽然它们不是必要的。骨螺钉可能是有效的，然而，在发作和癫痫的某些动物模型中，如锂 - 毛果芸香碱处理的成年大鼠。这些动物往往具有自发性抽搐发作激烈的运动活动期间扣押，可能会损坏变送器。额外的复杂性可以被添加到这些实验。例如，发射机是与许多不同型号的创伤性脑损伤，如控制皮质冲击31兼容。发射机设备的耐久性通过植入动物发射机在P7和然后容纳他们在动物设施中进行测试。 12个月后，大部分的植入物保持完整的颅骨。当动物安乐死，头骨似乎是正常的和发射器被嵌入在颅骨，需要显著力来解压。请注意，当脑深部结构进行研究;作为脑的增长，并且电极保持静止，电极的最终位置将预期发生变化。对于此处所描述的技术中，电极是通常位于电极上面的硬脑膜，这使得这两个脑和头骨生长和留在原来的位置。在多久的发射机可以使用的限制因素是电池的大小（ 即 ，直到电池耗尽）。 发射器的外壳的自包含单片设计（ 即发射机被嵌入硬环氧树脂）借自己与安置与大坝及其同窝幼崽不成熟使用。通常情况下，合房植入动物有线系绳导致植入硬件或幼崽的抵消作用由坝的破坏。发射机的光滑壁形状允许植入几乎没有硬件故障或由于拆幼仔的损失。 <p claSS ="jove_content"&gt;该系统中的低功率电容耦合传输方案很适合用在许多不同的实验方案，例如放置在一个温度控制的恒温箱，甚至对于与实验装置中，如体积描记室使用。同样地，接收器天线的形状可被操纵，以适应许多不同的行为的环境，如发射机的升高的T-maze.The范围只有几英寸作为发射器必须能够"驱动器"（电容耦合到）接收机天线和依赖于动物被容纳在标准啮齿动物笼蔽或合适笼覆盖的接收机天线的表面积。发射机充当电场的一极，而该动物的身体作为另一极。该字段是使得某些取向，如90°异相的接收机天线，将无法驱动造成的接收机天线一个"脱落"。这是一个相当罕见的现象。使用这种技术今后的工作将允许来自多个动物在同一个笼子里，这将减少住宿费用，并允许社会交往记录。进一步的发展将涉及增加频道，包括温度，和肌电的数量。该设备的当前设计提供了一个用于记录经典脑电图频带，这是不适合于记录的快速波纹或高频振荡的优化的带宽。在未来，该设备可以被修改，以记录信号的高频分量，但在电池寿命的成本。不同类型的传感器将包括血压和压力容积，甚至根据期望的大脑结构的立体定向坐标构造的发射机电极布置到用户指定的尺寸。

最古老的 - 并且仍然是最广泛使用的 - 技术在大脑中记录生物电势是脑电图（EEG）。它是临床上用于神经系统异常，包括扣押检测1，癫痫病灶2的定位，和3,4震荡诊断。这种技术也被广泛地用于提供关于睡眠的机制的基本信息和诊断睡眠障碍5,6。 如在癫痫的临床诊断，脑电图已成为不可缺少的转化研究在遗传和获得性癫痫的动物模型。在目前的研究应用，"有线"或"拴"录音是标准，而在成年老鼠在一个时间7例行的几个星期。然而，电噪声，运动伪影，并拴动物会伤害自己通过拉电缆的风险早已COMPRomised这些实验。因此，为了改善实验条件和成功率，我们需要开发新的技术，将允许用于消除动物和仪器之间的有线接口。发展的最明显的区是设计和实现遥测系统，其允许高品质的录音，同时保持了长的使用寿命和减少不适用于动物受试者的。减少这些设备的物理尺寸将使转化研究神经障碍的新生儿和少年啮齿动物模型。 低通道数的脑电图记录大鼠被广泛用来开发新的疗法来抑制癫痫发作能够翻译给人类。从一个或多个站点进行长时间录像打开许多可能性，使用癫痫的啮齿动物模型中的转化研究。大部分的当代研究在这一领域的目标是阻止慢性seiz的发生数目字或癫痫（ 即癫痫）的发展，这样的研究工作需要大量的，如果不连续脑电图监测，以测定所提出的治疗8的有效性;一个小的，简单的，遥测系统，一个，两个或四个通道每通道0.1-100赫兹之间运行将有力地推动这种类型的转化研究。电图发作常常发生以最小的行为（当然无抽搐），这限制了基于行为发作测定的有用性。结合脑电图记录和同步视频监控的策略，允许一个捕捉每一个癫痫发作的可能性;进而，这些分析方法可能会允许发生的"发作"（或扣押）事件9间脑癫痫发作的尖峰的定量评估。此外，为了获得连续的高品质低工件脑电图记录的能力，为此，无线技术是通常优越，将允许使用的基于计算机的算法开发用于研究特定的脑电图波形（ 例如 ，θ-，γ），以及自动检测癫痫发作，显著减少实验者的工作量。 初级临床前模型为脑损伤后研究慢性癫痫是成年大鼠或小鼠，无论是通过一个化学惊厥（ 即 ，红藻氨酸或毛果芸香碱）或电诱导癫痫持续状态（SE），其后是慢性癫痫。在这些条件下，与SE或在癫痫动物随后发作相关的严重惊厥可导致损伤从动物撕裂或拉系绳和松开该保持headcap的连接的螺钉。最终，正是这种问题，通常终止这些实验，但需要获得长期高分辨率脑电图记录实验旨在开发用于治疗慢性新疗法癫痫是最重要的。此外，住房，监控和分析从长期植入动物的数据是在两个直接成本和研究者时间一个相当大的投资;因此，提前终止实验可能导致显著成本研究人员。由于癫痫病的进展，这些车型中，通常发作更为频繁和更严重的10-12，增加了动物受伤的可能性，正如其效用开发新的治疗变得最大。这些动物通常可以开发数十种每天抽搐发作的，在集群经常13发生。 大概在生物医学科学的最重要的发展之一是利用基因在小鼠模型定位。这种方法已允许的，并且将继续允许，遗传性癫痫的动物模型的发展即重现实际人类综合征14-16。遗传操作，可以作为进行证明性的原则疗法来抑制癫痫发作，甚至阻止癫痫的脑损伤后17-20发展。这种类型的研究将从执行脑电图高通量的连续记录能力显着受益。目前，有可能从小鼠或者拴系或遥测系统来记录;然而，获得高品质的挑战，无瑕疵的录音基本上比老鼠更困难，而且往往这需要各种形式的背包，老鼠一直试图消除。应力可能会增加癫痫发作严重性，频率和/或持续时间，从而最终将修改的实验动物的癫痫，从而混杂的研究。小，重量轻，低轮廓微型遥测系统将有利于长期脑电图从人类疾病的遗传小鼠模型的记录。 除了上述问题，记录脑电图未成熟啮齿动物模型疾病S有它自己的一套独特的挑战。未成熟动物重达一点在6克（P8鼠标）17克（P6大鼠）。这几乎是不可能使从系绳和无力串行多天拴脑电图记录由于增加的应力，以允许小狗由坝的自然饲养。直到动物断奶，他们必须留在大坝的照顾。大坝是容易摧毁任何外在连接器组件上的小狗，终止小狗，在某些情况下终止整个垃圾。此外，未成熟的啮齿动物颅骨使得难以安装任何电极座与机械完整性的​​头骨。这些挑战，唯一的啮齿动物不成熟，需要制作稳健，长期的脑电图记录了一种新的解决方案。在这里，我们专注于使用一种新型的微型无线发射器和证明性的原则，目前三个实验作为例子使用的微型无线遥测系统展示脑电图的植入和记录:1）的IM缺氧缺血大鼠成熟模型小狗，2）与DFP成年小鼠治疗诱发癫痫持续状态和随后的自发性发作，以及各种可导致癫痫发作和死亡的成年小鼠血管海绵状血管瘤3）遗传模型。 微型无线遥测系统被设计为满足四个主要要求:（1）微创手术植入; （2）对啮齿动物幼仔的大坝和同窝住房的兼容性;的单元（3）低的功耗，因此允许连续监测无手术重新植入月; （四）能够录制高品质的脑电波形​​，最小的运动伪影。无线发射机重&lt;0.6，2.3，和4克，并且&lt;0.3，0.8和1.4 cm 3的根据电池用足迹的5×7,7×9，或7×12毫米，而且容易安装到颅骨用氰基丙烯酸酯凝胶的动物。不需要骨钉锚设备安全地固定到颅骨，减少需要被钻在颅骨和手术时间孔的数量。该装置能够放大两个通道脑电图或局部场电位的从脑深部结构，如海马，超过2周，2个月，或6个月在此配置的。无线发射器的小尺寸减少感染的风险，提高了动物的流动性，并最终降低了发病率和死亡率，否则会增加的时间，金钱和数目所需要的实验动物。所有的电子设备和电池均封装在医疗级环氧树脂，使装置防水，坚韧，防止堤坝咀嚼，否则会使得设备无法使用的发射器。不像射频发射机，所述遥测系统采用的发射机和接收机天线，它位于所述动物笼子下面，允许用户保持动物在标准啮齿动物壳体之间的电容耦合。 recordi的多个通道纳克允许多模态生物电势，如心电图和脑电图记录。共患疾病的动物模型将行为21-23时记录生物电势的能力中受益。用脑电图监测相结合的行为会为研究人员提供研究和临床前研究一个更好的工具。

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long-term continuous eeg monitoring in small rodent models of human disease using the epoch wireless transmitter system

许多进步神经系统疾病在人类中，如癫痫，需要临床前的动物模型是缓慢发展的疾病，以测试干预在疾病过程的各个阶段。这些动物模型尤其难以实现不成熟的啮齿动物，一个经典的模式生物对这些疾病的实验室研究。连续记录脑电图癫痫发作等神经系统疾病的年轻动物模型提出了一个技术挑战，由于年轻老鼠的小的物理尺寸和其前断奶的水坝的依赖。因此，不仅是显然需要改善的临床前研究，这将更好地识别适于翻译到诊所，但还需要一种能够在未成熟啮齿类记录连续脑电图的新设备的那些疗法。在这里，我们描述了技术落后和演示使用了一种新的微型遥测系统，专门设计用于在幼鼠o使用ř小鼠，这也是有效的成年动物的使用。

我们制定并实施从单一的成人啮齿动物，系统化如图1脑电图记录的理念，为IACUC审批过程中，设计必须很好地集成到现有的体制动物设施。因此，该系统的设计可以很容易地在不使用的额外的空间安装在标准的动物设施:动物被容纳在一个普通的"动物设施-问题"住房笼被放置在一个接收器的具有集成的法拉第笼，以减少电噪声。来自每个接收器的碱信号由导线连接到一个计算机（ 图1）一个数字化仪进行。一台计算机是需要从同时录制，这取决于用户的数据采集系统的能力达32动物收集数据。这种类型的设置功耗很低，并产生很少的热量，气候控制动物设施兼容的功能。数据可以是外部硬盘驱动器显示在监视器上进行实时，允许试验监测，并存储长期（10 TB的存储单元）。 为了减少损失由垃圾的队友和小狗拆的坝，我们测试了各种发射器的外形。最终的设计是一个半球形的缸;形状难以鼠咬和损坏。单个发射机上的成年大鼠的颅骨示于图2A和高密度（32动物）接收机的碱，并且其中标准啮齿动物壳体放置记录钻机的早期版本中示出在图2B中。电源效率是一个非常重要的考虑因素;我们选择了电容耦合作​​为数据传输协议。下面的设计允许连续记录脑电图6个月以上视电池容量（ 图2A）。小鼠年仅12日龄（P12， 图3A）和大鼠年仅P6（FIGURË3B）容忍发射机相当不错。附着发射机与氰基丙烯酸酯颅骨使动物与发射机成长到成年（ 图3C），同时保持连续采集的脑电图数据。 发射器和无线接口的微型独特外形借给本身与新 - 和围产期疾病动物模型的工作。在图4中的数据示出了两个信道的亚急性发作活动的脑电图记录遵循缺氧缺血（HI）梗塞（颈动脉结扎，接着通过2小时缺氧用8％O 2的混合物）在一个P7的Sprague-Dawley的大鼠小狗13。该HI处理引起的大病灶半球同侧颈动脉结扎。在这里，录音显示了两种广义发作过受伤的大脑两个半球集群。黑色的曲线描绘在对侧半球脑电活动的病变部位，蓝色迹线示出了在同侧半球（ 即在病灶的面积）脑电图。而癫痫发作是存在于大脑的两个半球，该半球ispilateral脑电图显示背景抑制，这表明正在进行的脑损伤21。 癫痫持续状态可通过注入动物与有机磷酸酯，DFP 22,23被诱导成年大鼠。在图5中示出的重复EEG放电的数据，这是指示癫痫持续状态（见颞扩展图5A，B）。下面的示例痕迹，癫痫持续状态的时间过程超过12小时已经分析了非线性混合效应模型，量化癫痫发作的强度随着时间的推移。癫痫持续状态的严重程度是通过脑电图功率在伽玛频带（20-60赫兹）中所定义。在这里，上述功率平均在12个动物，并绘制了12小时与95％置信区间。钍<html> Ë数据显示DFP处理，这仍然超过12小时期间该动物连续监测的第一个小时内以伽玛功率显着增加。分析的下列方法可以用于治疗急性癫痫持续状态的严重程度的定量测量，这种现象以前主要使用行为的措施进行分析。我们包括此分析技术，例如，因为它利用了经典的EEG频带功率计算，并已广泛用于临床前研究为测试抗惊厥药物的功效在我们的实验室24-26。可能使连续的，不间断的无线录音用无线遥测的最有价值的方面是记录的低发病率发生异常自发事件的能力。这些类型的数据表明无线发射机系统的广泛的实用性。 554fig1.jpg"/&gt; 图1:历元记录系统的原理图的无线记录 ​​系统由两部分组成:1）的无线颅骨安装的发射器，其放大该生物信号，以及2）一个接收器板置于下面标准啮齿动物壳体。接收机碱的输出是由解调的生物信号放大到最多4 V峰 - 峰模拟信号。这个信号然后可以送入用于记录一个数据采集系统。 <img alt="图2" src="/files/ftp_upload/52554/52554fig2.jpg" /> 图2:发射器和接收这种特殊的无线发射器（A）的重4 g且位移&lt;1.4厘米3体积和很容易安装到大鼠和小鼠的颅骨的具有7×12毫米的足迹。发射机可以放大2通道生物电势的时间长达6个月后，其中电池是漏ED。较大的电池可用于较长的记录时间。动物被放置在标准啮齿动物笼蔽上的时代接收机（B）的顶部。右图所示是两个单独的记录钻机各自能够从16只记录同时表明每个记录钻机的相对小的足迹（2&#39;×4&#39;，约60厘米×120厘米）的一个早期例子。 <img alt="图3" src="/files/ftp_upload/52554/52554fig3.jpg" /> 图3:植入无线发射器在大鼠和小鼠的发射器可以连续脑电图记录长达6个月的小鼠年仅产后12天（P12，顶部）。中间的照片是一个P7大鼠小狗植入微型发射器。该发射器固定牢固地附着在头骨作为动物的成熟。在底部的动物是P280，被植入在年龄P7是假的发射器。该系统使simulta年龄从多个动物neous和连续脑电图记录P7通过断奶，减少所需的临床前，长期，脑电图监测研究凋落物的数量。 <img alt="图4" src="/files/ftp_upload/52554/52554fig4.jpg" /> 图4:双声道录音缺血缺氧引起的癫痫发作与遥测系统脑电图异常的双声道录音过程中有8％的O 2处理（缺氧）的颈动脉结扎（缺血）后P7大鼠小狗的无线遥测。 （A）和（B），扩大了意见波形。癫痫发作是存在于两个半球（黑，蓝）有大量的脑电图抑制目前在缺血性梗死（蓝色）半球。 <img alt="图5" src="/files/ftp_upload/52554/52554fig5.jpg" /> 图5:录制癫痫状态CUS在成年鼠。表面脑电图记录（ 即硬脑膜）与微型无线遥测系统响应二异丙基在成年大鼠（DFP）的治疗。在顶部曲线（A和B）的阴影周期将在下面的痕迹扩大享有波形。记录有无线发射机数据然后可以在频域，允许在动物的队列统计比较进行分析。 （C）的数据是伽玛带电源（20-60赫兹）以下DFP诱发癫痫持续状态（N = 12）在DFP给药后12小时平均值和95％置信区间。 <img alt="图6" src="/files/ftp_upload/52554/52554fig6.jpg" /> 图6:癫痫发作和变化脑电图血管海绵状血管瘤的转基因小鼠模型的记录在这里，我们从转基因小鼠记录（A） 的第在正在经历癫痫发作。首先，正常脑电图图形存在（1）;之前立即发作有一段预发作抑郁（2），之后是由五个发作（3）集群。以下的癫痫，异常发作放电存在于信号（4）。控制动物无癫痫发作和无异常脑电图特点（B）。

Watch this Scientific Journal Video about Long-term Continuous EEG Monitoring in Small Rodent Models of Human Disease Using the Epoch Wireless Transmitter System at JoVE.com

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

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 ...

long-term-continuous-eeg-monitoring-small-rodent-models-human-disease

Research

JoVE Journal

Neuroscience

25.5K 次观看.  Yale University School of Medicine. 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.

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

Simultaneous Video-EEG-ECG Monitoring to Identify Neurocardiac Dysfunction in Mouse Models of Epilepsy

In epilepsy, seizures can evoke cardiac rhythm disturbances such as heart rate changes, conduction blocks, asystoles, and arrhythmias, which can potentially increase risk of sudden unexpected death in epilepsy (SUDEP). Electroencephalography (EEG) and electrocardiography (ECG) are widely used clinical diagnostic tools to monitor for abnormal brain and cardiac rhythms in patients. Here, a technique to simultaneously record video, EEG, and ECG in mice to measure behavior, brain, and cardiac activities, respectively, is described. The technique described herein utilizes a tethered (i.e., wired) recording configuration in which the implanted electrode on the head of the mouse is hard-wired to the recording equipment. Compared to wireless telemetry recording systems, the tethered arrangement possesses several technical advantages such as a greater possible number of channels for recording EEG or other biopotentials; lower electrode costs; and greater frequency bandwidth (i.e., sampling rate) of recordings. The basics of this technique can also be easily modified to accommodate recording other biosignals, such as electromyography (EMG) or plethysmography for assessment of muscle and respiratory activity, respectively. In addition to describing how to perform the EEG-ECG recordings, we also detail methods to quantify the resulting data for seizures, EEG spectral power, cardiac function, and heart rate variability, which we demonstrate in an example experiment using a mouse with epilepsy due to Kcna1 gene deletion. Video-EEG-ECG monitoring in mouse models of epilepsy or other neurological disease provides a powerful tool to identify dysfunction at the level of the brain, heart, or brain-heart interactions.

Here, we present a protocol to record brain and heart bio signals in mice using simultaneous video, electroencephalography (EEG), and electrocardiography (ECG). We also describe methods to analyze the resulting EEG-ECG recordings for seizures, EEG spectral power, cardiac function, and heart rate variability.

同步视频脑电图-心电图监测对小鼠癫痫模型 Neurocardiac 功能障碍的鉴别

In epilepsy, seizures can evoke cardiac rhythm disturbances such as heart rate changes, conduction blocks, asystoles, and arrhythmias, which can ...

科研

遗传学

Noninvasive, High-throughput Determination of Sleep Duration in Rodents

Traditionally, sleep is monitored by an electroencephalogram (EEG). EEG studies in rodents require surgical implantation of the electrodes followed by a long recovery period. To perform an EEG recording, the animal is connected to a receiver, creating an unnatural tether to the head-mount. EEG monitoring is time consuming, carries risk to the animal, and is not a completely natural setting for the measurement of sleep. Alternative methods to detect sleep, particularly in a high-throughput fashion, would greatly advance the field of sleep research. Here, we describe a validated method for detecting sleep via activity-based home-cage monitoring. Previous studies have shown that sleep assessed via this method has a high degree of agreement with sleep defined by traditional EEG-based measures. Whereas this method is validated for total sleep time, it is important to note that sleep bout duration should be assessed by an EEG which has better temporal resolution. The EEG can also differentiate rapid eye movement (REM) and non-REM sleep, giving more detail about the exact nature of sleep. Nevertheless, activity-based sleep determination can be used to analyze multiple days of undisturbed sleep and to assess sleep as a response to an acute event (like stress). Here, we show the power of this system to detect the response of mice to daily intraperitoneal injections.

We describe a high-throughput method of measuring sleep by means of activity-based home-cage monitoring. This method offers advantages over traditional EEG-based methods. It is well validated for the determination of total sleep duration and can be a powerful tool to monitor sleep in rodent models of human disease.

无创高通量测定啮齿类动物睡眠时间

Traditionally, sleep is monitored by an electroencephalogram (EEG). EEG studies in rodents require surgical implantation of the electrodes followed by a ...

行为学

Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenetic Manipulation of Neuronal Activity in the Rodent Cerebral Cortex

Although the brain represents less than 5% of the body by mass, it utilizes approximately one quarter of the glucose used by the body at rest1. The function of non rapid eye movement sleep (NREMS), the largest portion of sleep by time, is uncertain. However, one salient feature of NREMS is a significant reduction in the rate of cerebral glucose utilization relative to wakefulness2-4. This and other findings have led to the widely held belief that sleep serves a function related to cerebral metabolism. Yet, the mechanisms underlying the reduction in cerebral glucose metabolism during NREMS remain to be elucidated.
	One phenomenon associated with NREMS that might impact cerebral metabolic rate is the occurrence of slow waves, oscillations at frequencies less than 4 Hz, in the electroencephalogram5,6. These slow waves detected at the level of the skull or cerebral cortical surface reflect the oscillations of underlying neurons between a depolarized/up state and a hyperpolarized/down state7. During the down state, cells do not undergo action potentials for intervals of up to several hundred milliseconds. Restoration of ionic concentration gradients subsequent to action potentials represents a significant metabolic load on the cell8; absence of action potentials during down states associated with NREMS may contribute to reduced metabolism relative to wake.
	Two technical challenges had to be addressed in order for this hypothetical relationship to be tested. First, it was necessary to measure cerebral glycolytic metabolism with a temporal resolution reflective of the dynamics of the cerebral EEG (that is, over seconds rather than minutes). To do so, we measured the concentration of lactate, the product of aerobic glycolysis, and therefore a readout of the rate of glucose metabolism in the brains of mice. Lactate was measured using a lactate oxidase based real time sensor embedded in the frontal cortex. The sensing mechanism consists of a platinum-iridium electrode surrounded by a layer of lactate oxidase molecules. Metabolism of lactate by lactate oxidase produces hydrogen peroxide, which produces a current in the platinum-iridium electrode. So a ramping up of cerebral glycolysis provides an increase in the concentration of substrate for lactate oxidase, which then is reflected in increased current at the sensing electrode. It was additionally necessary to measure these variables while manipulating the excitability of the cerebral cortex, in order to isolate this variable from other facets of NREMS.
	We devised an experimental system for simultaneous measurement of neuronal activity via the elecetroencephalogram, measurement of glycolytic flux via a lactate biosensor, and manipulation of cerebral cortical neuronal activity via optogenetic activation of pyramidal neurons. We have utilized this system to document the relationship between sleep-related electroencephalographic waveforms and the moment-to-moment dynamics of lactate concentration in the cerebral cortex. The protocol may be useful for any individual interested in studying, in freely behaving rodents, the relationship between neuronal activity measured at the electroencephalographic level and cellular energetics within the brain.

A procedure is described for manipulating the activity of cerebral cortical pyramidal neurons optogenetically while the electroencephalogram, electromyogram, and cerebral lactate concentration are monitored. Experimental recordings are performed on cable-tethered mice while they undergo spontaneous sleep/wake cycles. Optogenetic equipment is assembled in our laboratory; recording equipment is commercially available.

同时脑电图，实时测量的啮齿动物的大脑皮质中的神经元活动的乳酸浓度与Optogenetic的操作

Although the brain represents less than 5% of the body  by mass, it utilizes approximately one quarter of the glucose used by the body  at rest1. The ...

神经科学

Use of a Wireless Video-EEG System to Monitor Epileptiform Discharges Following Lateral Fluid-Percussion Induced Traumatic Brain Injury

The lateral fluid percussion injury (FPI) model is well established and has been used to study TBI and post-traumatic epilepsy (PTE). However, considerable variability has been reported for the specific parameters used in different studies that have employed this model, making it difficult to harmonize and interpret the results between laboratories. For example, variability has been reported regarding the size and location of the craniectomy, how the Luer lock hub is placed relative to the craniectomy, the atmospheric pressure applied to the dura and the duration of the pressure pulse. Each of these parameters can impact injury severity, which directly correlates with the incidence of PTE. This has been manifested as a wide range of mortality rates, righting reflex times and incidence of convulsive seizures reported. Here we provide a detailed protocol for the method we have used to help facilitate harmonization between studies. We used FPI in combination with a wireless EEG telemetry system to continuously monitor for electrographic changes and detect seizure activity.&#160; FPI is induced by creating a 5 mm craniectomy over the left hemisphere, between the Bregma and Lambda and adjacent to the lateral ridge. A Luer lock hub is secured onto the skull over the craniectomy. This hub is connected to the FPI device, and a 20-millisecond pressure pulse is delivered directly to the intact dura through pressure tubing connected to the hub via a twist lock connector. Following recovery, rats are re-anesthetized to remove the hub. Five 0.5 mm, stainless steel EEG electrode screws are placed in contact with the dura through the skull and serve as four recording electrodes and one reference electrode. The electrode wires are collected into a pedestal connector which is secured into place with bone cement. Continuous video/EEG recordings are collected for up to 4 weeks post TBI.

Here we present a protocol to induce severe TBI with the lateral fluid percussion injury (FPI) model in adult, male Wistar rats. We also demonstrate the use of a wireless telemetry system to collect continuous video-EEG recordings and monitor for epileptiform discharges consistent with post-traumatic epileptogenesis.&#160;

使用无线视频-EEG系统监测侧流冲击后癫痫的放电诱发创伤性脑损伤

The lateral fluid percussion injury (FPI) model is well established and has been used to study TBI and post-traumatic epilepsy (PTE). However, ...

Simultaneous Monitoring of Wireless Electrophysiology and Memory Behavioral Test as a Tool to Study Hippocampal Neurogenesis

Brainwaves amplitude obtained from electroencephalography (EEG) has been well-recognized as a basis for cognitive capacity, memory, and learning on animals and humans. Adult neurogenesis mechanism is also linked to memory and learning improvement. Traditionally, researchers used to assess learning and memory parameters in rodent models by behavioral tasks. Therefore, the simultaneous monitoring of behavioral changes and EEG is particularly interesting in correlating data between brain activity and task-related behaviors. However, most of the equipment required to perform both studies are either complex, expensive, or uses a wired setup network that hinders the natural animals&#8217; movement. In this study, EEG was recorded with a wireless electrophysiology device along with the execution of a novel object recognition task (NORT). The animal&#8217;s behavior was monitored simultaneously by a video tracking system. Both recordings were analyzed offline by their timestamps which were synched to link EEG signals with the animal&#8217;s actions. Subjects consist of adult Wistar rats after medium-term environmental enrichment treatment. Six skull screw electrodes were fixed in pairs on both hemispheres over frontal, central, and parietal regions and were referenced to an electrode located posterior of the nasal bone. NORT protocol consists of exposing the animal to two identical objects for 10 min. After 2 h and 24 h, one of the objects was replaced with a novel one. Exploration time for each object was monitored by a behavioral tracking software (BTS) and EEG data recording. The analysis of the EEG synced with behavioral data consists of estimations of alpha and beta relative band power and comparisons between novel object recognition versus familiar object exploration, between three experimental stages. In this manuscript, we have discussed electrodes manufacturing process, epidural electrodes implantation surgery, environmental enrichment protocol, NORT protocol, BTS setup, EEG &#8211; BTS coupling for simultaneous monitoring in real-time, and EEG data analysis based on automatic events detection.

The protocol presented here provides information on the simultaneous electroencephalography (EEG) and behavioral assessment in real-time. We have discussed all steps involved in this protocol as an attractive solution for researchers in many fields of neuroscience, particularly in learning and memory areas.

同时监测无线电生理学和记忆行为测试作为研究海马神经发生的工具

Brainwaves amplitude obtained from electroencephalography (EEG) has been well-recognized as a basis for cognitive capacity, memory, and learning on ...

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 ...

High-density Electroencephalographic Acquisition in a Rodent Model Using Low-cost and Open-source Resources

Advanced electroencephalographic analysis techniques requiring high spatial resolution, including electrical source imaging and measures of network connectivity, are applicable to an expanding variety of questions in neuroscience. Performing these kinds of analyses in a rodent model requires higher electrode density than traditional screw electrodes can accomplish. While higher-density electroencephalographic montages for rodents exist, they are of limited availability to most researchers, are not robust enough for repeated experiments over an extended period of time, or are limited to use in anesthetized rodents.1-3 A proposed low-cost method for constructing a durable, high-count, transcranial electrode array, consisting of bilaterally implantable headpieces is investigated as a means to perform advanced electroencephalogram analyses in mice or rats.

Procedures for headpiece fabrication and surgical implantation necessary to produce high signal to noise, low-impedance electroencephalographic and electromyographic signals are presented. While the methodology is useful in both rats and mice, this manuscript focuses on the more challenging implementation for the smaller mouse skull. Freely moving mice are only tethered to cables via a common adapter during recording. One version of this electrode system that includes 26 electroencephalographic channels and 4 electromyographic channels is described below.

Instructions for the low-cost construction and surgical implantation of a chronic transcranial high-density electroencephalographic montage into mice are provided. Signal recording, extraction, and processing techniques are also described.

高密度的脑电图采集在啮齿动物模型使用低成本的开源资源

Advanced electroencephalographic analysis techniques requiring high spatial resolution, including electrical source imaging and measures of network ...

The Pilocarpine Model of Temporal Lobe Epilepsy and EEG Monitoring Using Radiotelemetry System in Mice

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status epilepticus (SE) is frequently selected to recapitulate spontaneous recurrent seizures (SRS). Here we present a protocol of SE induction by intraperitoneal (i.p.) injection of pilocarpine and monitoring of chronic recurring seizures in live animals using a wireless telemetry video and electroencephalogram (EEG) system. We demonstrated notable behavioral changes that need attention after pilocarpine injection and their correlation with hippocampal neuronal loss at 7 days and 6 weeks post-pilocarpine. We also describe the experimental procedures of electrode implantation for video and EEG recording, and analysis of the frequency and duration of chronic recurrent seizures. Finally, we discuss the possible reasons why the expected results are not achieved in each case. This provides a basic overview of modeling chronic epilepsy in mice and guidelines for troubleshooting. We believe this protocol can serve as a baseline for suitable models of chronic epilepsy and epileptogenesis.

This manuscript describes a method of inducing status epilepticus by systemic pilocarpine injection and of monitoring spontaneous recurrent seizures in live animals using a wireless telemetry video and electroencephalogram system. This protocol can be utilized for studying the pathophysiologic mechanisms of chronic epilepsy, epileptogenesis, and acute seizures.

小鼠颞叶癫痫的 Radiotelemetry 模型及脑电图监测

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status ...

医学

<meta charset="utf8"></head><body>使用皮下电极放置的兔子连续 EEG/ECG 记录

Surgical Implant Procedure and Wiring Configuration for Continuous Long-Term EEG/ECG Monitoring in Rabbits

A multi-system approach that includes the continuous collection of electroencephalogram (EEG) and electrocardiogram (ECG) recordings facilitates a comprehensive assessment of electrical activity in the brain and heart, particularly leading up to and surrounding episodic events. Building on our previous protocol that facilitates the collection of intermittent EEG/ECG recordings from conscious restrained rabbits, we developed a protocol for the collection of continuous EEG/ECG recordings from unrestrained rabbits. This new method enables high-quality 24/7 recordings in the housing cage, which captures the normal range of physiological states and enables the rabbits to move, eat, drink, and sleep freely. It enables comprehensive assessments of the prevalence, incidence, and susceptibility to EEG/ECG abnormalities, seizures, arrhythmias, and sudden death. This procedure involves the surgical placement of subdermal electrodes and the design and manufacturing of a robust wiring system that is impervious to damage by the rabbits. The surgical procedure includes two incisions and the sub-dermal tunneling of nine electrodes and wires that exit through a custom-made port. The external connector attaches to a wire harness that includes an electrical swivel, which allows for&#160;free range of rabbit motion throughout its cage. Additionally, the wires are enclosed in a metal sheath suspended by a retractor cable from the ceiling of the rabbit&#39;s cage. The wires are then connected to an amplifier/digitizer for the acquisition of high-quality continuous EEG/ECG recordings.

<meta charset="utf8"></head><body>用于兔连续长期 EEG/ECG 监测的外科植入物手术和布线配置

The study outlines a protocol for the design, manufacturing, and placement of a novel continuous EEG/ECG recording system in rabbits. This hard-wired system allows for continuous EEG/ECG recordings from nine distinct electrodes on the head and chest, which facilitates comprehensive assessments of electrical activity in the brain and heart.