Name: combined shuttle-box training with electrophysiological cortex recording and stimulation as a tool to study perception and learning
Uploaded: 2015-10-22T15:00:00.000+00:00
Duration: 8 min 43 s
Description: Watch this Scientific Journal Video about Combined Shuttle-Box Training with Electrophysiological Cortex Recording and Stimulation as a Tool to Study Perception and Learning at JoVE.com

这项工作是由来自Deustche研究联合会DFG和莱布尼茨研究所神经生物学的资助。我们感谢玛丽亚 - 滨海Zempeltzi和凯萨琳·奥尔的技术援助。

在这项工作中提出的所有实验均与由德国法律对实验动物的保护规定的道德标准的协议进行的。实验批准了萨克森 - 安哈尔特州的伦理委员会。 1.订造多通道电极阵列的微刺激和记录<ol><li>客户定制微刺激阵列<ol><li>对于提供ICMS，准备刺激电极使用3厘米长的聚四氟乙烯绝缘不锈钢丝所需的空间设计（2个通道位置横向阵列）（O带隔离= 50微米）。 见图2。 </li><li>压接电线的一端与阳针形系统（1.25毫米间距）。 </li><li>对于2信道数组〜0.5mm或0.7〜毫米电极距离通电线垂直地通过两个垂直排列的电子显微镜对象保持器网格（网格距离〜5毫米的）引导电线（0.1654毫米节距）。 </li><li&gt;胶导线一起用一小滴的牙科丙烯酸约4mm以上终于作为电极尖端的导线的结局。 </li><li>把男性销插入插头外壳。 </li><li>把电极与电解质溶液（例如，0.9％氯化钠）的盘中，测量每个信道的与各自的阻抗测量装置的阻抗（例如，FHC阻抗调节模块）。瞄准在100〜500千欧的阻抗范围。 <ol><li>如果阻抗太高，递送短的电流（1秒，以1mA）通过通道以降低它。再次检查阻抗。 </li></ol></li></ol></li><li>特制的高分辨率多通道录音阵列 注:本协议描述制造表面阵列记录脑电（ECOG）数据。然而，设计可定制，以满足的各研究问题的要求（深度录音等等）。高分辨率多通道阵列BUIL牛逼由聚四氟乙烯绝缘的不锈钢丝（直径带隔离= 50微米）压接至插针系统（1.25 mm间距）。 <ol><li>为表面电极阵列，引导导线18通过两个制备的电子显微镜对象保持器网格的3×6矩阵。 </li><li>嵌入用牙科丙烯酸所述栅格之间的导线的6×3排列，并把销在插头外壳中。 </li><li>通过使用粉碎机附着到手持电钻研磨牙科丙烯酸成矩形块。 </li><li>检查所有通道的阻抗（参见1.1.6）是在100的范围内 - 500千欧。 </li><li>植入前（见步骤2）阵列的研磨表面以匹配皮质表面的凸。 </li></ol></li></ol>阵列到听觉皮层在麻醉蒙古Herbils慢性用法2.手术植入<ol><li>握住正式批准执行计划的所有必要的实验步骤。穿Appro公司priate防护服（上衣，无菌手套，口罩，头罩）。 </li><li>使用成年雄性长爪沙鼠（ 长爪沙鼠 ）或任何其他啮齿动物。戴防护手套和外套，而应使用消毒，以及确定的外科手术工具。 </li><li>万一沙鼠，麻醉动物用腹腔注射氯胺酮（100毫克/千克）和赛拉嗪（5mg / kg的）稀释在0.9％无菌氯化钠溶液的混合物。甲苯噻嗪::氯化钠（0.9％）中的9:1的比例:10通过输注的0.6毫升/小时/千克氯胺酮的混合物组成的维护。 </li><li>放置动物在加热板上用于通过直肠探头反馈系统维持体温在37℃下（例如，世界精密仪器）。 </li><li>刮皮毛覆盖interoccipital，顶叶和颞骨。无菌制备切口部位用有效的消毒剂（例如，交替的优碘或Nolvasan和70％的酒精擦洗3次）。镨努力保护眼睛对抗干燥用眼膏。 </li><li>切开颅骨皮肤覆盖的interoccipital，顶叶和额叶用手术刀骨骼和轻轻移动钻头在骨头表面去除骨膜。 </li><li>钻在对侧顶叶和额叶骨的外侧尽可能小的孔，以不妨碍以后的头安装。现在螺杆两个骨螺钉的直径为约1mm紧密地插入孔中。确保该壁层螺杆具有良好的接触，以硬脑膜，因为这将在后面用作公共参考和接地，以及用于电刺激返回电极。 </li><li>内侧粘上小铝棒在额骨和植入由headh​​older过程中使用它作为头部固定。 </li><li>去除皮肤覆盖在家鼠颞一侧用剪刀。 </li><li>切断颞肌的背侧，以获得入场颞骨。 </li><li>通过开颅手术（〜3毫米x 4 mm以内）暴露听觉皮层通过使用牙钻的颞骨。识别的基础上典型血脑血管签名18,19听觉皮层的位置。 </li><li>小心使硬脑膜切开用微手术刀在那里的刺激电极被植入大脑的位置。轻轻沿表面移动直到硬脑膜泪禁止损坏底层神经纤维。 </li><li>将在显微控制刺激阵列。选择的切向插入角度，以允许硬膜外阵列被放置在感兴趣的区域。根据插入角度及位置和刺激的目标位，仔细考虑插入深度。 </li><li>放置具有良好的接触，以脑表面通过第二显微覆盖感兴趣的区域（这里听觉皮层）硬膜外表面记录阵列。注意使阵列皮层的曲率相匹配，以不缩进硬脑膜。 </li><li>固定两个电极阵列及其插头外壳用牙科丙烯酸的头骨。 </li><li>覆盖暴露的皮质表面防腐润滑剂（ 如 KY-果冻），靠近开颅牙科丙烯酸（ 例如，Paladur，贺利氏Ulzer）。请记住，聚合会产生热量。使用神经纤维和腈纶牙之间有足够的润滑，防止牙丙烯酸和皮质表面的任何接触，因为这可能会造成组织损伤。 </li><li>现在，让动物的任何进一步的程序开始前恢复。恢复时间可能种（:≥3天，老鼠:≥7天如，沙鼠 ）之间的差异。仔细监测动物的状态，并给予止痛治疗，在接下来的日子里，如果适用（ 例如，美洛昔康; 1毫克/千克手术后和手术后24小时）。 </li></ol> 3.双向穿梭箱设计中使用ICMS作为条件刺激<ol><li>穿梭箱训练<ol><li>将一梭箱（Custom建造或任何商业产品， 例如，E10，E15，Coulbourn仪器）在声学和电屏蔽室。盒子包含两个舱室的一道坎分开。考虑障碍的高度，因为它影响的复响应的行为偏差。使用的具体品种适当高度（ 例如，〜2厘米老鼠〜3个厘米长爪沙鼠）。 </li><li>要应用足部电击（美国），使用网格地板适用于在调查12,15物种酒吧之间的距离。务必要小心，没有电的快捷方式单独杆（粪便，毛发，电极膏）。 </li><li>动物（见2.17）有相当的恢复时间后，连接受制于记录和刺激电缆最好不使用任何短期的麻醉。试图掩盖用毛巾动物，轻轻地走动物手中。揭开用另一只手的动物的头部和连接器和连接电缆。 </li&gt; <li>让动物使其习惯了训练室每次会议开始前3分钟。 </li><li>每堂训练课（1 - 2日报）申请的30 - 90试验。高达15秒的反复试验持续时间，以及随机变化的25间审间的时间间隔 - 30秒。 </li><li>对于ICMS提供经调节的刺激，刺激电极使用多通道刺激（ 例如，MCS STG2000）。要获取行为效应不破坏脑组织，应用脉冲序列（ 例如，300毫秒长，100 PPS）的双相，电荷平衡脉冲（阴极第一）与200微秒阶段的持续时间。为700毫秒，持续4秒（观察窗）的持续时间的暂停重复列车。 </li><li>为了呈现听觉CS使用模拟输出的PCI板卡（如NI PCI-6733）。这些设备编写程序用Matlab进行灵活的控制，并通过数字输出线的硬件触发穿梭箱系统。 </li><li>路线模拟输出信号为th通过音频放大器e快速箱扬声器。 </li><li>有条件交付通过格地板足部电击美国。为了获得最佳的电生理记录的质量，产生的震动由第二高端多通道刺激（MCS STG2000）。 </li><li>要应用检测培训目前只有CS +的刺激。这里的测试试验（〜10％）之间散布未经CS和美国本空白试验以校正偏置梭行为（参见3.2）。为了更艰巨的任务，训练动物CS +，并在同一会话随机排序的CS-刺激区分。 </li><li>分类4秒钟内（CR）作为命中响应的临界时间窗的CS +发病后的隔室变化。在关键的时间窗口（ 小姐）无CR CS +试验，立即将温和足部冲击6〜10秒的条件刺激（美国）。 注:重复CS +的刺激，与美国的重叠会降低学习努力的动物ND提高学习速度和性能（&#39;延迟&#39;与&#39;跟踪&#39;调理，见讨论）。 </li><li>对于CS-试验，分类的关键时间窗口， 误报响应内的隔间的变化，而 ​​这种不恰当的CR后立即申请了美国长达10秒。 CS-后当动物在关键的时间窗口期间留在车厢（无CR）不适用美国，而这个审判列为正确的拒绝 。 注:重要的是，脚休克美国始终处于关机状态，当动物的应对之道（ 越狱 ）的变化舱。使用较长的关键时间窗CS-试验， 例如，如果重复的条件刺激时，尽管这会判处更保守的学习标准，因为它提供了更高的动物为了抑制CR。 </li><li>对于CS和美国的有效关联调整的冲击强度在中等范围，以便厌恶，但不痛。最佳初始l，电流强度物种之间的差异（例如，50μA的老鼠，200μA为长爪沙鼠）。因此，请参阅下两发子弹做进一步的技术细节: <ol><li>分别调整震荡的实力，在第一堂训练课开始用温和的幅度（约200μA为长爪沙鼠）。如果脚部的冲击力实在是太低了，潜伏期抖动和协会之间的CS和美国是不是最佳的。 </li><li>总是注意如果动物开始发声以及响应于所述CS到冻结。在这种情况下，电击强度太高。这种条件性恐惧反应干扰了逃避学习。 </li></ol></li><li>仔细确定潜伏​​期。增加足部冲击力一步一步，如果潜伏期长于2秒第20次试验后。检查动物是在冲击的控制， 即每训练后，就说明潜伏期大大低于2秒。 注:然而，避免增加脚震荡走强太快，作为一个高度强调动物的行为策略可以回落到纯粹的逃避反应。因此密切观察同时调整美国的振幅行为，尤其是反应潜伏期。举一个例子见图3E。 </li><li>如果动物显示CR，立即停止介绍CS的。这是为了回避反应的加强是至关重要的。 </li><li>如果动物已经获得了回避的策略， 即显示出相应的CR于所有CS，改变参数（ICMS幅度，相位时间，重复率等）来进行心理分析。在一列块的方式与美国配对，但是这将导致学习和适应应用参数CS变化。 <ol><li>为了避免这种情况，开始与原来的训练15至30次试验，然后随机点缀CS变化没有美国为与原CS的正规训练试验的测试试验。最大üSE 25测试试验％。 </li></ol></li><li>培训结束后从培养室取出的动物和下一个动物训练之前，请仔细清洗完成框。尽量避免在同一个盒子一个时间的训练不同的物种，它们的自然气味可能与培训绩效干扰。 </li></ol></li><li>训练数据的分析<ol><li>记录的习惯化阶段的所有车厢的变化。 </li><li>记录在培训期间的所有车厢的变化，分成命中和误报，逃跑反应（未命中和背部班车后，一场虚惊）和自发intertrial班车服务（ITS）。 </li><li>计算出CR率CS +和CS如下:命中率=命中/ CS的+试验次数;误报率=误报/的CS - 试验编号。 </li><li>获得CR率会明智的。然而，以评估学习动态以较高的时间分辨率，从CS +和CS试验相同数量的短块计算CR率（ <e米&gt;如，10例），分别。 </li><li>剧情CR率作为会话或试块，用于评估训练进度和学习动力的功能。 </li><li>对于行为灵敏度独立的实验条件偏压动物的反应进行定量，导出基于信号检测理论8,9,17 D&#39;值。 </li><li>对于D&#39;分析利用Z分数相应命中和误报率（辨别学习），或打从一个标准正态分布函数的逆派生率和它（检测学习），并减去这些z值。设置一个阈值标准为刺激检测的D&#39;= 1.0，其对应于上述噪声的标准偏差的信号的强度。 参见图3作为例子。 </li><li>另外确定CR-并通过测量CS发作和行为反应之间的时间周期逸出的反应时间响应于不同的CS（完全隔室改变）。 </li><li>从偷拍穿梭箱行为获取更详细的行为分析。从视频中的音频迹穿梭箱或刺激系统记录触发脉冲实现视频和录像系统之间的时间同步。视频分析允许之前的CR动物驴的关注和定向反应。 </li></ol></li></ol> 4.在学习动物体内电生理技术<ol><li>在训练过程中电生理记录<ol><li>在训练期间，记录电生理信号（例如，所描述的脑电图阵列）从多个电极monopolarly反对共同参考/接地电极。 </li><li>从所有的电极变成探头放大器馈送信号直接或经由短适配器插入头连接器堵塞。 </li><li>通过薄，软电缆线束包裹连接探头到主放大器由金属网咬动物保护它免受损坏。 注:在电缆线束机械压力可以通过弹簧进一步允许在框中动物的自由移动和旋转得到缓解。理想是一个可转动和电动旋转的使用。然而，对于听觉实验将旋转的隔音室外面或音盾用泡沫来减少它的电机产生的高频听觉噪声。 </li><li>使用在屏蔽盒的前置放大器，以增加信噪比和带通滤波器的信号中的期望频率范围。 </li><li>超过1千赫取样频率（用于局部场电位记录）和至少40千赫（用于动作电位记录）样本数据并存储到PC用于离线分析。使用适当的过滤器设置两种类型 （如2 - 300赫兹的局部领域的潜力; 300 - 4000赫兹的动作电位）。 </li><li> T的开始前仔细检查记录的质量下雨（没有噪音或运动伪影）。应用在线FFT的滤波器对信号来确定的50 Hz的噪声的振幅。如果有必要仔细检查头连接件之间的连接，适配器的的探头，电缆线束，和放大器。 </li><li>对于在由电刺激诱发的记录数据减少伪像使用一个内插程序，以重建直到〜5毫秒各脉冲的发病后1毫秒前受工件的所有数据点。对于这一点，插入零的未受影响的数据点之间，并应用一种对称FIR滤波器最小化的内插点和它们的理想值（Matlab的interp.m功能）之间的均方误差。应用此方法对原始信号，分别为前进一步分析9每个通道。 </li></ol></li><li>并行穿梭盒子训练，ICMS，和录音技术细节<ol><li>一般情况下，保证动物感到舒适框surroundiNG。让动物自由移动，到达箱的各个角落。足够的时间第一次训练（20分钟），并且每个会议前（3分钟）才使其习惯一天是有益的。 </li><li>经过任何手术治疗，以便在必要时（见上文2.17）的动物有足够的时间来恢复，包括药物治疗，并开始只是训练的动物，如果动物没有显示遭受或痛苦的任何典型症状（紧闭双眼，昏昏欲睡的表型，褴褛的皮毛20）。 </li><li>保证电网地板的正确接地。必须避免记录系统，穿梭箱和动物之间的接地环路。只能通过其共同的接地电极接地动物，离开gridfloor在浮充电压。 </li><li>多通道刺激（MCS STG2000）连接到通过电动旋转的单独的行植入刺激电极阵列的接头。 </li><li>使用记录的共同接地电极作为接地或返回电极的余CMS，也是如此。 </li></ol></li></ol>电极位置5.组织学分析<ol><li>以下完整训练集，控制用于通过组织学分析的刺激电极阵列的稳定位置。 </li><li>麻醉动物用腹腔注射氯胺酮（100毫克/千克）和赛拉嗪（5mg / kg的）稀释在无菌钠氯化物0.9％的混合物。然后应用单极阴极电流（30μA，持续60秒）通过各种刺激渠道提供获得铁矿床，在组织上植入8的位置 。 </li><li>根据这一程序，通过安乐死适当和认可的方法（100毫克/公斤，例如腹腔注射戊巴比妥过量的）牺牲的动物。 </li><li>立即取出该动物的大脑和冻结它在2-甲基丁烷冷却至-70℃，在液氮中。 </li><li>现在削减对低温恒温器切片机感兴趣的区域为50微米^ horizo​​ntal部分。 </li><li>组织学:尼氏和"普鲁士蓝"-staining <ol><li>为了识别皮质层的对待每一个第二片与尼氏染色。首先，洗澡切片5分钟，0.05M乙酸钠三水合物缓冲液（pH4.0 -4.2）。 </li><li>洗澡片5 - 在5％甲酚紫醋酸盐10分钟。用蒸馏水冲洗切片。 </li><li>沐浴在0.05M乙酸钠三水合物缓冲液（pH 4.0 - 4.2）2分钟连续切片，并在50％，70％和90％的乙醇，分别解决方案。 </li><li>在异丙醇洗澡片两次:96％乙醇（2:1）各5分钟。 </li><li>最后，洗澡切片Roticlear 5分钟三次。 </li><li> ，每隔片，使铁沉积可见实验后诱发的长单相电流的"普鲁士蓝"-staining获取的刺激通道位置。 </li><li>制备的1％的钾hexacyanoferrat新鲜溶液（Ⅱ）三水合物K 4[铁（CN）6]通过混合2克K 4铁[Fe（CN）6]在200ml 1％HCl中。 </li><li>加入800毫升0.1M磷酸缓冲液（pH7.4）中。 </li><li>洗澡脑切片10分钟，用蒸馏水，然后在hexacyanoferrat溶液10分钟。 </li><li>洗澡切片两次在0.1M磷酸盐缓冲液10分钟，最后5分钟在蒸馏水中。 </li></ol></li></ol>

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Electrophysiological mapping of frequency representation and distinction of fields.</a> The European journal of neuroscience. 5, 882-897 (1993).</li><li>Budinger, E., Heil, P., Scheich, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+organization+of+auditory+cortex+in+the+Mongolian+gerbil+(Meriones+unguiculatus).+III.+Anatomical+subdivisions+and+corticocortical+connections.">Functional organization of auditory cortex in the Mongolian gerbil (Meriones unguiculatus). III. Anatomical subdivisions and corticocortical connections.</a> European Journal of Neuroscience. 12, 2425-2451 (2000).</li><li>Langford, D. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coding+of+facial+expressions+of+pain+in+the+laboratory+mouse.">Coding of facial expressions of pain in the laboratory mouse.</a> Nature Methods. 7 (6), 447-449 (2010).</li><li>Ohl, F. W., Scheich, H., Freeman, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Change+in+pattern+of+ongoing+cortical+activity+with+auditory+category+learning.">Change in pattern of ongoing cortical activity with auditory category learning.</a> Nature. 412 (6848), 733-736 (2001).</li><li>Scheich, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+semantics+of+learning+and+crossmodal+processing+in+auditory+cortex:+the+semantic+processor+concept.">Behavioral semantics of learning and crossmodal processing in auditory cortex: the semantic processor concept.</a> Hearing Research. 271 (1-2), 3-15 (2011).</li><li>Happel, M. F. K., Jeschke, M., Ohl, F. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spectral+integration+in+primary+auditory+cortex+attributable+to+temporally+precise+convergence+of+thalamocortical+and+intracortical+input.">Spectral integration in primary auditory cortex attributable to temporally precise convergence of thalamocortical and intracortical input.</a> The Journal of Neuroscience. 30 (33), 11114-11127 (2010).</li><li>Ranck, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Which+elements+are+excited+in+electrical+stimulation+of+mammalian+central+nervous+system:+a+review.">Which elements are excited in electrical stimulation of mammalian central nervous system: a review.</a> Brain Research. 98, 417-440 (1975).</li><li>Clark, K. L., Armstrong, K. M., Moore, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Probing+neural+circuitry+and+function+with+electrical+microstimulation.">Probing neural circuitry and function with electrical microstimulation.</a> Proceedings. Biological sciences / The Royal Society. 278 (1709), 1121-1130 (2011).</li><li>Ilango, A., Shumake, J., Wetzel, W., Scheich, H., Ohl, F. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+stimulation+of+lateral+habenula+during+learning:+frequency-dependent+effects+on+acquisition+but+not+retrieval+of+a+two-way+active+avoidance+response.">Electrical stimulation of lateral habenula during learning: frequency-dependent effects on acquisition but not retrieval of a two-way active avoidance response.</a> PloS one. 8 (6), e65684 (2013).</li><li>Weible, A. P., McEchron, M. D., Disterhoft, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cortical+involvement+in+acquisition+and+extinction+of+trace+eyeblink+conditioning.">Cortical involvement in acquisition and extinction of trace eyeblink conditioning.</a> Behavioral Neuroscience. 114, 1058-1067 (2000).</li><li>Rothe, T., Deliano, M., Scheich, H., Stark, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Segregation+of+task-relevant+conditioned+stimuli+from+background+stimuli+by+associative+learning.">Segregation of task-relevant conditioned stimuli from background stimuli by associative learning.</a> Brain Research. 1297, 143-159 (2009).</li></ol>

作者什么都没有透露。

此协议通过使用双向厌恶足部电击控制穿梭盒子系统描述同时位点特异性ICMS和多通道电生理记录在一个学习的动物的方法。协议强调技术的关键概念为这样的组合，并指出只有通过它的共同接地电极接地的动物，留下gridfloor在浮动电压的重要性。在这里，听梭箱学习应用于长爪沙鼠的学习相关的塑料重组这些动物的听觉皮层中得到了广泛的8,12,14,15,21,22研究。然而，所描述的协议可以适于...

行为神经科学的一个基本目标是建立神经元的结构和功能特性，学习和认知之间的具体联系。与感知和学习有关的神经活性可以通过在多个位点的动作电位并局部场电位的各种大脑结构电生理记录进行研究。尽管电生理记录提供的神经活动和行为之间的相关协会，直接电皮层内微刺激（ICMS）一个多世纪以来一直是最直接的方法神经元的兴奋的人群和他们的行为和感知效果1的测试因果关系- 3。许多研究表明，动物是能够利用电刺激的知觉的任务范围内，例如初级视4吨取决于刺激位置的各种空间和时间属性onotopic 5，或躯体6区皮层。在皮层电诱发活动的传播主要是通过轴索纤维及其分布的突触连接2，在皮质，显然是一层依赖7的布局。得到的多突触活化ICMS诱发是今后更广泛的比电场2,8,9的直接影响。这就解释了为什么感性效应阈值由皮质内微刺激诱 ​​发可以强烈地层依赖8,10,11和站点相关的9。最近的一项研究证明了细节，上层的刺激产生的主要supragranular层corticocortical电路更广泛的激活，而皮质导致更深层的一个焦点，反复corticoefferent intracolumnar激活刺激。并行行为实验表明，后者具有低得多的听觉检测苏氨酸esholds 8。因此，位点特异性ICMS作为条件刺激的优点是利用与电生理记录结合因果涉及特定皮质电路激活8学习和感知在穿梭箱行为的措施。 双向穿梭箱范式是研究回避学习12一个完善的实验室设备。梭箱包括2个舱室的一道坎或门口分开。甲条件刺激（CS），其通过适当的信号象光或声音来表示，是偶然随后厌恶无条件刺激（US），如例如一个足部电击在金属网格地板。受试者可以学习，以避免与美国通过从一个穿梭箱隔室中响应于所述CS穿梭到其他。穿梭箱学习涉及区分学习阶段的序列13,14:首先，科学预测，美国从CS由经典条件和从美国逃出用仪器调节，因为美国是在穿梭终止。在下一阶段，科目学习完全避免美国通过穿梭于应对美国发病前的CS（ 回避反应）。一般情况下，穿梭箱学习包括经典性条件反射，有助于调理，以及根据学习阶段14的目标导向的行为。 梭箱程序，可以方便地建立，一般经过几次的日常训练课15产生强大的行为- 17。除了简单的逃避性（检测），穿梭箱可进一步用于通过采用GO / NOGO范式研究刺激歧视。这里，训练动物，以避免与美国由一个条件反应（CR）（去行为;梭成相对隔室）响应于&lt;STRONG&gt;去刺激（CS +）和NOGO行为 （留在当前车厢;无CR）响应NOGO刺激（CS-）并行微刺激和记录神经活动的高密度多电极阵列允许学习。的生理机制成功的学习标的。几个技术细节，根本的穿梭盒子训练，ICMS和平行电成功的组合，将被讨论。

<table><tbody><tr><td>Teflon-insulated stainless steel wire</td><td>California Fine Wire</td><td></td><td>diam. 50&amp;micro;m w/ isolation</td></tr><tr><td>Pin connector system&amp;nbsp;</td><td>Molex Holding GmbH</td><td>510470200</td><td>1.25 mm pitch PicoBlade</td></tr><tr><td>TEM grid Quantifoil</td><td>Science Services</td><td>EQ225-N27</td><td></td></tr><tr><td>Dental acrylic Paladur</td><td>Heraeus Kulzer</td><td>64707938</td><td></td></tr><tr><td>Hand-held drill OmniDrill35</td><td>WPI&amp;nbsp;</td><td>503599</td><td></td></tr><tr><td>Ketamine 500mg/10ml</td><td>Ratiopharm GmbH</td><td>7538837</td><td></td></tr><tr><td>Rompun 2%, 25ml</td><td>Bayer Vital GmbH</td><td>5066.0</td><td></td></tr><tr><td>Sodium-Chloride 0.9%, 10ml</td><td>B.Braun AG&amp;nbsp;</td><td>PRID00000772</td><td></td></tr><tr><td>Lubricant KY-Jelly</td><td>Johnson &amp;amp; Johnson</td><td></td><td></td></tr><tr><td>Shuttle-box E10-E15</td><td>Coulbourn Instruments</td><td>H10-11M-SC</td><td></td></tr><tr><td>Stimulus generator MCS STG 2000</td><td>Multichannel Systems</td><td></td><td></td></tr><tr><td>Plexon Headstage cable 32V-G20</td><td>Plexon Inc.</td><td>HSC/32v-G20</td><td></td></tr><tr><td>Plexon Headstage&amp;nbsp; 32V-G20</td><td>Plexon Inc.</td><td>HST/32v-G20</td><td></td></tr><tr><td>PBX preamplifier 32 channels</td><td>Plexon Inc.</td><td>32PBX box</td><td></td></tr><tr><td>Multichannel Acquisition System</td><td>Plexon Inc.</td><td>MAP 32/HLK2</td><td></td></tr><tr><td>Cryostate CM3050 S</td><td>Leica Microsystems GmbH</td><td></td><td></td></tr><tr><td>Signal processing Card Ni-Daq</td><td>National Instruments</td><td></td><td></td></tr><tr><td>Lab StandardTM Stereotaxic Instruments</td><td>Stoelting Co.&amp;nbsp;</td><td></td><td></td></tr><tr><td>Audio attenator g.pah</td><td>g.pah Guger technologies</td><td></td><td></td></tr><tr><td>Cresyl violet acetate</td><td>Roth GmbH</td><td>7651.2</td><td></td></tr><tr><td>Roticlear&amp;nbsp;</td><td>Roth GmbH</td><td>A538.1</td><td></td></tr><tr><td>Sodium acetate trihydrate</td><td>Roth GmbH</td><td>6779.1</td><td></td></tr><tr><td>Potassium hexacyanoferrat(II) trihydrate</td><td>Roth GmbH</td><td>7974.2</td><td></td></tr><tr><td>Di-sodium hydrogen phospahte dihydrate</td><td>Merck</td><td>1,065,801,000</td><td></td></tr><tr><td>ICM Impedance Conditioning Module</td><td>FHC</td><td>55-70-0</td><td></td></tr><tr><td>Animal Temperarture Controler</td><td>World Precision Instruments</td><td>ATC2000</td><td></td></tr></tbody></table>

combined shuttle-box training with electrophysiological cortex recording and stimulation as a tool to study perception and learning

穿梭箱回避学习是行为神经科学和实验设置一个行之有效的方法是传统的定制;必要的设备现在可以通过一些商业公司。该协议提供了一个双向穿梭箱回避学习范式的啮齿类动物的详细说明（在这里长爪沙鼠， 长爪沙鼠 ）的结合位点特异性电气皮层内微刺激（ICMS），并同时慢性电在体内录音。详细的协议，适用于研究在不同的啮齿类动物的学习行为和感知的多个方面。 听觉皮层电路如下条件刺激的位点特异性ICMS作为工具来测试特定的传入，传出和皮质内连接的感知相关。不同的活动模式可以通过使用不同的刺激电极ARR被诱发AYS的地方，层依赖ICMS或远处ICMS网站。利用行为信号检测分析可以判断哪个刺激策略是最有效的用于引发一个行为上可检测的和显着信号。此外，使用不同的电极设计（表面电极，深部电极等）并行多信道-录音允许对这种学习过程的时间过程研究神经元观测。这将是讨论的行为设计的变化是如何能提高认知的复杂性（ 如探测，识别，逆转学习）。

本节介绍穿梭盒子学习在蒙古沙鼠一个代表性的例子。受试者被训练来区分2激励电极之间的ICMS站点植入700微米彼此分离在听觉皮层（图1和2）。刺激阵列可定制在不同的空间设计（ 图1）。在这里，两个ICMS站点歧视与内呈现30 CS +和CS每次（ 图3A-C）次训练的经验教训。这是通过的命中和误报反应的完全缓解率在整个连续7个培训班（ 图3B）</...

Watch this Scientific Journal Video about Combined Shuttle-Box Training with Electrophysiological Cortex Recording and Stimulation as a Tool to Study Perception and Learning at JoVE.com

Combined Shuttle-Box Training with Electrophysiological Cortex Recording and Stimulation as a Tool to Study Perception and Learning

穿梭箱回避学习是行之有效的行为神经。本协议描述了如何穿梭盒子学习啮齿类动物可以与特定地点的电气皮层内微刺激（ICMS） 和体内录音作为研究学习和感知的多个方面的工具的同时慢性相结合。

联合穿梭盒子训练与电的Cortex记录和刺激作为一种工具来研究感知和学习

Shuttle-box avoidance learning is a well-established method in behavioral neuroscience and experimental setups were traditionally custom-made; the ...

combined-shuttle-box-training-with-electrophysiological-cortex

Universidad San Sebastian

Research

JoVE Journal

Behavior

10.2K 次观看.  Leibniz Institute for Neurobiology, Magdeburg, Germany. 穿梭箱回避学习是行之有效的行为神经。本协议描述了如何穿梭盒子学习啮齿类动物可以与特定地点的电气皮层内微刺激（ICMS） 和体内录音作为研究学习和感知的多个方面的工具的同时慢性相结合。 

视频: Combined Shuttle-Box Training with Electrophysiological Cortex Recording and Stimulation as a Tool to Study Perception and Learning

An Experimental Platform to Study the Closed-loop Performance of Brain-machine Interfaces

The non-stationary nature and variability of neuronal signals is a fundamental problem in brain-machine interfacing. We developed a brain-machine interface to assess the robustness of different control-laws applied to a closed-loop image stabilization task. Taking advantage of the well-characterized fly visuomotor pathway we record the electrical activity from an identified, motion-sensitive neuron, H1, to control the yaw rotation of a two-wheeled robot. The robot is equipped with 2 high-speed video cameras providing visual motion input to a fly placed in front of 2 CRT computer monitors. The activity of the H1 neuron indicates the direction and relative speed of the robot's rotation. The neural activity is filtered and fed back into the steering system of the robot by means of proportional and proportional/adaptive control. Our goal is to test and optimize the performance of various control laws under closed-loop conditions for a broader application also in other brain machine interfaces.

We use a closed-loop fly-machine interface to investigate general principles in neuronal control.

研究脑机接口的闭环性能的实验平台

The non-stationary nature and variability of neuronal signals is a fundamental problem in brain-machine interfacing. We developed a brain-machine ...

科研

神经科学

Modulating Cognition Using Transcranial Direct Current Stimulation of the Cerebellum

Numerous studies have emerged recently that demonstrate the possibility of modulating, and in some cases enhancing, cognitive processes by exciting brain regions involved in working memory and attention using transcranial electrical brain stimulation. Some researchers now believe the cerebellum supports cognition, possibly via a remote neuromodulatory effect on the prefrontal cortex. This paper describes a procedure for investigating a role for the cerebellum in cognition using transcranial direct current stimulation (tDCS), and a selection of information-processing tasks of varying task difficulty, which have previously been shown to involve working memory, attention and cerebellar functioning. One task is called the Paced Auditory Serial Addition Task (PASAT) and the other a novel variant of this task called the Paced Auditory Serial Subtraction Task (PASST). A verb generation task and its two controls (noun and verb reading) were also investigated. All five tasks were performed by three separate groups of participants, before and after the modulation of cortico-cerebellar connectivity using anodal, cathodal or sham tDCS over the right cerebellar cortex. The procedure demonstrates how performance (accuracy, verbal response latency and variability) could be selectively improved after cathodal stimulation, but only during tasks that the participants rated as difficult, and not easy. Performance was unchanged by anodal or sham stimulation. These findings demonstrate a role for the cerebellum in cognition, whereby activity in the left prefrontal cortex is likely dis-inhibited by cathodal tDCS over the right cerebellar cortex. Transcranial brain stimulation is growing in popularity in various labs and clinics. However, the after-effects of tDCS are inconsistent between individuals and not always polarity-specific, and may even be task- or load-specific, all of which requires further study. Future efforts might also be guided towards neuro-enhancement in cerebellar patients presenting with cognitive impairment once a better understanding of brain stimulation mechanisms has emerged.

Transcranial direct current stimulation (tDCS) over the cerebellum exerts a remote effect on the prefrontal cortex, which can modulate cognition and performance. This was demonstrated using two information-processing tasks of varying complexity, whereby only cathodal tDCS improved performance when the task was difficult, but not easy.

调节认知使用小脑经颅直流电刺激

Numerous studies have emerged recently that demonstrate the possibility of modulating, and in some cases enhancing, cognitive processes by exciting brain ...

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 ...

Mapping Inhibitory Neuronal Circuits by Laser Scanning Photostimulation

Inhibitory neurons are crucial to cortical function. They comprise about 20% of the entire cortical neuronal population and can be further subdivided into diverse subtypes based on their immunochemical, morphological, and physiological properties1-4. Although previous research has revealed much about intrinsic properties of individual types of inhibitory neurons, knowledge about their local circuit connections is still relatively limited3,5,6. Given that each individual neuron's function is shaped by its excitatory and inhibitory synaptic input within cortical circuits, we have been using laser scanning photostimulation (LSPS) to map local circuit connections to specific inhibitory cell types. Compared to conventional electrical stimulation or glutamate puff stimulation, LSPS has unique advantages allowing for extensive mapping and quantitative analysis of local functional inputs to individually recorded neurons3,7-9. Laser photostimulation via glutamate uncaging selectively activates neurons perisomatically, without activating axons of passage or distal dendrites, which ensures a sub-laminar mapping resolution. The sensitivity and efficiency of LSPS for mapping inputs from many stimulation sites over a large region are well suited for cortical circuit analysis.
Here we introduce the technique of LSPS combined with whole-cell patch clamping for local inhibitory circuit mapping. Targeted recordings of specific inhibitory cell types are facilitated by use of transgenic mice expressing green fluorescent proteins (GFP) in limited inhibitory neuron populations in the cortex3,10, which enables consistent sampling of the targeted cell types and unambiguous identification of the cell types recorded. As for LSPS mapping, we outline the system instrumentation, describe the experimental procedure and data acquisition, and present examples of circuit mapping in mouse primary somatosensory cortex. As illustrated in our experiments, caged glutamate is activated in a spatially restricted region of the brain slice by UV laser photolysis; simultaneous voltage-clamp recordings allow detection of photostimulation-evoked synaptic responses. Maps of either excitatory or inhibitory synaptic input to the targeted neuron are generated by scanning the laser beam to stimulate hundreds of potential presynaptic sites. Thus, LSPS enables the construction of detailed maps of synaptic inputs impinging onto specific types of inhibitory neurons through repeated experiments. Taken together, the photostimulation-based technique offers neuroscientists a powerful tool for determining the functional organization of local cortical circuits.

This paper introduces an approach of combining laser scanning photostimulation with whole cell recordings in transgenic mice expressing GFP in limited inhibitory neuron populations. The technique allows for extensive mapping and quantitative analysis of local synaptic circuits of specific inhibitory cortical neurons.

测绘激光扫描Photostimulation抑制神经回路

Inhibitory neurons are crucial to cortical function. They comprise about 20% of the entire cortical neuronal population and can be further subdivided into ...

Behavioral Determination of Stimulus Pair Discrimination of Auditory Acoustic and Electrical Stimuli Using a Classical Conditioning and Heart-rate Approach

Acute animal preparations have been used in research prospectively investigating electrode designs and stimulation techniques for integration into neural auditory prostheses, such as auditory brainstem implants1-3 and auditory midbrain implants4,5. While acute experiments can give initial insight to the effectiveness of the implant, testing the chronically implanted and awake animals provides the advantage of examining the psychophysical properties of the sensations induced using implanted devices6,7.
Several techniques such as reward-based operant conditioning6-8, conditioned avoidance9-11, or classical fear conditioning12 have been used to provide behavioral confirmation of detection of a relevant stimulus attribute. Selection of a technique involves balancing aspects including time efficiency (often poor in reward-based approaches), the ability to test a plurality of stimulus attributes simultaneously (limited in conditioned avoidance), and measure reliability of repeated stimuli (a potential constraint when physiological measures are employed).
Here, a classical fear conditioning behavioral method is presented which may be used to simultaneously test both detection of a stimulus, and discrimination between two stimuli. Heart-rate is used as a measure of fear response, which reduces or eliminates the requirement for time-consuming video coding for freeze behaviour or other such measures (although such measures could be included to provide convergent evidence). Animals were conditioned using these techniques in three 2-hour conditioning sessions, each providing 48 stimulus trials. Subsequent 48-trial testing sessions were then used to test for detection of each stimulus in presented pairs, and test discrimination between the member stimuli of each pair. 
This behavioral method is presented in the context of its utilisation in auditory prosthetic research. The implantation of electrocardiogram telemetry devices is shown. Subsequent implantation of brain electrodes into the Cochlear Nucleus, guided by the monitoring of neural responses to acoustic stimuli, and the fixation of the electrode into place for chronic use is likewise shown.

The application of a classical fear conditioning behavioral paradigm for auditory prosthetic research in rats is described. This paradigm provides a mechanism for identifying both detection of, and discrimination between, distinct acoustic and electrical stimuli using heart-rate as an outcome measure.

使用传统的空调和心率的方法刺激听觉声学和电刺激对歧视行为的测定

Acute animal preparations have been used in research prospectively investigating electrode designs and stimulation techniques for integration into neural ...

Transcranial Direct Current Stimulation (tDCS) of Wernicke's and Broca's Areas in Studies of Language Learning and Word Acquisition

Language is a highly important yet poorly understood function of the human brain. While studies of brain activation patterns during language comprehension are abundant, what is often critically missing is causal evidence of brain areas&#39; involvement in a particular linguistic function, not least due to the unique human nature of this ability and a shortage of neurophysiological tools to study causal relationships in the human brain noninvasively. Recent years have seen a rapid rise in the use of transcranial direct current stimulation (tDCS) of the human brain, an easy, inexpensive and safe noninvasive technique that can modulate the state of the stimulated brain area (putatively by shifting excitation/inhibition thresholds), enabling a study of its particular contribution to specific functions. While mostly focusing on motor control, the use of tDCS is becoming more widespread in both basic and clinical research on higher cognitive functions, language included, but the procedures for its application remain variable. Here, we describe the use of tDCS in a psycholinguistic word-learning experiment. We present the techniques and procedures for application of cathodal and anodal stimulation of core language areas of Broca and Wernicke in the left hemisphere of the human brain, describe the procedures of creating balanced sets of psycholinguistic stimuli, a controlled yet naturalistic learning regime, and a comprehensive set of techniques to assess the learning outcomes and tDCS effects. As an example of tDCS application, we show that cathodal stimulation of Wernicke&#39;s area prior to a learning session can impact word learning efficiency. This impact is both present immediately after learning and, importantly, preserved over longer time after the physical effects of stimulation wear off, suggesting that tDCS can have long-term influence on linguistic storage and representations in the human brain.

Transcranial Direct Current Stimulation tDCS of Wernicke's and Broca's Areas in Studies of Language Learning and Word Acquisition

Here, we describe a protocol for using transcranial direct current stimulation for psycho- and neurolinguistic experiments aimed at studying, in a naturalistic yet fully controlled way, the role of cortical areas of the human brain in word learning, and a comprehensive set of behavioral procedures for assessing the outcomes.

韦尼克和布罗卡地区语言学习和文字习得研究中的颅内直接电流刺激(tDCS)

Language is a highly important yet poorly understood function of the human brain. While studies of brain activation patterns during language comprehension ...

Non-Invasive Modulation and Robotic Mapping of Motor Cortex in the Developing Brain

Mapping the motor cortex with transcranial magnetic stimulation (TMS) has potential to interrogate motor cortex physiology and plasticity but carries unique challenges in children. Similarly, transcranial direct current stimulation (tDCS) can improve motor learning in adults but has only recently been applied to children. The use of tDCS and emerging techniques like high&#8211;definition tDCS (HD-tDCS) require special methodological considerations in the developing brain. Robotic TMS motor mapping may confer unique advantages for mapping, particularly in the developing brain. Here, we aim to provide a practical, standardized approach for two integrated methods capable of simultaneously exploring motor cortex modulation and motor maps in children. First, we describe a protocol for robotic TMS motor mapping. Individualized, MRI-navigated 12x12 grids centered on the motor cortex guide a robot to administer single-pulse TMS. Mean motor evoked potential (MEP) amplitudes per grid point are used to generate 3D motor maps of individual hand muscles with outcomes including map area, volume, and center of gravity. Tools to measure safety and tolerability of both methods are also included. Second, we describe the application of both tDCS and HD-tDCS to modulate the motor cortex and motor learning. An experimental training paradigm and sample results are described. These methods will advance the application of non-invasive brain stimulation in children.

We demonstrate protocols for the modulation (tDCS, HD-tDCS) and mapping (robotic TMS) of the motor cortex in children.

发育中大脑运动皮质的非侵入性调制和机器人映射

Mapping the motor cortex with transcranial magnetic stimulation (TMS) has potential to interrogate motor cortex physiology and plasticity but carries ...

Shuttle Box Assay as an Associative Learning Tool for Cognitive Assessment in Learning and Memory Studies using Adult Zebrafish

Cognitive deficits, including impaired learning and memory, are a primary symptom of various developmental and age-related neurodegenerative diseases and traumatic brain injury (TBI). Zebrafish are an important neuroscience model due to their transparency during development and robust regenerative capabilities following neurotrauma. While various cognitive tests exist in zebrafish, most of the cognitive assessments that are rapid examine non-associative learning. At the same time, associative-learning assays often require multiple days or weeks. Here, we describe a rapid associative-learning test that utilizes an adverse stimulus (electric shock) and requires minimal preparation time. The shuttle box assay, presented here, is simple, ideal for novice investigators, and requires minimal equipment. We demonstrate that, following TBI, this shuttle box test reproducibly assesses cognitive deficit and recovery from young to old zebrafish. Additionally, the assay is adaptable to examine either immediate or delayed memory. We demonstrate that both a single TBI and repeated TBI events negatively affect learning and immediate memory but not delayed memory. We, therefore, conclude that the shuttle box assay reproducibly tracks the progression and recovery of cognitive impairment.

Learning and memory are potent metrics in studying either developmental, disease-dependent, or environmentally induced cognitive impairments. Most cognitive assessments require specialized equipment and extensive time commitments. However, the shuttle box assay is an associative learning tool that utilizes a conventional gel box for rapid and reliable assessment of adult zebrafish cognition.

穿梭盒测定作为使用成人斑马鱼进行学习和记忆研究认知评估的关联学习工具

Cognitive deficits, including impaired learning and memory, are a primary symptom of various developmental and age-related neurodegenerative diseases and ...

A Flexible Platform for Monitoring Cerebellum-Dependent Sensory Associative Learning

Climbing fiber inputs to Purkinje cells provide instructive signals critical for cerebellum-dependent associative learning. Studying these signals in head-fixed mice facilitates the use of imaging, electrophysiological, and optogenetic methods. Here, a low-cost behavioral platform (~$1000) was developed that allows tracking of associative learning in head-fixed mice that locomote freely on a running wheel. The platform incorporates two common associative learning paradigms: eyeblink conditioning and delayed tactile startle conditioning. Behavior is tracked using a camera and the wheel movement by a detector. We describe the components and setup and provide a detailed protocol for training and data analysis. This platform allows the incorporation of optogenetic stimulation and fluorescence imaging. The design allows a single host computer to control multiple platforms for training multiple animals simultaneously.

We have developed a single platform to track animal behavior during two climbing fiber-dependent associative learning tasks. The low-cost design allows integration with optogenetic or imaging experiments directed towards climbing fiber-associated cerebellar activity.

用于监测小脑依赖性感觉联想学习的灵活平台

Climbing fiber inputs to Purkinje cells provide instructive signals critical for cerebellum-dependent associative learning. Studying these signals in ...

Assessment of Audio-Tactile Sensory Substitution Training in Participants with Profound Deafness Using the Event-Related Potential Technique

This paper examines the application of electroencephalogram-based methods to assess the effects of audio-tactile substitution training in young, profoundly deaf (PD) participants, with the aim of analyzing the neural mechanisms associated with vibrotactile complex sound discrimination. Electrical brain activity reflects dynamic neural changes, and the temporal precision of event-related potentials (ERPs) has proven to be key in studying time-locked processes while performing behavioral tasks that involve attention and working memory. 
The current protocol was designed to study electrophysiological activity in PD subjects while they performed a continuous performance task (CPT) using complex-sound stimuli, consisting of five different animal sounds delivered through a portable stimulator system worn on the right index finger. As a repeated-measures design, electroencephalogram (EEG) recordings in standard conditions were performed before and after a brief training program (five 1 h sessions over 15 days), followed by offline artifact correction and epoch averaging, to obtain individual and grand-mean waveforms. Behavioral results show a significant improvement in discrimination and a more robust P3-like centroparietal positive waveform for the target stimuli after training. In this protocol, ERPs contribute to the further understanding of learning-related neural changes in PD subjects associated with audio-tactile discrimination of complex sounds.

This protocol is designed to explore underlying learning-related electrophysiological changes in subjects with profound deafness after a short training period in audio-tactile sensory substitution by applying the event-related potential technique.