Name: vagus nerve stimulation as a tool to induce plasticity in pathways relevant for extinction learning
Uploaded: 2015-08-21T14:00:00.000+00:00
Duration: 11 min 2 s
Description: Watch this Scientific Journal Video about Vagus Nerve Stimulation as a Tool to Induce Plasticity in Pathways Relevant for Extinction Learning at JoVE.com

This research was supported by the National Institute of Mental Health MH 086960-01A1 (Christa K. McIntyre).

在这个协议中描述的所有程序进行，根据美国国立卫生研究院指南实验动物的护理和使用，他们被批准由得克萨斯大学达拉斯分校的机构动物护理和使用委员会。 1.建设VNS袖口<ol><li>创建一个钻井工具被锯掉一个22½摹针的尖底。 </li><li>运行22个半摹针现在钝端了一个金属文件多次压平。握住针以45度角的文件并运行它多次在该文件同时旋转它。这将导致金属变得更薄和卷起向内。注意:插入手术刀的尖端插入针的切割端，并与一些向下的力转动到打出的金属。 </li><li>用放大镜望远镜在第1节的其余步骤。 </li><li>使用手术刀（10或15刀片）切割管材为4mm段。 </li><li>将一个4毫米段过小钻头或其他类似形状的工具。这是为了保持管件的地方，当它被操纵。 </li><li>钻4个孔入管（ 图1A）。孔应通过2mm见方使四个点的2毫米和应清洁，无粗糙的边缘。 注:该钻井工具需要被重磨（步骤1.2），每2或3个孔。难度与这个步骤几乎可以肯定是由于钻井工具不够清晰。 </li><li>与油管仍然在钻头，使用手术刀沿纵向切开的孔之间的管道，使得两个孔最终在切割（ 图1B）的任一侧。 </li><li>用缝纫针和缝合线，通过缝合穿过孔以创建索具周围的迷走神经的压脉袋的最后的位置。开始与袖带内的针和通过孔之一通它，然后再返回经过从（ 图1C）的外侧相邻的孔。扳平thread从塑料一起〜2厘米使得管和螺纹作出一个三角形。允许〜8厘米线结后修剪。重复此过程，在切割的相对侧上的孔。该管道是现在可以连线。 </li><li>准备电线袖口。 <ol><li>削减铂铱丝成70毫米段。 </li><li>使用最好的小费珠宝火炬，创造一个尖锐的火焰，蓝色中心，这是因为精炼越好，使用它剥去约1厘米的电线塑料涂层。适用于火焰的蓝色中心到导线的裸端创建一个小球。应用火焰的蓝色中心到几个百分点的剥离部分的熔合七股线一起。在这些点线将出现扭结。 </li><li>在电线上的相对端，施加火焰的蓝色中心到端创建一个小球。最小化剥离塑料就这样结束了。 </li></ol></li><li>接线箍（ 图1D）。 注:科技教育PS在第1.10必须在放大镜望远镜来完成。 <ol><li>用胶带将袖口向下使用线程，以便它的方向为切割运行水平。紧拉的螺纹，使得袖带拉开，然后用胶带向下的螺纹。 </li><li>抓住电线准备用5＃镊子剥离一侧的球状端和推动通过右下角的孔。拉钳子出洞，而使导线的端部，在压脉袋的中间。 </li><li>重新把握裸线的球状端部（现在箍的中间），并通过右上方孔推。拉钳子出洞，而使通过在压脉袋的顶侧的箍和松散完全通过电线。 </li><li>重新把握裸线的球状端部（现在在顶侧袖口外），并将其推回通过从内侧右上方孔。继续这样做，直到电线固定到位:通过测试拉着线的另一端​​。 注意:在此过程中是非常关键的区分电线的剥离/绝缘部分。最终将被定位在压脉袋的"槽"的金属丝必须被剥离，但底部孔内（底端压脉袋外）下的所有内容必须绝缘。这确保输送电流只向迷走神经。 </li><li>把握绝缘侧的球状端，并通过从所述压脉袋的内部右下方孔推，循环它周围一次。 </li><li>重复步骤1.10.2 - 1.10.5的袖带，使两根导线最终固定到管道，一个在右侧，另一个在左侧的左侧。 </li><li>将金销入伸出援手与孔朝上的手臂。填充焊剂的孔。 </li><li>焊接导线（现在连接于袖带）插入针的绝缘端。 </li><li>允许焊料冷却，然后再次熔化。这确保了导线的端部和i之间的良好的连接nside引脚。如有必要，应用更焊料。重复第二线。 </li><li>与运行在正确的引线，标记用永久性标记箍的顶部。还标记附着在顶端引线的金销。 </li></ol></li></ol> 2.施工Headcap的VNS输入网站<ol><li>削减26 AWG铜线30毫米段。剥去端的一小部分。 </li><li>切窄端关闭松金销和焊接线的剥离端的金脚的切割端。创建两个线/针化合物为每个所需的输入位点。放置一个连接器插入帮助手和焊接的线/针化合物与每个连接器（ 图2G）的两个助熔齿的线端。 </li><li>标记与Sharpie笔的电线中的一个。在手术过程中，定位与所述标记的导线喙无标记线的植入物。 </li></ol> 3.手术VNS <ol><li>搬运vagu的创建自定义工具玻璃手术在S神经。 <ol><li>使用硼硅酸盐玻璃拉微量，使其具有长的锥形尖端。如果没有吸管拉马可用，打破玻璃打造一个较长的边缘。 </li><li>持用厚布的移液管的非锥形端（防止烫伤），然后按锥形/断端的成光滑耐火表面而施加的蓝色火焰从珠宝炬到锥形端。因为它被压入表面玻璃会弯曲。用明火直到钩或J形形式。 </li></ol></li><li> VNS手术过程<ol><li>收集和消毒所有的工具。准备一个卫生，加热手术区。 </li><li>麻醉动物用氯胺酮/赛拉嗪（85毫克/公斤，5毫克/公斤，IP）。通过监测动物的叫声和戒断反射响应于脚趾和/或尾巴捏评估麻醉平面的深度。 </li><li>剃头动物的脖子的顶部和左侧。保护动物的眼睛矿工人油或眼膏。申请用纱布清洁碘溶液，然后用酒精纱布剃光领域。重复一次。 </li><li>注入0.05毫升麻卡因皮下上的头顶部，并允许丸药以同时将动物放入立体定位仪分散。 </li><li>用手术刀做一个切口在头骨上的皮肤都暴露Lambda和前囟门。通过使用钝钳到隧道皮下从切口部位向下的左侧耳前到颈部左侧准备袖带的路径。 </li><li>拉切口部位止血与开放。使用棉签，应用过氧化氢，以暴露颅骨以除去任何剩余的组织。 </li><li>使用手术刀，钻两个浅浅的首发洞的头骨放置锚螺丝。它们应该放置得足够远，以允许室的植入物，但不能太靠近周围的组织。避免直接对中线放置螺丝。 <ol><li>随着力ps和螺丝刀，带动骨螺钉到这两个孔。螺钉应紧在孔，与帽2 - 3毫米头骨表面上方以允许余地丙烯酸填写下方和周围的螺丝。 </li></ol></li><li>下填充空间，左右之间，以及用少量的丙烯酸的螺钉，避免周围组织。然后将丙烯酸的用量较大，​​在两个螺钉之间的颅骨的中间。 </li><li>抓住植入物，以便标记线被定向喙无标记线和快速地放置植入物中的丙烯酸类，注意不要让丙烯酸入金销，卡环区，或在上面的输入位点。一旦定位允许设置〜5分钟，待干。这也可以使用立体定位为支撑的臂实现。在植入和颅骨与丙烯酸的低粘度的混合之间的任何裂缝或缺口填补。晾干。 </li><li>用放大镜望远镜在3.2节中的其余步骤。 </li><li>第删除Ë动物从立体定位仪。放置在其右侧的动物，稍微转动而朝向腹侧位置。 </li><li>使大约在左颈静脉小切口。颚骨和锁骨应大致等距离到切口部位。使用钝器解剖扩大切口，直至到达肌肉层。该胸锁乳突肌，胸骨舌骨肌，肩胛舌骨和肌肉应该是可见的。用拉钩肌肉保持网站开放。 </li><li>继续钝解剖沿着肌肉之间的自然沟。寻找颈动脉的脉动。通过向脉冲肌肉标题将揭示颈动脉。拉背部肌肉与肌肉的牵引器。含有颈动脉鞘也含有迷走神经。仔细解剖钝的剪刀护套。 </li><li>确定迷走神经。它是在颈动脉鞘最大神经，通常是对动脉的动物的左侧但可以在任何一侧找到。切换到定制玻璃工具和从颈动脉分开的迷走神经。神经应无任何其它组织的至少5毫米。 </li><li>从切口上的头，使用该螺纹上的引线相对的压脉袋的一侧，通过预先制成皮下隧道用镊子或小止血拉袖带。通过组织将其推入切口部位在颈部。 </li><li>使用玻璃工具轻轻抬起神经向上推的螺纹上的神经下引线相对的压脉袋的一侧。一路经过拉线程，注意不要划到神经。袖带应立即相邻的神经。 </li><li>确保袖口的取向是标有&#39;顶&#39;端优越。拖放神经入箍的中间。神经现在应该跨两个导线在于袖带（ 图2B）的槽。绑在一起的线程收袖口。 </li><li>在headcap，插上连接到袖口进入黄金引脚上的刺激输入网站的引脚。请将标销插入附着在模拟输入网站最前牙的金脚。 </li><li>以验证该箍正确刺激迷走神经，执行呼吸测试通过，所述刺激器连接到刺激输入网站上headcap和运行刺激的停止（0.3毫安，60赫兹，最多10秒）。呼吸要短暂停留和心脏率应该放弃，袖带验证功能。 </li><li>安全的刺激输入网站与丙烯酸销。覆盖电线，并验证暴露针和线不会导致短路。使用丙烯酸平滑过度颠簸的任何或填写任何空白。晾干。 </li><li>关闭缝合切口两个站点。注入0.05毫升麻卡因皮下接近颈部切口部位。应用抗生素软膏切口部位。任选地，离开阳连接器就位在刺激输入位置到防止在愈合过程中的损坏或阻塞。 </li><li>对待抗生素，并按照标准术后护理包括适当的镇痛。回到动物畜舍设施的恢复，他们的流动性后。允许恢复5天。为了保证最大的长寿和headcap功能，房子单独的动物为实验的其余部分。 注:假手术VNS大鼠经历相同的手术，但该电路被设计成短于headcap的水平（ 即，一个headcap植入和迷走神经是从颈动脉中分离，但没有电极套周围放置神经）。 </li></ol></li></ol> 4.听觉恐惧条件注:此恐惧条件协议较为密集比大多数21，因为这些实验的目标是提高灭绝。用温和的恐惧条件，很容易熄灭，地面效应可以掩盖这一增强。 </p&gt; <ol><li>房子的动物在12小时的光/暗循环随意获得食物和水。从手术后恢复期间每天处理的动物。 </li><li>建立调节和检测仪器，包括一个操作性箱装在一个声音衰减室（ 图2C）的。操作性盒具有透明塑料壁，20×20×20厘米，并有一个不锈钢格栅地板，它连接到一个足电击发生器。使用白色的房子光照亮室视频录制。使用9千赫，85分贝声压级音作为条件刺激。 </li><li>使用位于腔室内，上述的操作性框的数字照相机记录的行为。查看和监控位于客房行为以外的计算机上的会话。保存视频供日后分析。 </li><li>擦拭用70％乙醇室前，每次会议之后，以消除嗅觉线索。 </li><li>恐惧条件大鼠2天（ 图3A）。确认老鼠是不纳利怕口气由第一天呈现5吨（9千赫，85分贝，30秒）。确保冻结水平是可以忽略不计。 <ol><li>按照最初的基调演讲有8音电击（1秒，0.5 mA）的配对在每个连续2天。第二天再重复音电击配对。变化2和4分钟之间的相互刺激间隔（ISI），平均3分钟，每试验。随机在该震荡基调期间发生点。 </li></ol></li><li>第三天，测试音/休克协会的力量。播放4的色调与3在没有footshocks的ISI，4或5分钟（4分钟平均值），并且在音调介绍和在刺激间间隔中作为条件恐惧响应措施记录动物的冷冻行为（CFR）。 </li><li>第4天，开始消退训练与迷走神经刺激或深水VNS。 <ol><li>从stimulato将阳连接器将老鼠变成了刺激r记录到刺激输入位置。地方动物进入所述腔室（ 图2A，2C）。在30赫兹设置刺激〜0.4毫安，500微秒的脉冲宽度。设置刺激的30.15秒的总时间，开始150毫秒的基调发病前。扮动物4种色调（如步骤4.6）和配对每一个音调呈现与迷走神经刺激或深水VNS。 </li></ol></li><li>定期测试的袖口和投入现场使用示波器的电气完整性。 <ol><li>堵塞动物在正常和从刺激到示波器分割的输出。 </li><li>设置示波器为-20 V范围内，以+ 20V和运行刺激。在30赫兹的刺激波形应该是在示波器上可见。超刺激的大小10 V表示高阻状态和不当运作袖口或刺激输​​入站点连接。 </li></ol></li><li>上测试消退训练VNS的效果运行第二CFR测试（如在步骤4.6）在第5天记录所花的时间中陈述语气冻结和比较第一CFR测试（步骤4.6）中记录基线冻结。 </li><li>分析采用独立的观察员谁是盲目的治疗条件的视频。测量时间花费在用秒表基调演讲冻结。冻结被定义为完整不动，在此期间，大鼠表现出快速呼吸，降低头部，并扩散爪子22。冻结行为分析，可以分为两个阶段:在基调演讲和刺激间的时间间隔中。 </li></ol> 5. 在诱发电位的体内录音注意:此步骤是可选的。诱发电位（交易所指数基金）的异氟烷麻醉大鼠安装在立体定向仪，按照标准程序23,24被记录复职的测试后24小时（5天）。 <ol><li>收集和消毒所有的工具。 </li><li>诱导麻醉用异氟烷（5％在100％的氧气，流量1升/分钟）在一个透明的塑料腔室。通过监测动物的叫声和戒断反射响应于脚趾和/或尾巴捏评估麻醉平面的深度。使用矿物油或眼药膏，以保护眼睛。 </li><li>注入0.05毫升麻卡因皮下上的头顶部，并允许快速浓分散。用手术刀和止血从头骨取出headcap。用尽可能少的力量，尽可能避免混淆前囟门。 </li><li>将动物在立体定位仪。用手术刀扩大切口，显露两个拉姆达和前囟门。维持麻醉平面经由鼻锥异氟烷（在100％的氧气3％，流量1升/分钟）。 </li><li>钻孔进入infralimbic前额叶皮质（IL）和基底外侧杏仁核（BLA）以上的头骨。低的玻璃微电极（2M氯化钾1-2兆欧电阻）到BLA（D / V:7.2，A / P:2.7，M / L:4.9，从囟）和stimulat离子电极插入内侧前额叶皮质的IL区（D / V:4.6，A / P:3.0，M / L:0.7，从囟）（ 图4A）。 </li><li>刺激IL唤起了BLA期货转现货。在图4中所示的数据是使用以下设置获得:0.3毫秒持续时间阿刺激脉冲，使用刺激强度是对应于最小电流强度是诱发的最大场响应的40％（基于之前确定的输入输出曲线上收集基线数据），每交付15秒。 </li><li>诱导突触可塑性前15分钟 - 收集了至少10基准数据。 注意:用于唤起突触可塑性将随实验的要求，并且必须由每个实验者仔细选择的协议。在图4C的数据示出了在最小电流强度，唤起变化在以下3个脉冲串的100个脉冲，在50赫兹（2秒）的EFP，用20秒间脉冲间隔d是最大场响应。 </li><li>测量EFP的振幅作为所述刺激伪迹之前和5毫秒窗口的平均值之间的差的5毫秒窗口约20的平均值 - 25毫秒的刺激伪迹后，对应于该场电位的负峰值。正常化数据到基线，并设置一个10分钟基线的平均为100％。使用的另外10分钟期间的平均EFP振幅可塑性诱导后（ 例如，40 - 50分钟后诱导），以评估在EFP振幅的长期变化。 </li><li>在记录结束后斩首麻醉动物并提取大脑。准备组织进行电极放置组织学检验。 </li></ol>

The authors have no competing interests or conflicts.

我们在这里是，用于曝光的单个会话条件线索19期间促进的条件性恐惧灭绝和调节可塑性在infralimbic皮质和基底外侧杏仁核可能介导消光学20之间的通路的协议。此协议的成功的一个关键步骤是VNS的过程中消退训练正确的交付。因此，应特别注意给予袖带电极的构造和安置周围迷走神经的袖口。在袖带电极的施工过程中，它确保导线的暴露部分是在正确的位置是很重要的。同样地，在手术过程中，应特别努力放置在适当的位置上的压脉袋，并以使压脉袋和所述刺激器之间的电路保持完好在实验的过程中充分地锚定在适当的位置。的压脉袋的正常功能应该b手术后E经过行为测试后（如在步骤3.2.20所述），并再次（如在步骤4.8中概述）。 通过该VNS调节活性在中枢神经系统中的机制尚未完全了解。 ，颈迷走神经是由传入感觉和传出运动纤维在约4比1的比例分别为25。迷走神经传入中继信号到孤束核（NTS），然后项目臂旁核，下丘脑，丘脑，杏仁核，海马和26,27。重要的是，一元胺原子核在脑干，蓝斑（LC）和所述缝核，接收来自NTS直接和/或间接突起（ 图5）。从而VNS可能经由多个神经调节的协同作用调节皮质可塑性和记忆。有关迷走神经刺激提出的影响包括改变去甲肾上腺素（NE）的释放通过从NTS到突出LC，抑制GABA水平升高，并通过网状激活系统抑制28,29,30异常皮层活动。乙酰胆碱，血清素，脑源性神经营养因子的重要作用也被证明31-35。对恐惧记忆的调制和消光一般，迷走神经刺激对发射机释放在前额皮质的影响（PFC），杏仁核和海马很可能是特别相关36,37。急性VNS增加去甲肾上腺素和血清素的释放同时在内侧PFC 33,38和杏仁核39,30。急性VNS也增加去甲肾上腺素38的水平，并增强突触传递海马40-42。去甲肾上腺素以前已经显示参与恐惧表达的调节。从LC到前脑的NE预测病变损害主动回避的灭绝不改变收购原稿或保留INAL学习43,44。的条件性恐惧固结取决于β受体激活BLA 43内，并且一些报告建议为α-和β肾上腺素受体 ​​在内侧的PFC两个毒品和恐惧消退训练46-49的记忆巩固的作用。因此，随着迷走神经刺激，迷走神经刺激诱发的神经调节的释放像元或5-HT配对消退训练时出现以促进突触可塑性而导致的训练单独，导致灭绝的增强合并。 VNS的主要优势在于其时空特异性。与全身或局部甚至药物的应用，这往往有一个缓慢的发病和行动失调，VNS可以选择搭配特定的行为来促进这些调节利益的行为主动网络突触可塑性。我们在这里描述使用的参数也可用于临床的treatme一个VNS协议癫痫在人类新台币（0.6毫安，500微秒脉冲宽度为30赫兹，30秒上5分钟关闭的刺激循环）。微透析实验表明，采用迷走神经刺激这些参数会导致网元释放的杏仁核39一个持久的，大致2倍的增加。这个大而相对缓慢增加NE可以解释为什么VNS搭配消退训练，不像扩展消退训练本身，也促进冻结条件刺激的呈现（在intertrial间隔）以外的行为的灭绝。因为在我们的实验动物，在相同的上下文中听觉恐惧条件进行消退训练，在ITI过程中冻结行为的减少表明，VNS促进灭绝学习推广到上下文。 然而，尽管对NE释放潜在的长期持久的影响，VNS对行为和突触可塑性的影响，只有当他们配对W¯¯明显第i个特定的行为。迷走神经刺激后不久恐惧消退训练大鼠在其homecages应用没有促进消光20，并且类似地，迷走神经刺激施加一个特定行为上下文之外没有改变的IL-BLA途径（参见图3C）的突触可塑性。另一方面，即使是短暂VNS的应用（ 例如 ，0.8毫安作为15个脉冲，在30赫兹的500毫秒100微秒脉冲宽度的火车）引起感觉或运动网络特定和持久的变化，当它们被输送然与正在进行的行为。 迈克尔Kilgard的组7的开创性论文表明，VNS的时间精确配对的单一色调的演示文稿导致可塑性音调频率的听觉皮层的地图。这些影响已经用于临床逆转病理可塑性听觉皮层来治疗耳鸣50。同样，迷走神经刺激的重复配对与马达行为已经显示重组运动皮层8，这靶向可塑性是在功能在行程9,10的不同的动物模型中的恢复非常有效。 然而，延迟VNS康复训练或VNS的几倍较大数额交付后2小时导致比精确定时VNS 51的改进相对较少。因此，未来的研究需要优化的时间安排和迷走神经刺激，以最大限度地提高治疗益处的量。我们的研究结果表明迷走神经刺激，这是临床上批准用于多药耐药性癫痫和难治性抑郁症的治疗中，可以使用作为辅助治疗，以暴露疗法，因为它调制学习特定可塑性以提高暴露对消光的效果条件性恐惧反应。

古典恐惧条件提供了一种广泛使用的动物模型来研究焦虑症的生物学基础。期间恐惧条件，厌恶的刺激（非条件刺激，US，例如 ，一个电击）的输出，结合一个中性刺激，例如铃声和/或上下文（条件刺激; CS）。在恐惧条件，CS和美国之间的关联而形成的。最终，CS的单独呈现一个引起恐惧反应（该条件反应; CR）。在恐惧消退中，CS反复提出在没有美国，造成CR到逐渐减少1。因此，条件性恐惧灭绝是其中可怕的行为反应，以中性刺激衰减，当他们不再预测厌恶成果的活动进程。是不是条件反射灭绝，需要巩固与协会了解到新的竞争回忆。焦虑症的特点是IMPAIRED灭绝2-4。因此，条件性恐惧的动物模型灭绝同时作为抑制性的学习和行为作为治疗人类焦虑症5,6模型的一个重要范例。 因为恐惧和焦虑的人之间的紧密对应关系，它认为这些研究可以提供洞察焦虑有关的疾病如创伤后应激障碍的生物特性，将帮助制定战略对待他们。临床前研究的一个重要目标是帮助灭绝学习和诱导针对性可塑性灭绝电路，以巩固学习灭绝。迷走神经刺激（VNS）是可能被用于提供的脑区和接合在一个正在进行的任务突触紧时间和电路的特定调制微创神经假方法。最近的一系列研究，从迈克尔Kilgard小组在得克萨斯大学达拉斯分校有表明配对VNS与分立感觉或运动的刺激（ 如铃声或杆拉）是促进大脑皮层的可塑性治疗耳鸣7，或中风后8-10克服运动障碍非常有效。此外，非偶然VNS发生在很短的时间窗口后，学习同样促进大脑皮层的可塑性，增强记忆的巩固大鼠和人类中11-13。 考虑在副交感神经通路的迷走神经的作用，这并不奇怪，它可能参与调节记忆和突触可塑性。高度情绪化的事件往往产生于非情感记忆更强的回忆。这可能是由于应激激素对记忆巩固的影响。应激激素肾上腺素Posttraining施用增强记忆巩固在人类和非人类动物，但肾上腺素不穿过血-脑屏障14,15 </suP&gt;。因此，压力引起的肾上腺素释放，必须影响大脑间接地增强记忆的巩固。有力的证据表明，在迷走神经可以是循环肾上腺素和大脑之间的联系。宫下和威廉姆斯16发现，肾上腺素的全身给药提高迷走神经发射，并增加去甲肾上腺素水平的杏仁核17。肾上腺素全身用药不增强记忆的巩固，当β肾上腺素能受体阻断杏仁核18表明迷走神经在，变成情感激发经验转化为长期记忆通路的作用。 因此，VNS配对与培训，以提高支持记忆的巩固和暴露于空调线索在没有加固的增强灭绝学习大鼠19,20巩固脑部变化的潜力。在这里，我们描述了使用的一个VNSSA工具，以促进大脑皮层的可塑性和便利条件性恐惧反应灭绝。

<table><tbody><tr><td>Alcohol</td><td></td><td></td><td></td></tr><tr><td>Atropine</td><td>Fisher</td><td>A0132-5G</td><td></td></tr><tr><td>Betadine</td><td>Henry Schein</td><td>69066950</td><td></td></tr><tr><td>Hydrogen peroxide&amp;nbsp;</td><td>CVS</td><td>209478</td><td></td></tr><tr><td>Ketamine</td><td>Henry Schein&amp;nbsp;</td><td>1129300</td><td></td></tr><tr><td>Marcaine</td><td>Henry Schein</td><td>6312615</td><td></td></tr><tr><td>Mineral Oil</td><td>CVS</td><td>152355</td><td></td></tr><tr><td>Neosporin</td><td>CVS</td><td>629451</td><td></td></tr><tr><td>Oxygen</td><td>Home Depot</td><td>304179</td><td></td></tr><tr><td>Pennicillin</td><td>Fisher</td><td>PENNA-10MU</td><td></td></tr><tr><td>Propane</td><td>Home Depot</td><td>304182</td><td></td></tr><tr><td>Xylazine</td><td>Henry Schein</td><td>4019308</td><td></td></tr><tr><td>Tools</td><td></td><td></td><td></td></tr><tr><td>Jewelery Torch</td><td>Smith Equipment</td><td>23-1001D</td><td></td></tr><tr><td>Sewing Needle</td><td>Walgreens</td><td>441831</td><td></td></tr><tr><td>#5 Forceps (2)</td><td>Fine Science Tools</td><td>11254-20</td><td></td></tr><tr><td>Soldering Iron</td><td>Home Depot&amp;nbsp;</td><td>203525863</td><td></td></tr><tr><td>AmScope SM-4TX-144A 3.5X-45X Circuit Board Boom Stereo Microscope + 144 LED</td><td>AmScope</td><td>SM-4TX-144A</td><td></td></tr><tr><td>Helping Hands</td><td>A-M Systems&amp;nbsp;</td><td>726200</td><td></td></tr><tr><td>Scalpel Blade Holder</td><td>Fine Science Tools</td><td>10003-12</td><td></td></tr><tr><td>Metal File</td><td>Home Depot</td><td>6601</td><td></td></tr><tr><td>Ruler</td><td>Home Deopt</td><td>202035324</td><td></td></tr><tr><td>Curved Hemostats&amp;nbsp;</td><td>Fine Science Tools</td><td>130009-12</td><td></td></tr><tr><td>Fine Scissors</td><td>Fine Science Tools</td><td>14058-09</td><td></td></tr><tr><td>Spatula</td><td>Fine Science Tools</td><td></td><td></td></tr><tr><td>Small Screwdriver</td><td>Home Depot</td><td>646507</td><td></td></tr><tr><td>Magnetic Fixator Retraction System</td><td>Fine Science Tools</td><td>18200-04, 18200-01, 18200-05</td><td></td></tr><tr><td>Heating Pad</td><td>Walgreens</td><td>30294</td><td></td></tr><tr><td>Clippers</td><td>Walgreens</td><td>277966</td><td></td></tr><tr><td>Sharpie</td><td>Staples</td><td>125328</td><td></td></tr><tr><td>Ring Forceps</td><td>Fine Science Tools</td><td>11103-09</td><td></td></tr><tr><td>Custom Micropipette Glass Tools (J shape and Straight) - Borosilicate glass</td><td>Sutter Instrument</td><td>B150-110-10</td><td></td></tr><tr><td>Adson Forceps</td><td>Fine Science Tools</td><td>11006-12</td><td></td></tr><tr><td>Cuffs</td><td></td><td></td><td></td></tr><tr><td>Tubing</td><td>Braintree Scientific Inc</td><td>MRE-065</td><td></td></tr><tr><td>Platinum Iridium Wire</td><td>Medwire</td><td>10IR9/49T</td><td></td></tr><tr><td>Gold Pins</td><td>Mill-Max</td><td>1001-0-15-15-30-27-04-0</td><td></td></tr><tr><td>Suture Thread</td><td>Henry Schein</td><td>100-5797</td><td></td></tr><tr><td>22 G needles</td><td>Fisher&amp;nbsp;</td><td>14-815-525</td><td></td></tr><tr><td>Paper Tape</td><td>Fisher&amp;nbsp;</td><td>03-411-602</td><td></td></tr><tr><td>Solder</td><td>Home Depot</td><td>327793</td><td></td></tr><tr><td>Flux&amp;nbsp;</td><td>Home Depot</td><td>300142</td><td></td></tr><tr><td>Scalpel Blade, 10 or 15</td><td>Stoelting</td><td>52173-10</td><td></td></tr><tr><td>Silastic Laboratory Tubing .51 mm ID x .94 mm OD</td><td>Fisher&amp;nbsp;</td><td>508-002</td><td></td></tr><tr><td>Headcaps</td><td></td><td></td><td></td></tr><tr><td>Connector Pieces (male)</td><td>Omnetics Connector Corporation</td><td>A25001-004</td><td></td></tr><tr><td>Headcap pieces (female)</td><td>Omnetics Connector Corporation</td><td>A24001-004</td><td></td></tr><tr><td>Teets Dental Acrylic, Liquid and Powder</td><td>A-M Systems</td><td>525000, 526000</td><td></td></tr><tr><td>26 Gauge Solid Copper Wire</td><td>Staples</td><td>1016882 &amp;nbsp;</td><td></td></tr><tr><td>Surgery</td><td></td><td></td><td></td></tr><tr><td>Bone Screws</td><td>Stoelting+CB33:C61</td><td>51457</td><td></td></tr><tr><td>Scalpel Blades, 10 or 15</td><td>Stoelting</td><td>52173-10</td><td></td></tr><tr><td>1 ml syringes</td><td>Fisher</td><td>14-826-261</td><td></td></tr><tr><td>22 G Needles</td><td>Fisher&amp;nbsp;</td><td>14-815-525</td><td></td></tr><tr><td>27 G Needles</td><td>Fisher</td><td>14-826-48</td><td></td></tr><tr><td>2&quot; x 2&quot; Gauze</td><td>Fisher</td><td>22-362-178</td><td></td></tr><tr><td>Swabs</td><td>Fisher</td><td>19-120-472</td><td></td></tr><tr><td>Puppy Pads</td><td>PetCo</td><td>1310747</td><td></td></tr><tr><td>Kim Wipes</td><td>Fisher</td><td>06-666-A</td><td></td></tr><tr><td>Chamber and Behavioral Setting&amp;nbsp;</td><td></td><td></td><td></td></tr><tr><td>Husky Metal Front Base Cabinet (30WX19DX34H)</td><td>Home Depot</td><td>100607961</td><td></td></tr><tr><td>Quiet Barrier&amp;shy; HD Soundproofing Material (Sheet) (PSA)</td><td>soundproofcow.com</td><td>10203041</td><td></td></tr><tr><td>Convoluted Acoustic Foam Panel</td><td>soundproofcow.com</td><td>10432400</td><td></td></tr><tr><td>Isolated Pulse Stimulator Model 2100</td><td>A-M Systems</td><td>720000</td><td></td></tr><tr><td>Digital Camera - Logitech Webcam C210</td><td>Logitech</td><td>B003LVZO88</td><td></td></tr><tr><td>MatLab</td><td>Mathworks.com</td><td></td><td></td></tr><tr><td>Sinometer 10 MHz Single Channel Oscilloscope</td><td>Sinometer</td><td>CQ5010C</td><td></td></tr><tr><td>OxyLED T-01 DIY Stick-on Anywhere 4-LED Touch Tap Light</td><td>OXYLED</td><td>B00GD8OKY0</td><td></td></tr><tr><td>5k ohm potentiomter</td><td>Alpha Electronics</td><td>B00CTWDHIO</td><td></td></tr><tr><td>Extech 407730 40-to-130-Decibel Digital Sound Level Meter</td><td>Extech Instruments</td><td>B000EWY67W</td><td></td></tr><tr><td>DSCK-C Dual Output, scrambled shocker</td><td>Kinder Scientific Co</td><td></td><td></td></tr></tbody></table>

vagus nerve stimulation as a tool to induce plasticity in pathways relevant for extinction learning

Extinction describes the process of attenuating behavioral responses to neutral stimuli when they no longer provide the reinforcement that has been maintaining the behavior. There is close correspondence between fear and human anxiety, and therefore studies of extinction learning might provide insight into the biological nature of anxiety-related disorders such as post-traumatic stress disorder, and they might help to develop strategies to treat them. Preclinical research aims to aid extinction learning and to induce targeted plasticity in extinction circuits to consolidate the newly formed memory. Vagus nerve stimulation (VNS) is a powerful approach that provides tight temporal and circuit-specific release of neurotransmitters, resulting in modulation of neuronal networks engaged in an ongoing task. VNS enhances memory consolidation in both rats and humans, and pairing VNS with exposure to conditioned cues enhances the consolidation of extinction learning in rats. Here, we provide a detailed protocol for the preparation of custom-made parts and the surgical procedures required for VNS in rats. Using this protocol we show how VNS can facilitate the extinction of conditioned fear responses in an auditory fear conditioning task. In addition, we provide evidence that VNS modulates synaptic plasticity in the pathway between the infralimbic (IL) medial prefrontal cortex and the basolateral complex of the amygdala (BLA), which is involved in the expression and modulation of extinction memory.

本节中示出了可以通过使用VNS结合灭绝学习，以减少在大鼠条件恐惧反应的表达来获得的结果的例子。对于1和2天（听觉恐惧条件反射），大鼠进行了培训，对听觉恐惧条件任务中footshocks进行搭配的基调。 3（预处理试验）日，音被提出在没有footshocks测量冰点水平，并推断出条件性恐惧反应的收购。第4天收到的（治疗）大鼠组特定消退训练和治疗​​:4基调演讲中搭配无论是VNS或假刺激或扩展灭绝组中，20基调演讲与假刺激。冷冻水平响应于4展示在CS单独（后处理试验）（ 图3A中比照时间轴）的再次测试的第5天。接收有限量的非增强曝光数（4），以共同的动物在消光相位nditioned色调只显示一个小的降低条件性恐惧以下复原天（ 图3B）。与此相反，VNS治疗的大鼠表现出显著减少在单个消退训练会话（ 图3B）后冷冻。减少VNS处理的动物的条件恐惧反应的量相当于中所见的，在消光训练（组EE在图3B）接收的非增强暴露（20吨）的5倍的量假处理的动物。使用以上概述的具体实验装置，迷走神经刺激也可以促进消到上下文。 如图3C所示 ，迷走神经刺激的动物还显示减少冷冻行为的调节色调的呈现之外，这表明它们的消退训练还推广到上下文。我们还表明，配对与迷走神经刺激消退训练改变的途径betwe的metaplasticity烯的infralimbic皮质素（IL）和基底外侧杏仁核（BLA）麻醉动物（ 图4）。在恐惧空调的动物，没有经过培训的灭绝，对当地诱发的领域在BLA的IL诱导LTD短暂爆发刺激（HFS）（ 图4）。无论他们一个灭绝会话期间给予延长灭绝培训或VNS，该突触抑郁症的表现动物的条件恐惧显著灭绝得到扭转;然而，迷走神经刺激的消失期间施用促进诱导LTP的同时动物扩展消光组中显示出响应的HFS没有变化。此结果中也观察到了被假刺激了四色调灭绝组中的动物。重要的是，只有VNS改变了可塑性的IL和BLA之间的通路，当它被交付的消光环境。相比之下，迷走神经刺激传递到未受过训练的动物在其家笼对synapti无影响在所述IL-BLA途径（ 图4B）C可塑性。 <img alt="图1" src="/files/ftp_upload/53032/53032fig1.jpg" /> 图1.构建迷走神经袖口的。4毫米管件上的钻头的稳定的（A）的放置，显示了使用改进的针钻洞在油管。（B）的孔已在油管，和管已被砍倒的缝合线的中间。（℃）放置穿过箍的孔。注意，螺纹越过箍的嘴唇。铂铱丝到箍的（D）的插入。顶部显示完成接线，底部显示的布线。（E，F）与连接到导线末端的黄金引脚的完成袖口。见插入（F）的规模。（G）显示C在步骤3中描述的手术过程中被附着在头骨orresponding headcap件<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53032/53032fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/53032/53032fig2.jpg" /> 图2.迷走神经刺激和实验装置。（A）的设置了用于迷走神经刺激（VNS）示意图。动物经由headcap从中2铂iridum电线导致皮下到被缠绕在迷走神经的定做袖带电极连接到刺激隔离单元。周围的迷走神经袖带电极（B）的放置。用于设置的手术切口和袖带电极之前暴露的迷走神经的显微照片被缝合在其周围。（C）的照片听觉恐惧条件，具有连接到所述刺激器的动物。 （图来自参考20修改）。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53032/53032fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/53032/53032fig3.jpg" /> 图3. VNS增强听觉恐惧条件灭绝。（A）实验时间表。天1和2:听觉恐惧条件，8音（CS）/每天休克配对（美国）。第3天:在4未成基调演讲条件恐惧测试前，冻结测量。第4天:治疗，3组搭配VNS 4基调演讲，语气4演示搭配假刺激，加上假刺激（EE）20基调演讲。第5天:条件恐惧测试后，在4未成基调演讲冷冻测量（B）。 </strong&gt;的时间中所占比例介绍的条件刺激（CS），在第3天（D3，白色柱）和第5天（D5）用于不同治疗组中防冻（4种色调+假刺激，4种色调+ VNS，和EE ）。 1天灭绝培训搭配VNS后（黑色条）大鼠显著花费更少的时间比冰冻深水控制。邮政测试EE组（灰色条）冻结期间是类似4 +音调VNS组冻结。（C）的时间中所占比例在闭会期间音间隔（ITI）在第3天（D3，白条冻结）和第5天（D5）为B. VNS动物显示同样的群体也表现出降低空调的基调演讲之外冻结行为。 （* P &lt;0.05，误差棒代表平均值的标准误差）。 （图来自参考20修改）。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53032/53032fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 图4. VNS改变metaplasticity在所述IL-BLA途径。（A）的代表性的刺激，并在IL和BLA和记录在IL的血乳酸刺激后场电位。的输入-输出曲线的代表轨迹的记录位点（乙）突触可塑性的IL-BLA途径响应短脉冲刺激4组大鼠。右上:在恐惧空调大鼠IL短暂爆发刺激诱导有限公司在BLA。左中:接收4基调消退训练与假刺激老鼠也显示LTD。左下:鼠并获得扩展消退训练与假刺激显示没有变化，还是的LTD被诱导恐惧空调只组和假刺激组恢复。中间偏右:在VNS治疗的大鼠在单个分机inction会议，突触强度可进一步增强，导致LTP。右下图:VNS在家里笼交付导致的可塑性没有变化。简称:IL，infralimbic皮质; PL，前度皮质; BLA，杏仁核，洛杉矶，杏仁外侧核的基底外侧核; CE，杏仁中央核。 （* P &lt;0.05，误差棒代表平均值的标准误差）。 （图来自参考20修改）。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53032/53032fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图5" src="/files/ftp_upload/53032/53032fig5.jpg" /> 图5.简化脑干神经支配的迷走神经和二阶或更高阶的预测的示意图。通过其间接单胺核的调制在脑干，包括蓝斑（LC）和中缝nucle我（DRN），迷走神经可调节灭绝电路的重要组成部分，其中包括前额叶皮层（PFC），杏仁核（Amyg），海马（希普）和伏隔核（NAC）。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53032/53032fig5large.jpg" target="_blank">请点击这里查看一个更大的版本这个数字。</a>

Watch this Scientific Journal Video about Vagus Nerve Stimulation as a Tool to Induce Plasticity in Pathways Relevant for Extinction Learning at JoVE.com

Vagus Nerve Stimulation as a Tool to Induce Plasticity in Pathways Relevant for Extinction Learning

迷走神经刺激（VNS）已成为一种工具来促使针对性的突触可塑性的脑修改范围的行为。本协议描述了如何实现VNS方便灭绝的恐惧记忆的巩固。

迷走神经刺激作为一种工具来诱导可塑性相关途径灭绝学习

Extinction describes the process of attenuating behavioral responses to neutral stimuli when they no longer provide the reinforcement that has been ...

vagus-nerve-stimulation-as-tool-to-induce-plasticity-pathways

Nanyang Technological University

Research

JoVE Journal

Behavior

23.2K 次观看.  The University of Texas at Dallas. 迷走神经刺激（VNS）已成为一种工具来促使针对性的突触可塑性的脑修改范围的行为。本协议描述了如何实现VNS方便灭绝的恐惧记忆的巩固。 

视频: Vagus Nerve Stimulation as a Tool to Induce Plasticity in Pathways Relevant for Extinction Learning

Long-term Potentiation of Perforant Pathway-dentate Gyrus Synapse in Freely Behaving Mice

Studies of long-term potentiation of synaptic efficacy, an activity-dependent synaptic phenomenon having properties that make it attractive as a potential cellular mechanism underlying learning and information storage, have long been used to elucidate the physiology of various neuronal circuits in the hippocampus, amygdala, and other limbic and cortical structures. With this in mind, transgenic mouse models of neurological diseases represent useful platforms to conduct long-term&#160;potentiation (LTP) studies to develop a greater understanding of the role of genes in normal and abnormal synaptic communication in neuronal networks involved in learning, emotion and information processing. This article describes methodologies for reliably inducing LTP in the freely behaving mouse. These methodologies can be used in studies of transgenic and knockout freely&#160;behaving mouse models of neurodegenerative diseases.

Transgenic and knockout mouse models of neurological diseases are useful for studying the role of genes in normal and abnormal neurophysiology. This article describes methodologies which can be used to study long-term potentiation, a cellular mechanism which may underlie learning and memory, in transgenic and knockout freely&#160;behaving mouse models of neuropathology.

穿通通路 - 齿状回突触在自由活动的小鼠长时程增强

Studies of long-term potentiation of synaptic efficacy, an activity-dependent synaptic phenomenon having properties that make it attractive as a potential ...

科研

行为学

A Method for Remotely Silencing Neural Activity in Rodents During Discrete Phases of Learning

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while animals are engaged in learning and memory tasks. The approach combines pharmacogenetics (Designer-Receptors-Exclusively-Activated-by-Designer-Drugs) with a behavioral paradigm (sensory preconditioning) that is designed to distinguish between different forms of learning. Specifically, viral-mediated delivery is used to express a genetically modified inhibitory G-protein coupled receptor (the Designer Receptor) into a discrete brain region in the rodent. Three weeks later, when designer receptor expression levels are high, a pharmacological agent (the Designer Drug) is administered systemically 30 min prior to a specific behavioral session. The drug has affinity for the designer receptor and thus results in inhibition of neurons that express the designer receptor, but is otherwise biologically inert. The brain region remains silenced for 2-5 hr (depending on the dose and route of administration). Upon completion of the behavioral paradigm, brain tissue is assessed for correct placement and receptor expression. This approach is particularly useful for determining the contribution of individual brain regions to specific components of behavior and can be used across any number of behavioral paradigms.

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while rats are engaged in learning and memory tasks. The approach combines pharmacogenetics (Designer-Receptors-Exclusively-Activated-by-Designer-Drugs) with a behavioral paradigm (sensory preconditioning) that is designed to distinguish between different components of learning.

为远程沉默的神经活动在鼠害的方法在学习独立的阶段

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while animals are engaged in learning and ...

Extinction Training During the Reconsolidation Window Prevents Recovery of Fear

Fear is maladaptive when it persists long after circumstances have become safe. It is therefore crucial to develop an approach that persistently prevents the return of fear. Pavlovian fear-conditioning paradigms are commonly employed to create a controlled, novel fear association in the laboratory. After pairing an innocuous stimulus (conditioned stimulus, CS) with an aversive outcome (unconditioned stimulus, US) we can elicit a fear response (conditioned response, or CR) by presenting just the stimulus alone1,2 . Once fear is acquired, it can be diminished using extinction training, whereby the conditioned stimulus is repeatedly presented without the aversive outcome until fear is no longer expressed3. This inhibitory learning creates a new, safe representation for the CS, which competes for expression with the original fear memory4. Although extinction is effective at inhibiting fear, it is not permanent. Fear can spontaneously recover with the passage of time. Exposure to stress or returning to the context of initial learning can also cause fear to resurface3,4.
Our protocol addresses the transient nature of extinction by targeting the reconsolidation window to modify emotional memory in a more permanent manner. Ample evidence suggests that reactivating a consolidated memory returns it to a labile state, during which the memory is again susceptible to interference5-9. This window of opportunity appears to open shortly after reactivation and close approximately 6hrs later5,11,16, although this may vary depending on the strength and age of the memory15. By allowing new information to incorporate into the original memory trace, this memory may be updated as it reconsolidates10,11. Studies involving non-human animals have successfully blocked the expression of fear memory by introducing pharmacological manipulations within the reconsolidation window, however, most agents used are either toxic to humans or show equivocal effects when used in human studies12-14. Our protocol addresses these challenges by offering an effective, yet non-invasive, behavioral manipulation that is safe for humans.
By prompting fear memory retrieval prior to extinction, we essentially trigger the reconsolidation process, allowing new safety information (i.e., extinction) to be incorporated while the fear memory is still susceptible to interference. A recent study employing this behavioral manipulation in rats has successfully blocked fear memory using these temporal parameters11. Additional studies in humans have demonstrated that introducing new information after the retrieval of previously consolidated motor16, episodic17, or declarative18 memories leads to interference with the original memory trace14. We outline below a novel protocol used to block fear recovery in humans.

Conditioned fear can be diminished through an inhibitory process called extinction, but can resurface under conditions such as the passage of time or exposure to stress. Our protocol presents a novel way of preventing fear recovery by introducing extinction during the reconsolidation window (the re-storage phase of a reactivated memory).

灭绝培训在彭兰霞窗口无法还原的恐惧

Fear is maladaptive when it persists long after circumstances have become safe. It is therefore crucial to develop an approach that persistently prevents ...

神经科学

Ex Vivo Optogenetic Dissection of Fear Circuits in Brain Slices

Optogenetic approaches are now widely used to study the function of neural populations and circuits by combining targeted expression of light-activated proteins and subsequent manipulation of neural activity by light. Channelrhodopsins (ChRs) are light-gated cation-channels and when fused to a fluorescent protein their expression allows for visualization and concurrent activation of specific cell types and their axonal projections in defined areas of the brain. Via stereotactic injection of viral vectors, ChR fusion proteins can be constitutively or conditionally expressed in specific cells of a defined brain region, and their axonal projections can subsequently be studied anatomically and functionally via ex vivo optogenetic activation in brain slices. This is of particular importance when aiming to understand synaptic properties of connections that could not be addressed with conventional electrical stimulation approaches, or in identifying novel afferent and efferent connectivity that was previously poorly understood. Here, a few examples illustrate how this technique can be applied to investigate these questions to elucidating fear-related circuits in the amygdala. The amygdala is a key region for acquisition and expression of fear, and storage of fear and emotional memories. Many lines of evidence suggest that the medial prefrontal cortex (mPFC) participates in different aspects of fear acquisition and extinction, but its precise connectivity with the amygdala is just starting to be understood. First, it is shown how ex vivo optogenetic activation can be used to study aspects of synaptic communication between mPFC afferents and target cells in the basolateral amygdala (BLA). Furthermore, it is illustrated how this ex vivo optogenetic approach can be applied to assess novel connectivity patterns using a group of GABAergic neurons in the amygdala, the paracapsular intercalated cell cluster (mpITC), as an example.

Ex Vivo Optogenetic Dissection of Fear Circuits in Brain Slices

Optogenetic approaches are widely used to manipulate neural activity and assess the consequences for brain function. Here, a technique is outlined that&#160;upon in vivo expression of the optical activator Channelrhodopsin, allows for ex vivo analysis of synaptic properties of specific long range and local neural connections in fear-related circuits.

在脑片恐惧电路体外光遗传学解剖

Optogenetic approaches are now widely used to study the function of neural populations and circuits by combining targeted expression of light-activated ...

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 ...

Vagus Nerve Stimulation As an Adjunctive Neurostimulation Tool in Treatment-resistant Depression

Vagus nerve stimulation (VNS) is an approved neurostimulation therapy. The purpose of the method is to treat patients with therapy-resistant depression (TRD). VNS exhibits antidepressive and stabilizing effects. This method is particularly useful as a long-term treatment, in which up to two-thirds of patients respond. The vagus nerve stimulator is positioned on the left vagus nerve during a surgical procedure and is activated telemetrically by a wand connected to a handheld computerized device. The treating physician can perform various adjustments of the vagus nerve stimulator during in-office visits (e.g., by modifying stimulation intensity or stimulation frequency) to achieve maximum therapeutic effects with low side effects. Set-up of the device usually takes several months. Typical side effects include wound infection, temporary salivation, coughing, paralysis of the vocal cords, bradycardia, or even asystole. The patient can stop the VNS by placing a magnet over the generator. The current protocol describes delivery of the specific stimulation tool and methods for adjusting the tuning parameters to achieve the best remission rates in patients with TRD.

Vagus nerve stimulation (VNS) has been shown to be effective as an adjunct treatment for treatment-resistant depression (TRD). VNS leads to antidepressive and antisuicidal effects and quality of life improvements. This protocol offers a step-by-step guide to managing and adjusting a vagus nerve stimulator for efficient treatment of TRD.

迷走神经刺激作为抗抑郁抑郁症的辅助神经刺激工具

Vagus nerve stimulation (VNS) is an approved neurostimulation therapy. The purpose of the method is to treat patients with therapy-resistant depression ...

医学

Preparation of Peripheral Nerve Stimulation Electrodes for Chronic Implantation in Rats

Peripheral nerve cuff electrodes have long been used in the neurosciences and related fields for stimulation of, for example, vagus or sciatic nerves. Several recent studies have demonstrated the effectiveness of chronic VNS in enhancing central nervous system plasticity to improve motor rehabilitation, extinction learning, and sensory discrimination. Construction of chronically implantable devices for use in such studies is challenging due to rats&#8217; small size, and typical protocols require extensive training of personnel and time-consuming microfabrication methods. Alternatively, commercially available implantable cuff electrodes can be purchased at a significantly higher cost. In this protocol, we present a simple, low-cost method for construction of small, chronically implantable peripheral nerve cuff electrodes for use in rats. We validate the short and long-term reliability of our cuff electrodes by demonstrating that VNS in ketamine/xylazine anesthetized rats produces decreases in breathing rate consistent with activation of the Hering-Breuer reflex, both at the time of implantation and up to 10 weeks after device implantation. We further demonstrate the suitability of the cuff electrodes for use in chronic stimulation studies by pairing VNS with skilled lever press performance to induce motor cortical map plasticity.

Existing approaches for constructing chronically implantable peripheral nerve cuff electrodes for use in small rodents often require specialized equipment and/or highly trained personnel. In this protocol we demonstrate a simple, low-cost approach for fabricating chronically implantable cuff electrodes, and demonstrate their effectiveness for vagus nerve stimulation (VNS) in rats.

大鼠慢性植入的外周神经刺激电极的制备

Peripheral nerve cuff electrodes have long been used in the neurosciences and related fields for stimulation of, for example, vagus or sciatic nerves. ...

Ex Vivo Optogenetic Interrogation of Long-Range Synaptic Transmission and Plasticity from Medial Prefrontal Cortex to Lateral Entorhinal Cortex

Studying the physiological properties of specific synapses in the brain, and how they undergo plastic changes, is a key challenge in modern neuroscience. Traditional in vitro electrophysiological techniques use electrical stimulation to evoke synaptic transmission. A major drawback of this method is its nonspecific nature; all axons in the region of the stimulating electrode will be activated, making it difficult to attribute an effect to a particular afferent connection. This issue can be overcome by replacing electrical stimulation with optogenetic-based stimulation. We describe a method for combining optogenetics with in vitro patch-clamp recordings. This is a powerful tool for the study of both basal synaptic transmission and synaptic plasticity of precise anatomically defined synaptic connections and is applicable to almost any pathway in the brain. Here, we describe the preparation and handling of a viral vector encoding channelrhodopsin protein for surgical injection into a pre-synaptic region of interest (medial prefrontal cortex) in the rodent brain and making of acute slices of downstream target regions (lateral entorhinal cortex). A detailed procedure for combining patch-clamp recordings with synaptic activation by light stimulation to study short- and long-term synaptic plasticity is also presented. We discuss examples of experiments that achieve pathway- and cell-specificity by combining optogenetics and Cre-dependent cell labeling. Finally, histological confirmation of the pre-synaptic region of interest is described along with biocytin labeling of the post-synaptic cell, to allow further identification of the precise location and cell type.

Ex Vivo Optogenetic Interrogation of Long-Range Synaptic Transmission and Plasticity from Medial Prefrontal Cortex to Lateral Entorhinal Cortex

Here we present a protocol describing viral transduction of discrete brain regions with optogenetic constructs to permit synapse-specific electrophysiological characterization in acute rodent brain slices.

离体 从内侧前额叶皮层到外嗅皮层的远距离突触传递和可塑性的光遗传学询问

Studying the physiological properties of specific synapses in the brain, and how they undergo plastic changes, is a key challenge in modern neuroscience. ...

Using Optogenetics to Reverse Neuroplasticity and Inhibit Cocaine Seeking in Rats

This protocol demonstrates the steps needed to use optogenetic tools to reverse cocaine-induced plasticity at thalamo-amygdala circuits to reduce subsequent cocaine seeking behaviors in the rat. In our research, we had found that when rats self-administer intravenous cocaine paired with an audiovisual cue, synapses formed at inputs from the medial geniculate nucleus of the thalamus (MGN) onto principal neurons of the lateral amygdala (LA) become stronger as the cue-cocaine association is learned. We hypothesized that reversal of the cocaine-induced plasticity at these synapses would reduce cue-motivated cocaine seeking behavior. In order to accomplish this type of neuromodulation in vivo, we wanted to induce synaptic long-term depression (LTD), which decreases the strength of MGN-LA synapses. To this end, we used optogenetics, which allows neuromodulation of brain circuits using light. The excitatory opsin oChiEF was expressed on presynaptic MGN terminals in the LA by infusing an AAV containing oChiEF into the MGN. Optical fibers were then implanted in the LA and 473 nm laser light was pulsed at a frequency of 1 Hz for 15 minutes to induce LTD and reverse cocaine induced plasticity. This manipulation produces a long-lasting reduction in the ability of cues associated with cocaine to induce drug seeking actions.

The methods described here outline a procedure used to optogenetically reverse cocaine-induced plasticity in a behaviorally-relevant circuit in rats. Sustained low-frequency optical stimulation of thalamo-amygdala synapses induces long-term depression (LTD). In vivo optogenetically-induced LTD in cocaine-experienced rats resulted in the subsequent attenuation of cue-motivated drug seeking.

使用光遗传学逆转大鼠的神经可塑性并抑制可卡因的寻找

This protocol demonstrates the steps needed to use optogenetic tools to reverse cocaine-induced plasticity at thalamo-amygdala circuits to reduce ...

腹部 VNS 治疗大鼠模型中的炎症

Implantation Surgery for Abdominal Vagus Nerve Stimulation and Recording Studies in Awake Rats

Abdominal vagus nerve stimulation (VNS) can be applied to the subdiaphragmatic branch of the vagus nerve of rats. Due to its anatomical location, it does not have any respiratory and cardiac off-target effects commonly associated with cervical VNS. The lack of respiratory and cardiac off-target effects means that the intensity of stimulation does not need to be lowered to reduce side effects commonly experienced during cervical VNS. Few recent studies demonstrate the anti-inflammatory effects of abdominal VNS in rat models of inflammatory bowel disease, rheumatoid arthritis, and glycemia reduction in a rat model of type 2 diabetes. Rat is a great model to explore the potential of this technology because of the well-established anatomy of the vagus nerve, the large size of the nerve that allows easy handling, and the availability of many disease models. Here, we describe the methods for cleaning and sterilizing the abdominal VNS electrode array and surgical protocol in rats. We also describe the technology required for confirmation of suprathreshold stimulation by recording evoked compound action potentials. Abdominal VNS has the potential to offer selective, effective treatment for a variety of conditions, including inflammatory diseases, and the application is expected to expand similarly to cervical VNS.

作者聚焦:探索腹部 VNS 治疗炎症性疾病

The present protocol describes the surgical technique for implanting an electrode array onto the abdominal vagus nerve in rats, along with methods for chronic electrophysiology testing and stimulation using the implanted device.

腹部迷走神经刺激植入手术和清醒大鼠的记录研究

Abdominal vagus nerve stimulation (VNS) can be applied to the subdiaphragmatic branch of the vagus nerve of rats. Due to its anatomical location, it does ...

迷走神经刺激作为一种工具来诱导可塑性相关途径灭绝学习

摘要

探索更多视频

此视频中的章节

穿通通路 - 齿状回突触在自由活动的小鼠长时程增强

为远程沉默的神经活动在鼠害的方法在学习独立的阶段

灭绝培训在彭兰霞窗口无法还原的恐惧

在脑片恐惧电路体外光遗传学解剖

成人脑切片长期抑郁症诱导评估

迷走神经刺激作为抗抑郁抑郁症的辅助神经刺激工具

大鼠慢性植入的外周神经刺激电极的制备

离体从内侧前额叶皮层到外嗅皮层的远距离突触传递和可塑性的光遗传学询问

使用光遗传学逆转大鼠的神经可塑性并抑制可卡因的寻找

腹部迷走神经刺激植入手术和清醒大鼠的记录研究

迷走神经刺激作为一种工具来诱导可塑性相关途径灭绝学习

摘要

探索更多视频

此视频中的章节

穿通通路 - 齿状回突触在自由活动的小鼠长时程增强

为远程沉默的神经活动在鼠害的方法在学习独立的阶段

灭绝培训在彭兰霞窗口无法还原的恐惧

在脑片恐惧电路体外光遗传学解剖

成人脑切片长期抑郁症诱导评估

迷走神经刺激作为抗抑郁抑郁症的辅助神经刺激工具

大鼠慢性植入的外周神经刺激电极的制备

离体 从内侧前额叶皮层到外嗅皮层的远距离突触传递和可塑性的光遗传学询问

使用光遗传学逆转大鼠的神经可塑性并抑制可卡因的寻找

腹部迷走神经刺激植入手术和清醒大鼠的记录研究

离体从内侧前额叶皮层到外嗅皮层的远距离突触传递和可塑性的光遗传学询问