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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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。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Sterile Surgical Gloves</td>
			<td>Protective Industrial Products</td>
			<td>100-3201 PF</td>
			<td>Power Free Sterile Latex Surgical Glove</td>
		</tr>
		<tr>
			<td>Scalpel Handle</td>
			<td>FST</td>
			<td>10003-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel Blade #15</td>
			<td>FST</td>
			<td>10015-00</td>
			<td></td>
		</tr>
		<tr>
			<td>Fine Scissors</td>
			<td>FST</td>
			<td>14090-09</td>
			<td></td>
		</tr>
		<tr>
			<td>Burr tool</td>
			<td>Ram Products, Inc.</td>
			<td>Microtorque II</td>
			<td></td>
		</tr>
		<tr>
			<td>Fine burr</td>
			<td>FST</td>
			<td>19007-07</td>
			<td></td>
		</tr>
		<tr>
			<td>Aneurism clip</td>
			<td>ROBOZ</td>
			<td>RS-5422</td>
			<td></td>
		</tr>
		<tr>
			<td>Toothed Forceps</td>
			<td>FST</td>
			<td>11022-14</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton-Tipped applicators</td>
			<td>McKesson</td>
			<td>24-103</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle Driver</td>
			<td>WPI</td>
			<td align="right">521725</td>
			<td>Olsen-Hegar Needle Holder</td>
		</tr>
		<tr>
			<td>Cyanoacrylate gel</td>
			<td>Henkel</td>
			<td>Loctite 4541</td>
			<td></td>
		</tr>
		<tr>
			<td>Cyanoacrylate accelerant</td>
			<td>Henkel</td>
			<td>Loctite 7452</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture</td>
			<td>Ethicon</td>
			<td colspan="2">Vicryl RB-1 J304</td>
		</tr>
		<tr>
			<td>Elecrocautery disposable</td>
			<td>Bovie</td>
			<td>AA01</td>
			<td>Fine Tip</td>
		</tr>
		<tr>
			<td>Surgical Tray</td>
			<td>FST</td>
			<td>20311-21</td>
			<td></td>
		</tr>
		<tr>
			<td>Epitel Receiver Base</td>
			<td>Epitel Inc</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Epitel wireless transmitter</td>
			<td>Epitel Inc</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Biopac digitizer</td>
			<td>Biopac</td>
			<td>MP-150</td>
			<td></td>
		</tr>
		<tr>
			<td>PC-compatible computer</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

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

25.4K 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

Systemic and Local Drug Delivery for Treating Diseases of the Central Nervous System in Rodent Models

Thorough preclinical testing of central nervous system (CNS) therapeutics includes a consideration of routes of administration and agent biodistribution in assessing therapeutic efficacy. Between the two major classifications of administration, local vs. systemic, systemic delivery approaches are often preferred due to ease of administration. However, systemic delivery may result in suboptimal drug concentration being achieved in the CNS, and lead to erroneous conclusions regarding agent efficacy. Local drug delivery methods are more invasive, but may be necessary to achieve therapeutic CNS drug levels. Here, we demonstrate proper technique for three routes of systemic drug delivery: intravenous injection, intraperitoneal injection, and oral gavage. In addition, we show a method for local delivery to the brain: convection-enhanced delivery (CED). The use of fluorescently-labeled compounds is included for in vivo imaging and verification of proper drug administration. The methods are presented using murine models, but can easily be adapted for use in rats.

Thorough preclinical testing of drugs that act in the central nervous system often involves assessing and comparing drug biodistribution in association with specific routes of administration. Here, three commonly used methods of systemic delivery (intravenous, intraperitoneal, and oral) as well as a method for local delivery (convection-enhanced delivery) are demonstrated in mice.

全身和局部给药治疗的中枢神经系统疾病的啮齿动物模型

Thorough preclinical testing of central nervous system (CNS) therapeutics includes a consideration of routes of administration and agent biodistribution ...

科研

神经科学

Efficient Derivation of Human Neuronal Progenitors and Neurons from Pluripotent Human Embryonic Stem Cells with Small Molecule Induction

There is a large unfulfilled need for a clinically-suitable human neuronal cell source for repair or regeneration of the damaged central nervous system (CNS) structure and circuitry in today's healthcare industry. Cell-based therapies hold great promise to restore the lost nerve tissue and function for CNS disorders. However, cell therapies based on CNS-derived neural stem cells have encountered supply restriction and difficulty to use in the clinical setting due to their limited expansion ability in culture and failing plasticity after extensive passaging1-3. Despite some beneficial outcomes, the CNS-derived human neural stem cells (hNSCs) appear to exert their therapeutic effects primarily by their non-neuronal progenies through producing trophic and neuroprotective molecules to rescue the endogenous cells1-3. Alternatively, pluripotent human embryonic stem cells (hESCs) proffer cures for a wide range of neurological disorders by supplying the diversity of human neuronal cell types in the developing CNS for regeneration1,4-7. However, how to channel the wide differentiation potential of pluripotent hESCs efficiently and predictably to a desired phenotype has been a major challenge for both developmental study and clinical translation. Conventional approaches rely on multi-lineage inclination of pluripotent cells through spontaneous germ layer differentiation, resulting in inefficient and uncontrollable lineage-commitment that is often followed by phenotypic heterogeneity and instability, hence, a high risk of tumorigenicity7-10. In addition, undefined foreign/animal biological supplements and/or feeders that have typically been used for the isolation, expansion, and differentiation of hESCs may make direct use of such cell-specialized grafts in patients problematic11-13. To overcome these obstacles, we have resolved the elements of a defined culture system necessary and sufficient for sustaining the epiblast pluripotence of hESCs, serving as a platform for de novo derivation of clinically-suitable hESCs and effectively directing such hESCs uniformly towards clinically-relevant lineages by small molecules14 (please see a schematic in Fig. 1). Retinoic acid (RA) does not induce neuronal differentiation of undifferentiated hESCs maintained on feeders1, 14. And unlike mouse ESCs, treating hESC-differentiated embryoid bodies (EBs) only slightly increases the low yield of neurons1, 14, 15. However, after screening a variety of small molecules and growth factors, we found that such defined conditions rendered retinoic acid (RA) sufficient to induce the specification of neuroectoderm direct from pluripotent hESCs that further progressed to neuroblasts that generated human neuronal progenitors and neurons in the developing CNS with high efficiency (Fig. 2). We defined conditions for induction of neuroblasts direct from pluripotent hESCs without an intervening multi-lineage embryoid body stage, enabling well-controlled efficient derivation of a large supply of human neuronal cells across the spectrum of developmental stages for cell-based therapeutics.

We have established a protocol for induction of neuroblasts direct from pluripotent human embryonic stem cells maintained under defined conditions with small molecules, which enables derivation of a large supply of human neuronal progenitors and neuronal cell types in the developing CNS for neural repair.

人类神经祖细胞和神经元小分子诱导人类胚胎干细胞的多能干高效的推导

There is a large unfulfilled need for a clinically-suitable human neuronal cell source for repair or regeneration of the damaged central nervous system ...

A Molecular Readout of Long-term Olfactory Adaptation in C. elegans

During sustained stimulation most sensory neurons will adapt their response by decreasing their sensitivity to the signal. The adaptation response helps shape attention and also protects cells from over-stimulation. Adaptation within the olfactory circuit of C. elegans was first described by Colbert and Bargmann1,2. Here, the authors defined parameters of the olfactory adaptation paradigm, which they used to design a genetic screen to isolate mutants defective in their ability to adapt to volatile odors sensed by the Amphid Wing cells type C (AWC) sensory neurons. When wildtype C. elegans animals are exposed to an attractive AWC-sensed odor3 for 30 min they will adapt their responsiveness to the odor and will then ignore the adapting odor in a chemotaxis behavioral assay for ~1 hr. When wildtype C. elegans animals are exposed to an attractive AWC-sensed odor for ~1 hr they will then ignore the adapting odor in a chemotaxis behavioral assay for ~3 hr. These two phases of olfactory adaptation in C. elegans were described as short-term olfactory adaptation (induced after 30 min odor exposure), and long-term olfactory adaptation (induced after 60 min odor exposure). Later work from L'Etoile et al.,4 uncovered a Protein Kinase G (PKG) called EGL-4 that is required for both the short-term and long-term olfactory adaptation in AWC neurons. The EGL-4 protein contains a nuclear localization sequence that is necessary for long-term olfactory adaptation responses but dispensable for short-term olfactory adaptation responses in the AWC4. By tagging EGL-4 with a green fluorescent protein, it was possible to visualize the localization of EGL-4 in the AWC during prolonged odor exposure. Using this fully functional GFP-tagged EGL-4 (GFP::EGL-4) molecule we have been able to develop a molecular readout of long-term olfactory adaptation in the AWC5. Using this molecular readout of olfactory adaptation we have been able to perform both forward and reverse genetic screens to identify mutant animals that exhibit defective subcellular localization patterns of GFP::EGL-4 in the AWC6,7. Here we describe: 1) the construction of GFP::EGL-4 expressing animals; 2) the protocol for cultivation of animals for long-term odor-induced nuclear translocation assays; and 3) the scoring of the long-term odor-induced nuclear translocation event and recovery (re-sensitization) from the nuclear GFP::EGL-4 state.

A Molecular Readout of Long-term Olfactory Adaptation in C. elegans

Here we describe a molecular readout of long-term olfactory adaptation in Caenorhabditis elegans. The Protein Kinase G, EGL-4, is necessary for stable adaptation responses in the primary sensory neuron pair called AWC. During prolonged odor exposure EGL-4 translocates from the cytosol to nucleus of the AWC.

长期的嗅觉适应性的分子读数<em&gt; C.线虫</em

During sustained stimulation most sensory neurons will adapt their response by decreasing their sensitivity to the signal. The adaptation response helps ...

Direct Intraventricular Delivery of Drugs to the Rodent Central Nervous System

Due to an inability to cross the blood brain barrier, certain drugs need to be directly delivered into the central nervous system (CNS). Our lab focuses specifically on antisense oligonucleotides (ASOs), though the techniques shown in the video here can also be used to deliver a plethora of other drugs to the CNS. Antisense oligonucleotides (ASOs) have the capability to knockdown sequence-specific targets 1 as well as shift isoform ratios of specific genes 2. To achieve widespread gene knockdown or splicing in the CNS of mice, the ASOs can be delivered into the brain using two separate routes of administration, both of which we demonstrate in the video.
The first uses Alzet osmotic pumps, connected to a catheter that is surgically implanted into the lateral ventricle. This allows the ASOs to be continuously infused into the CNS for a designated period of time. The second involves a single bolus injection of a high concentration of ASO into the right lateral ventricle. Both methods use the mouse cerebral ventricular system to deliver the ASO to the entire brain and spinal cord, though depending on the needs of the study, one method may be preferred over the other.

We describe a method to target drugs to the central nervous system by either implanting a catheter or performing a bolus injection into the right lateral ventricle in mice. We focus specifically on the delivery of antisense oligonucleotides. This technique is readily adaptable to other drugs and to rats.

啮齿动物的中枢神经系统药物直接脑室交货

Due to an inability to cross the blood brain barrier, certain drugs need to be directly delivered into the central nervous system (CNS). Our lab focuses ...

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

The ability to probe defined neural circuits in awake, freely-moving animals with cell-type specificity, spatial precision, and high temporal resolution has been a long sought tool for neuroscientists in the systems-level search for the neural circuitry governing complex behavioral states. Optogenetics is a cutting-edge tool that is revolutionizing the field of neuroscience and represents one of the first systematic approaches to enable causal testing regarding the relation between neural signaling events and behavior. By combining optical and genetic approaches, neural signaling can be bi-directionally controlled through expression of light-sensitive ion channels (opsins) in mammalian cells. The current protocol describes delivery of specific wavelengths of light to opsin-expressing cells in deep brain structures of awake, freely-moving rodents for neural circuit modulation. Theoretical principles of light transmission as an experimental consideration are discussed in the context of performing in vivo optogenetic stimulation. The protocol details the design and construction of both simple and complex laser configurations and describes tethering strategies to permit simultaneous stimulation of multiple animals for high-throughput behavioral testing.

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

Optogenetics has become a powerful tool for use in behavioral neuroscience experiments. This protocol offers a step-by-step guide to the design and set-up of laser systems, and provides a full protocol for carrying out multiple and simultaneous in vivo optogenetic stimulations compatible with most rodent behavioral testing paradigms.

在啮齿动物中枢神经系统的体内光遗传学刺激

The ability to probe defined neural circuits in awake, freely-moving animals with cell-type specificity, spatial precision, and high temporal resolution ...

Isolation, Culture and Long-Term Maintenance of Primary Mesencephalic Dopaminergic Neurons From Embryonic Rodent Brains

Degeneration of mesencephalic dopaminergic (mesDA) neurons is the pathological hallmark of Parkinson&#8217;s diseae. Study of the biological processes involved in physiological functions and vulnerability and death of these neurons is imparative to understanding the underlying causes and unraveling the cure for this common neurodegenerative disorder. Primary cultures of mesDA neurons provide a tool for investigation of the molecular, biochemical and electrophysiological properties, in order to understand the development, long-term survival and degeneration of these neurons during the course of disease. Here we present a detailed method for the isolation, culturing and maintenance of midbrain dopaminergic neurons from E12.5 mouse (or E14.5 rat) embryos. Optimized cell culture conditions in this protocol result in presence of axonal and dendritic projections, synaptic connections and other neuronal morphological properties, which make the cultures suitable for study of the physiological, cell biological and molecular characteristics of this neuronal population.

The causes of degeneration of midbrain dopaminergic neurons during Parkinson&#8217;s disease are not fully understood. Cellular culture systems provide an essential tool for study of the neurophysiological properties of these neurons. Here we present an optimized protocol, which can be utilized for in vitro modeling of neurodegeneration.

原发性脑多巴胺能神经元的分离，培养和长期维持胚胎脑鼠类

Degeneration of mesencephalic dopaminergic (mesDA) neurons is the pathological hallmark of Parkinson&#8217;s diseae. Study of the biological processes ...

A Simplified Method for Ultra-Low Density, Long-Term Primary Hippocampal Neuron Culture

Culturing primary hippocampal neurons in vitro facilitates mechanistic interrogation of many aspects of neuronal development. Dissociated embryonic hippocampal neurons can often grow successfully on glass coverslips at high density under serum-free conditions, but low density cultures typically require a supply of trophic factors by co-culturing them with a glia feeder layer, preparation of which can be time-consuming and laborious. In addition, the presence of glia may confound interpretation of results and preclude studies on neuron-specific mechanisms. Here, a simplified method is presented for ultra-low density (~2,000 neurons/cm2), long-term (&#62;3 months) primary hippocampal neuron culture that is under serum free conditions and without glia cell support. Low density neurons are grown on poly-D-lysine coated coverslips, and flipped on high density neurons grown in a 24-well plate. Instead of using paraffin dots to create a space between the two neuronal layers, the experimenters can simply etch the plastic bottom of the well, on which the high density neurons reside, to create a microspace conducive to low density neuron growth. The co-culture can be easily maintained for &#62;3 months without significant loss of low density neurons, thus facilitating the morphological and physiological study of these neurons. To illustrate this successful culture condition, data are provided to show profuse synapse formation in low density cells after prolonged culture. This co-culture system also facilitates the survival of sparse individual neurons grown in islands of poly-D-lysine substrates and thus the formation of autaptic connections.

Low density cultures of primary hippocampal neurons usually require glia feeder layer to supply neurotrophic factors and sustain longevity. We describe here a simplified method to culture ultra-low density neurons on glass coverslips in the presence of a high density neuronal feeder layer, which facilitates investigation of specific neuronal-autonomous mechanisms.

的简化方法超低密度，长期主要海马神经元文化

Culturing primary hippocampal neurons in vitro facilitates mechanistic interrogation of many aspects of neuronal development. Dissociated embryonic ...

Analyzing Neural Activity and Connectivity Using Intracranial EEG Data with SPM Software

Measuring neural activity and connectivity associated with cognitive functions at high spatial and temporal resolutions is an important goal in cognitive neuroscience. Intracranial electroencephalography (EEG) can directly record electrical neural activity and has the unique potential to accomplish this goal. Traditionally, averaging analysis has been applied to analyze intracranial EEG data; however, several new techniques are available for depicting neural activity and intra- and inter-regional connectivity. Here, we introduce two analytical protocols we recently applied to analyze intracranial EEG data using the Statistical Parametric Mapping (SPM) software: time-frequency SPM analysis for neural activity and dynamic causal modeling of induced responses for intra- and inter-regional connectivity. We report our analysis of intracranial EEG data during the observation of faces as representative results. The results revealed that the inferior occipital gyrus (IOG) showed gamma-band activity at very early stages (110 ms) in response to faces, and both the IOG and amygdala showed rapid intra- and inter-regional connectivity using various types of oscillations. These analytical protocols have the potential to identify the neural mechanisms underlying cognitive functions with high spatial and temporal profiles.

We present two analytical protocols that can be used to analyze intracranial electroencephalography data using the Statistical Parametric Mapping (SPM) software: time-frequency statistical parametric mapping analysis for neural activity, and dynamic causal modeling of induced responses for intra- and inter-regional connectivity.

利用 SPM 软件分析颅内脑电图数据的神经活动和连接性

Measuring neural activity and connectivity associated with cognitive functions at high spatial and temporal resolutions is an important goal in cognitive ...

Long-term Sensory Conflict in Freely Behaving Mice

Long-term sensory conflict protocols are a valuable means of studying motor learning. The presented protocol produces a persistent sensory conflict for experiments aimed at studying long-term learning in mice. By permanently wearing a device fixed on their heads, mice are continuously exposed to a sensory mismatch between visual and vestibular inputs while freely moving in home cages. Therefore, this protocol readily enables the study of the visual system and multisensory interactions over an extended timeframe that would not be accessible otherwise. In addition to lowering the experimental costs of long-term sensory learning in naturally behaving mice, this approach accommodates the combination of in vivo and in vitro experiments. In the reported example, video-oculography is performed to quantify the vestibulo-ocular reflex (VOR) and optokinetic reflex (OKR) before and after learning. Mice exposed to this long-term sensory conflict between visual and vestibular inputs presented a strong VOR gain decrease but exhibited few OKR changes. Detailed steps of device assembly, animal care, and reflex measurements are hereby reported.

The presented protocol produces a persistent sensory conflict for experiments aimed at studying long-term learning. By permanently wearing a fixed device on their heads, mice are continuously exposed to a sensory mismatch between visual and vestibular inputs while freely moving in home cages.

自由生育小鼠的长期感觉冲突

Long-term sensory conflict protocols are a valuable means of studying motor learning. The presented protocol produces a persistent sensory conflict for ...

Assessment of Long-term Depression Induction in Adult Cerebellar Slices

Synaptic plasticity provides a mechanism for learning and memory. For cerebellar motor learning, long-term depression (LTD) of synaptic transmissions from parallel fibers (PF) to Purkinje cells (PC) is considered the basis for motor learning, and deficiencies of both LTD and motor learning are observed in various gene-manipulated animals. Common motor learning sets, such as adaptation of the optokinetic reflex (OKR), the vestibular-ocular reflex (VOR), and rotarod test were used for evaluation of motor learning ability. However, results obtained from the GluA2-carboxy terminus modified knock-in mice demonstrated normal adaptation of the VOR and the OKR, despite lacking PF-LTD. In that report, induction of LTD was only attempted using one type of stimulation protocol at room temperature. Thus, conditions to induce cerebellar LTD were explored in the same knock-in mutants using various protocols at near physiological temperature. Finally, we found stimulation protocols, by which LTD could be induced in these gene-manipulated mice. In this study, a set of protocols are proposed to evaluate LTD-induction, which will more accurately allow examination of the causal relationship between LTD and motor learning. In conclusion, experimental conditions are crucial when evaluating LTD in gene-manipulated mice.

In some gene-manipulated animals, using a single protocol may fail to induce LTD in cerebellar Purkinje cells, and there may be a discrepancy between LTD and motor learning. Multiple protocols are necessary to assess LTD-induction in gene-manipulated animals. Standard protocols are shown.

成人脑切片长期抑郁症诱导评估

Synaptic plasticity provides a mechanism for learning and memory. For cerebellar motor learning, long-term depression (LTD) of synaptic transmissions from ...