这项研究是由赠款从德​​国研究基金会（DFG），以MCS（FOR 1581，STU 544/1-1）和OG（FOR 1581，SFB 874）的支持。东风集团的网站是http://www.dfg.de/en/index.jsp。资助者在研究设计，数据收集和分析，发布的决定，或准备manuscript.The作者没有作用感谢托马斯Seidenbecher为我们提供了镀金的协议以及托比亚斯奥托的帮助，建立了电录音设备。

操守准则所有实验均按照动物在科学的保养和使用了德国的指引进行。程序批准了北莱茵 - 威斯特法伦州，德国的状态的国家伦理委员会。 系统概述 操作性试验箱操作性室（ 图1）测量34厘米X 34厘米×50厘米。三个半透明的响应键（4厘米×4厘米位于大约20厘米以上的地面水平）的凹入腔室的后壁上。刺激是通过一台液晶平面显示屏后面装有应答键显示。 2个2瓦的灯泡位于侧壁提供昏暗照明。该室被安置在一个声音衰减柜掩盖多余的声音。扬声器提供白噪声在任何时候。粮食（谷物）是由位于美分下方的食品料斗提供呃键。实验的硬件是由自定义编写的MATLAB代码21控制。动物都通过连接到腔室的前壁数码相机不断监测。 定制的微型硬盘微型硬盘外壳16电极导线定制在我们的实验室;本设计是基于工作由比尔基和同事22,23，和读者可以参考这些文章进行了详细的描述。我们修改了他们的设计，以允许电极的数量较多（16而不是8;25μm的镍铬线），而我们通过导电银胶的探头插座连接的电极布线。此外，我们使用镀金的电极提示，以减少阻抗，以达到更好的信号噪声比（申请-3 V的〜3秒;阻抗应降至&lt;100kΩ的）。 一旦该微驱动器被组装时，电极被切割成所需的长度，提示是CLEANED在超声浴（Tergazyme在蒸馏水中），20分钟，漂洗另一个20分钟在蒸馏水中。镀金的电极尖端应植入前立即生效。对于接地，我们使用一个银球电极置于上方的横向小脑。材料的规范是在材料表，本文附带提供的。 与自由活动动物工作时的一个重要问题是运动伪影。我们发现，在我们设置的运动伪影，主要是由于一）高电极阻抗（&gt; 500KΩ）和插件（植入物）和插座（探头）之间的触点二）不完善的附件，而动物是感动。各种市售microconnectors并没有从自由移动的鸟类记录表现欠佳，因为插头和插座之间的机械接触过鸽子（头部上下摆动，关键啄）蓬勃运动迅速恶化。植入物和探头之间的最佳机械连接​​是由Ginder科学headplug组件来实现。这些插头插座组件配有18接触，并紧紧贴在对方通过环形螺母。 电生理记录设置该电设置包括以下几部分组成：1）一个定制的探头具有单位增益（运算放大器）2）安装在两个机架15差分放大器模块安装装置（DPA-2FS和EPMS-07，分别NPI电子GmbH，德国），3）一个16通道模拟 - 数字转换器（功率1401马克Ⅰ）。原始信号被放大1000倍和带通滤波（500-5000赫兹， 第一阶滤波器），数字化，带有16-20千赫的采样率和存储与Spike2的版本7.06a为脱机处理。是次事件（如刺激发病或动物的个体关键啄）通过实验室构建的并行IO端口盒抓获（见玫瑰; 等人 21），并转发给AD转换器，用于存储沿着与神经生理数据（参见图1）。离线处理在下面进一步描述。 <img alt="图1" fo:content-width="5in" fo:src="/files/ftp_upload/51283/51283fig1highres.jpg" src="/files/ftp_upload/51283/51283fig1.jpg" /> 图1系统概述，信息流是由彩色箭头象征。计算机1控制硬件有关的行为输出（显示刺激通过纯平显示器，房子轻工，食品料斗，振动给料机轻，响应键），并发送事件时间戳的AD转换器。从计算机1接收来自A / D转换器和事件时间戳获得计算机2店神经电生理信号。左边的照片显示的是空调室中的声音衰减柜内。其元素是：1）声音衰减外壳，2-4）响应密钥秒，5）食品料斗，6）馈线光7）房子光，8）观察相机。 单区间强迫选择（SIFC）歧视任务为了清楚起见，我们将在这里介绍的最后SIFC任务，然后解释训练动物在下面这个任务所需的步骤。 该SIFC任务在图2中所概述。后intertrial间隔（ITI）已经过去时，中心键被透照绿色长达5秒（&#39;初始化阶段&#39;）。紧随其后的5秒内动物的第三个反应，一出几个样品的刺激是提出了关于中心键2秒（&#39;样品阶段“;例如刺激是插图所示图2）。后2秒，中心键被再次透照绿色和动物具有前两个侧键透照（&#39;确认相&#39;）响应一次。根据所示的刺激物的身份在采样阶段中，动物被引导到左或右键（“选择相&#39;）的单一反应必需的。如果它选择正确的目标，获得奖励（粒）被授予2秒。因此，任务的核心是由响应左选择键介绍在中心键一个特定的刺激后，并呈现另一种刺激后回应正确的选择键。该样本相位被初始化并确认相包围的一个原因是为了保持动物的头在中心键的前部，而该样品的刺激呈现。 一旦动物主人这一任务的一对刺激（以下简称，“熟悉”的刺激，FS），它提出了一个新颖的刺激（NS）对每一个新的会话，并有学习这两个新异刺激的是然后依次向左侧或右侧的选择键的响应。该FS继续对日期间提交OSE实验作为合适的控制条件。在最后的任务足够的性能关键取决于对动物的意愿来执行&gt; 1,000试验在整体加固概率&lt;0.5。下面的段落描述了一个训练过程，其中任务的复杂性逐渐增加，直到动物到达SIFC的水平;同时，强化概率和每个会话的试验次数需要增加，以确保最后的任务一致的高性能。 1，动物训练<ol><li> 食品限制 <ol><li>至少两周的自由进食后称量动物。就拿这个重量为自由摄食量。限制在未来1-2个星期的食物访问，直到动物达到其自由摄食量的85％。 </li><li>鸽子保持健康的身体外观和正常的活动在整个实验过程中是至关重要的。至为此，仔细留意动物的整个行为训练和测试的整个持续时间外观和重量。前，后各实验阶段，评估每天的食物摄入量称取动物。如果需要提供额外的食物，以防止进一步的减重。提供周末无限制地获得食物。 </li></ol></li><li> Autoshaping Autoshaping用于习惯于对动物的实验室，并建立条件性反应。 <ol><li>目前有5秒的视觉刺激（以下，初始化刺激，IS）的中心键。紧随终止了IS或单个啄来响应键（无论是第一位），关掉按键照明和现在的食物奖励（食品料斗2秒激活）。 </li><li>保持ITI大大长于样品演示时间，以方便学习24。用120秒的值ITI和每天运行40试验。以后重复使用IS在最后的任务初始化，确认和选择关键刺激（ 见图2）。这一阶段的训练将采取动物大约一个星期。 </li><li>一旦动物可靠响应（在试验中&gt; 85％），降低ITI逐步下降到10秒，样品演示时间缩短到2秒。在同一时间，增加所需的加固到3（共3个固定比率，FR 3）的反应的数量。此外，增加每天试验的总数。选择参数，使得该动物被训练每天为约1小时。这一阶段的训练将持续大约2个星期。 </li><li>重复步骤1.2.1 - 1.2.3为左，右按键响应​​，直到受试者可靠地在所有3个按键响应IS。交替试验，激活左，右和中央重点随机的。 </li><li>现在，在中心首先介绍了IS后，有条件的响应，在两边键（省略加固的中心键响应）。备用激活侧键随机试验，以试验。终止试验中，受试者并没有5秒后，中心键响应。重复，直到动物可靠地执行（〜3天）。 </li><li>引进2个新的刺激将在后面作为财政司司长的最后一个任务（参见图2，插图，的例子）。这些刺激1.2.3 - 重复步骤1.2.1。响应会比更迅速地建立了第一个刺激，一般在4天。 </li></ol></li></ol><img alt="图2" fo:content-width="5in" fo:src="/files/ftp_upload/51283/51283fig2highres.jpg" src="/files/ftp_upload/51283/51283fig2.jpg" /> 图2插图行为范式。5秒的ITI后，中心键被透照绿色长达5秒（初始化）。如果动物3X响应这5秒内，1出4;样品的刺激呈现在同一位置。之后的2秒内的动物有回应至少一次固定的样品演示时间，中央啄键时透照再次绿色（确认）。再过啄，2个侧键透照绿色。这个问题表明其选择通过响应一次侧键之一。在定位及重新定位，正确反应之后是2秒粮食获得伴有活化给料机的光，或激活馈线单独的光的。如果不正确，屋里的灯都关闭，持续3秒。在消光，消光刺激正确和不正确的反应仍然微不足道。插图显示的例子熟悉和陌生的刺激对。 <ol start="3"><li> 对于熟悉的刺激训练一个单间隔强迫选择（SIFC）任务 <ol><li>建立初始化，采样，确认和选择的全序列：在每次试验中，目前网络RST了IS（FR 3），那么无论这两个FS（2秒持续时间固定）的，然后再在IS（FR 1）。使用提示选择题的设计：在每次试验中，透照只选择其中的关键是正确的给定的FS。这一阶段的训练大约需要1周。 </li><li>一旦主体可靠地执行（&gt; 85％的反应的各侧键），介绍自由选择试验（在选择阶段透照两个侧面的键）。如果动物响应的正确的一面，提供2秒的粮食获得。不正确的反应都是其次超时处罚（观众席灯光熄灭2秒）。如果没有响应，3秒内发出，终止试用并重新启动ITI。动物平时学习2个星期内的任务，这些子组件。 </li><li>在随后的会话逐渐增加自由选择试验的分数从20％到100％。 </li><li>如果拍摄对象执行&gt;在自由选择试验90％的正确，减少奖励的概率为正确的回应0.5而在平行增加每个会话的试验次数为1,000。不要更改参数天天/会话，但他们选择灵活根据有关初始化遗漏，并正确反应百分比主体的性能水平。这一阶段的训练将持续约4个星期。 </li><li>鸽子往往拒绝回应不熟悉的刺激。因此，一旦动物能够可靠地执行&gt; 1,000试验，自选图形应对大集视觉刺激（见1.2节）。但是，不要preexpose注定供以后使用作为最后的范式，但彩色显示器均匀新颖刺激的视觉刺激。 </li></ol></li><li> 最后的单间隔强迫选择任务与新刺激下不同加固条件 <ol><li>热身 让受试者进行50次试验，只有在FS。设置奖励的概率为这些刺激，以&lt;1（例如，0.5 - 0.8）在所有阶段，以防止过早饱食和，因此，缺乏动力响应的。 </li><li>收购阶段 随机交替试验与演示FS和生理盐水。分配不同的反应键作为正确的两个NS和强化每一个正确的回应。计算正确反应百分比的运行平均值在过去的120试验。收购被视为完整的一次为每个对NS的性能超过85％，但不是最低的150试验之前已经执行。 </li><li>消光舞台 停止补强正确和惩治不正确的反应，以一个随机的NS（消光刺激）。开始重新捕获阶段时，正确的响应灭绝的刺激降到低于60％，动物至少经历了150试验在这个阶段中总。 </li><li>重新捕获阶段 再次加强正确和惩罚不正确的反应，以消光刺激，如在收购阶段。终止会话时，这个刺激的表现超过85％和阿尼玛LS共进行至少150试验在这个阶段。 </li></ol></li></ol> 2，电<ol><li> 电极植入 植入手术发生动物后反复（3-4倍）完成整个采集灭绝，再获取序列并进行更详细的别处25所述。 <ol><li>将5-6不锈钢microsrews头骨上的锚定牙科水泥头安装，包括微硬盘。 </li><li>进行开颅手术略高于感兴趣的大脑区域;然后小心地解剖硬脑膜和降低电极至所需的位置。 </li><li>前锚定微硬盘的磁头安装，应用凡士林围绕引导套管;这将防止牙科用粘固剂从包围所述导向管。 </li><li>使用绝缘银球电极放置在颅骨上覆小脑地面之下。 </li><li>提供动物的止痛药（卡洛芬，10毫克/ K克，注射，每天两次）三天手术。让动物恢复至少2周。 </li></ol></li><li> 录音虽然动物执行任务 <ol><li>在开始之前，请使用新的对新刺激的每个会话，并提前电极至少125微米（驱动螺杆半圈）。如果有足够的信号噪声比没有动作电位的观察，中止会话，将动物的笼子，然后重试的第二天。 </li><li>排列的探头电缆，使得它不与动物的正常啄食和摄食行为干扰。这可以通过附加的连接线与多个弹性带的空调室的顶部和鸟类habituating到连接电缆一段时间来实现。 </li><li>如果有的话，做一个换向器，为鸟类提供了运动的额外自由使用。 </li></ol></li><li> 离线信号分析 <ol><li>带-P屁股过滤器由500至5,000 Hz使用Spike2的陡峭滚降下线的所有通道。提取尖峰幅度阈值并进行排序采用手动主成分分析。 </li><li>检查结果排序与自定义编写的MATLAB代码（可在MATLAB中心文件交换，文件编号＃37339）。良好的隔离单单元（ 图3）应符合以下所有条件：1）在主成分空间明显分开集群，多台机组二）没有任何迹象时，所有记录的波形叠加并绘制热图（ 图3A ），C）对称分布的峰值波形振幅（ 图3B），在整个会议d）稳定的记录就证明了不变的尖峰幅度（ 图3C），e）在前面的秒杀的不应期发生的没有或很少有秒杀活动（ 图3D），且至少2（SNR的信号-噪声比（SNR）是这里ðefined的平均尖峰波形，由噪声频带（2.5 个和值的所有波形的第一个bin中的分布为97.5 个百分位数））的修剪宽度除以最小值和最大值之间的差异。在图3所示的单元的SNR为3.9。 </li><li>原道脱机运动有关的文物考察。当表示放弃渠道。 </li><li>在关键啄发生电气构件都可以在极少数情况下被混淆与适当的尖峰波形。测试的录音通过检查尖峰时间直方图污染计算相对于每个注册的关键啄（围啄时间直方图，PPTH， 图3E）。啄引起的伪影显示为直方图接近峰值（±50毫秒），以时间为0。作为一个额外的检查，绘制在±20毫秒的关键啄的注册分别假定所有秒杀活动的波形，并比较它秒杀波形检测outsidË这个窗口（ 图3F）。 </li></ol></li></ol><img alt="图3" fo:content-width="4in" src="/files/ftp_upload/51283/51283fig3highres.jpg" width="400" /> 图3。质量度量单元隔离。 A）热，所有波形的时间-电压值B）的最大值（红色），最小值（绿色），和所有的噪声波形（蓝色）电压值的分布地图。该分布很好的分离，这表明优异的设备隔离。C）放电频率（红色，从2秒片段中的所有intertrial间隔计算）和尖峰振幅（峰-峰值）作为时间的会话函数。这两个曲线平滑与棚车功能（宽度：50个数据点）D）峰峰间隔分布的本机。滨宽，10毫秒（插图：1毫秒）。很短的时间间隔几乎不存在（&lt;下4毫秒的时间间隔为0.1％）。 &lt;STRONG&gt; E）PSTH触发键啄。接近关键啄（±20毫秒）事件计数红色突出显示。 女性）在±20毫秒啄关键事件的记录全部157波形。波形媲美面板中A所示整体的波形形状

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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Components+of+Tone-Evoked+Spike+Trains+in+Auditory+Cortex+and+Lateral+Amygdala.">Components of Tone-Evoked Spike Trains in Auditory Cortex and Lateral Amygdala.</a> Neuron. 19, 613-624 (1997).</li><li>Maren, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auditory+fear+conditioning+increases+CS-elicited+spike+firing+in+lateral+amygdala+neurons+even+after+extensive+overtraining.">Auditory fear conditioning increases CS-elicited spike firing in lateral amygdala neurons even after extensive overtraining.</a> Eur J Neurosci. 12, 4047-4054 (2000).</li><li>Milad, M. R., Quirk, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurons+in+medial+prefrontal+cortex+signal+memory+for+fear+extinction.">Neurons in medial prefrontal cortex signal memory for fear extinction.</a> Nature. 420, 713-717 (2002).</li><li>Hobin, J. A., Goosens, K. A., Maren, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Context-dependent+neuronal+activity+in+the+lateral+amygdala+represents+fear+memories+after+extinction.">Context-dependent neuronal activity in the lateral amygdala represents fear memories after extinction.</a> J Neurosci. 23, 8410-8416 (2003).</li><li>Maren, S., Hobin, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hippocampal+regulation+of+context-dependent+neuronal+activity+in+the+lateral+amygdala.">Hippocampal regulation of context-dependent neuronal activity in the lateral amygdala.</a> Learn Mem. 14, 318-324 (2007).</li><li>Knapska, E., 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=Differential+involvement+of+the+central+amygdala+in+appetitive+versus+aversive+learning.">Differential involvement of the central amygdala in appetitive versus aversive learning.</a> Learn Mem. 13, 192-200 (2006).</li><li>Harloe, J. P., Thorpe, A. J., Lichtman, A. 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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relative+and+absolute+strength+of+response+as+a+function+of+frequency+of+reinforcement.">Relative and absolute strength of response as a function of frequency of reinforcement.</a> J Exp Anal Behav. 4, 267-272 (1961).</li><li>Brown, P. L., Jenkins, H. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auto-shaping+of+the+pigeon's+key-peck.">Auto-shaping of the pigeon's key-peck.</a> J Exp Anal Behav. 11, 1-8 (1968).</li><li>Epstein, R., Kirshnit, C. E., Lanza, R. P., Rubin, L. 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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complex+visual+concept+in+the+pigeon.">Complex visual concept in the pigeon.</a> Science. 146, 549-551 (1964).</li><li>St&#252;ttgen, M. C., Yildiz, A., G&#252;nt&#252;rk&#252;n, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adaptive+criterion+setting+in+perceptual+decision+making.">Adaptive criterion setting in perceptual decision making.</a> J Exp Anal Behav. 96, 155-176 (2011).</li><li>St&#252;ttgen, M. C., Kasties, N., Lengersdorf, D., Starosta, S., G&#252;nt&#252;rk&#252;n, O., J&#228;kel, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suboptimal+criterion+setting+in+a+perceptual+choice+task+with+asymmetric+reinforcement.">Suboptimal criterion setting in a perceptual choice task with asymmetric reinforcement.</a> Behav Processes. 96, 59-70 (2013).</li><li>Rose, J., Otto, T., Dittrich, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Biopsychology-Toolbox:+a+free,+open-source+Matlab-toolbox+for+the+control+of+behavioral+experiments.">The Biopsychology-Toolbox: a free, open-source Matlab-toolbox for the control of behavioral experiments.</a> J Neurosci Methods. 175, (2008).</li><li>Bilkey, D. K., Muir, G. 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作者什么都没有透露。

本协议描述适用于并发单机录制一个复杂的行为的任务。我们所描述的SIFC任务的鸽子，但它可以通过要求的鼻子戳或杆按下，而不是关键啄，而代以通过视觉嗅觉，听觉或触觉的刺激很容易地适应啮齿类动物。 也许是训练过程中最关键的步骤是：1）逐渐减少奖励概率和2）在试验的数目增加。对于间歇性强化的熟悉的刺激，我们决定奖励的概率从0.5到0.8;这些都足以产生稳定?...

学习新的刺激 - 反应 - 结果组织从事广泛的神经可塑性过程。这些过程最终体现在单个神经元的变化尖峰输出。可以说，最经常使用的学习范式之一是与啮齿类动物进行巴甫洛夫恐惧制约。在这种背景下，一个条件反应的采集和灭绝采取了几十个试验2内进行。条件性恐惧的快速发展是有利的，因为它允许在短时间内运行大量的动物。此外，收购和消光可以在几十试验在一天中天真的动物观察到3,4或分布在2〜3天2,5-8。然而，见解学习期间在这些实验中并不一定适用恐惧条件外域上涨约神经活动的变化。例如，目标导向的行为由驱动位置略去钢筋更充分地通过操作性，而不是巴甫洛夫条件反射过程建模，并可能部分依赖于不同的神经基础9,10。此外，恐惧制约发展如此迅速，在CS的神经反应只能为几十个试验中观察到，将神经活动的变化分析严重限制在学习。 不幸的是，操作性通常响应的收购和消灭需要很多天。这是有害的神经生理学研究，因为它是非常困难的在超过几小时来记录单细胞的活性。由于细胞外记录的动作电位波形的高相似性，它要求该记录在一天尖峰是从同一小区产生的尖峰与记录在下一11,12相似的波形，特别是在具有高的区域是有问题细胞密度如海马。 哥伦比亚利维亚经家蝇 ）是经典模式生物在实验心理学13-17。这些鸟能够执行复杂的视觉歧视18，可以灵活适应行为来改变筋突发事件19,20，并且是唯一热衷的工人，进行1,000试验的构件最小配筋量。这些特性使它们特别适用于下述实验。

<table border="0" cellpadding="0" cellspacing="0" style="width:1379px;" width="1377">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:277px;">Name of Material/ Equipment</td>
			<td style="width:305px;">Company</td>
			<td style="width:325px;">Catalog Number</td>
			<td style="width:471px;">Comments/Description</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:277px;">Resistance wire (for use as electrodes)</td>
			<td style="width:305px;">California Fine Wire, Grover Beach (CA), USA</td>
			<td></td>
			<td>Stablohm 675; formvar-coated nichrome wires (outer diameter 25 &#181;m)</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:277px;">Microconnectors</td>
			<td style="width:305px;">Ginder Scientific, Nepean, Ontario, Canada</td>
			<td style="width:325px;"></td>
			<td>GS18PLG-220 (plug) &#38; GS18SKT-220 (socket to build headstage)</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:277px;">Cannulae</td>
			<td style="width:305px;">Henke Sass Wolf, Tuttlingen, Germany</td>
			<td style="width:325px;"></td>
			<td>0.4x20mm/ 27Gx3/4&#34;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:277px;">Gold solution for plating</td>
			<td style="width:305px;">Neuralynx, Bozeman (MT), USA</td>
			<td style="width:325px;"></td>
			<td>SIFCO Process Gold Non-Cyanide, Code 5355</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:277px;">Solution for ultrasonic bath</td>
			<td style="width:305px;">Alconox, Inc., New York, USA</td>
			<td style="width:325px;">1304</td>
			<td>Tergazyme</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:277px;">Conductive glue</td>
			<td style="width:305px;">Henkel Loctite</td>
			<td></td>
			<td>LOCTITE 3888 Silver filled, conductive, adhesive</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:277px;">Stainless steel screws</td>
			<td style="width:305px;">J.I. Morris, Southbridge (MA), USA</td>
			<td style="width:325px;"></td>
			<td>F0CE125 self-tapping miniature screws, body length 1/8 inches</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:277px;">Light-curing dental cement</td>
			<td style="width:305px;">van der Ven Dental, Duisburg, Germany</td>
			<td style="width:325px;"></td>
			<td>Omniceram Evo Flow A2</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:277px;">Light-curing unit</td>
			<td style="width:305px;">van der Ven Dental, Duisburg, Germany</td>
			<td style="width:325px;"></td>
			<td>Jovident Excelled 215 Curing Light (wireless LED light curing unit)</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:277px;">Filter amplifiers</td>
			<td style="width:305px;">npi electronic GmbH, Germany</td>
			<td>DPA-2FS</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:277px;">A/D converter</td>
			<td style="width:305px;">Cambridge Electronic Design, Cambridge, UK</td>
			<td>power 1401</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:277px;">Spike2 software</td>
			<td style="width:305px;">Cambridge Electronic Design, Cambridge, UK</td>
			<td style="width:325px;"></td>
			<td>Version 7.06a</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:277px;">Matlab</td>
			<td style="width:305px;">The Mathworks, Natick (MA), USA</td>
			<td style="width:325px;"></td>
			<td>R2012a</td>
		</tr>
	</tbody>
</table>

recording single neurons' action potentials from freely moving pigeons across three stages of learning

而学习的主题，吸引了极大的兴趣来自行为和神经科学家，只有相对少数的研究者观察到单个神经元的活动，而动物被收购的operantly条件反应，或当该响应熄灭。但即使在这种情况下，观察期通常包括学习的只有一个阶段， 即收购或灭绝，但不能同时使用（例外情况包括用人逆转的学习方案;见宾曼等 1中的示例）。然而，收购和灭绝涉及不同的学习机制，因此预期将伴随着不同类型和/或神经可塑性的基因位点。 因此，我们开发了一种行为范式的机构学在一个单一的行为会话和其中三个阶段是非常适合用于同时记录的单个神经元的动作电位。动物重新训练上的单间隔强迫选择的任务，需要各两个可能的选择反应，不同的新颖的视觉刺激（购入）的呈现映射。已达到预定的性能指标，这两个选择反应之一后不再增强（消光）。按照一定的递减在性能水平，正确的反应是再次强化（重捕）。通过使用一套新的在每一个会话的刺激，动物可以反复进行收购灭绝 - 再获取过程。因为学习的三个阶段发生在一个单一的行为会议上，范式是理想的同时观测多个单个神经元的扣球输出。我们用鸽子作为模型系统，但是该任务可以很容易地适应空调能力辨别学习的任何其它物种。

行为 图4A显示了一个动物在一个实例中会话的行为表现。动物的表现水平达到标准的NS 2在180个试验（45刺激演示），并接近100％的NS从1开始。这一策略 - 首先响应相同的密钥进行新的刺激，然后调整反应的刺激之一 - 大约是经常观察到的初始随机响应两个NS。在这个会议上，南北2是随机选择接受灭绝，这意味着下面的这种刺激所有的选择保持无关紧要的（学习阶段?...

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Recording Single Neurons&#039; Action Potentials from Freely Moving Pigeons Across Three Stages of Learning

学习新的刺激 - 反应协会从事范围广泛的神经工序，而最终体现在改变单个神经元的秒杀输出。在这里，我们描述一个行为协议，允许单神经元活动的连续记录，而动物收购，熄灭，和一个实验性会话中重新获取一个条件反应。

自由移动鸽子穿越三个阶段学习记录单个神经元&#39;动作电位

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed ...

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Recording Single Neurons' Action Potentials from Freely Moving Pigeons Across Three Stages of Learning

11.9K 次观看.  Ruhr-University Bochum. 学习新的刺激 - 反应协会从事范围广泛的神经工序，而最终体现在改变单个神经元的秒杀输出。在这里，我们描述一个行为协议，允许单神经元活动的连续记录，而动物收购，熄灭，和一个实验性会话中重新获取一个条件反应。 

视频: Recording Single Neurons' Action Potentials from Freely Moving Pigeons Across Three Stages of Learning

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from individual neurons are critical for understanding how neural circuits function under natural conditions. Traditionally, these recordings have been performed 'blind', meaning the identity of the recorded cell is unknown at the start of the recording. Cellular identity can be subsequently determined via intracellular1, juxtacellular2 or loose-patch3 iontophoresis of dye, but these recordings cannot be pre-targeted to specific neurons in regions with functionally heterogeneous cell types. Fluorescent proteins can be expressed in a cell-type specific manner permitting visually-guided single-cell electrophysiology4-6. However, there are many model systems for which these genetic tools are not available. Even in genetically accessible model systems, the desired promoter may be unknown or genetically homogenous neurons may have varying projection patterns. Similarly, viral vectors have been used to label specific subgroups of projection neurons7, but use of this method is limited by toxicity and lack of trans-synaptic specificity. Thus, additional techniques that offer specific pre-visualization to record from identified single neurons in vivo are needed. Pre-visualization of the target neuron is particularly useful for challenging recording conditions, for which classical single-cell recordings are often prohibitively difficult8-11. The novel technique described in this paper uses retrograde transport of a fluorescent dye applied using tungsten needles to rapidly and selectively label a specific subset of cells within a particular brain region based on their unique axonal projections, thereby providing a visual cue to obtain targeted electrophysiological recordings from identified neurons in an intact circuit within a vertebrate CNS.
The most significant novel advancement of our method is the use of fluorescent labeling to target specific cell types in a non-genetically accessible model system. Weakly electric fish are an excellent model system for studying neural circuits in awake, behaving animals12. We utilized this technique to study sensory processing by "small cells" in the anterior exterolateral nucleus (ELa) of weakly electric mormyrid fish. "Small cells" are hypothesized to be time comparator neurons important for detecting submillisecond differences in the arrival times of presynaptic spikes13. However, anatomical features such as dense myelin, engulfing synapses, and small cell bodies have made it extremely difficult to record from these cells using traditional methods11, 14. Here we demonstrate that our novel method selectively labels these cells in 28% of preparations, allowing for reliable, robust recordings and characterization of responses to electrosensory stimulation.

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde transport of fluorescent dye labels a sub-population of neurons based on anatomical projection. Labeled axons can be visually targeted in vivo, permitting extracellular recording from identified axons. This technique facilitates recording when neurons cannot be labeled through genetic manipulation or are difficult to isolate using 'blind' in vivo approaches.

逆行荧光染料标记允许有针对性的外单细胞记录技术鉴定神经元<em&gt;在体内</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

科研

神经科学

Optical Recording of Suprathreshold Neural Activity with Single-cell and Single-spike Resolution

Signaling of information in the vertebrate central nervous system is often carried by populations of neurons rather than individual neurons. Also propagation of suprathreshold spiking activity involves populations of neurons. Empirical studies addressing cortical function directly thus require recordings from populations of neurons with high resolution. Here we describe an optical method and a deconvolution algorithm to record neural activity from up to 100 neurons with single-cell and single-spike resolution. This method relies on detection of the transient increases in intracellular somatic calcium concentration associated with suprathreshold electrical spikes (action potentials) in cortical neurons. High temporal resolution of the optical recordings is achieved by a fast random-access scanning technique using acousto-optical deflectors (AODs)1. Two-photon excitation of the calcium-sensitive dye results in high spatial resolution in opaque brain tissue2. Reconstruction of spikes from the fluorescence calcium recordings is achieved by a maximum-likelihood method. Simultaneous electrophysiological and optical recordings indicate that our method reliably detects spikes (&gt;97% spike detection efficiency), has a low rate of false positive spike detection (&lt; 0.003 spikes/sec), and a high temporal precision (about 3 msec) 3. This optical method of spike detection can be used to record neural activity in vitro and in anesthetized animals in vivo3,4.

Understanding the function of the vertebrate central nervous system requires recordings from many neurons because cortical function arises on the level of populations of neurons. Here we describe an optical method to record suprathreshold neural activity with single-cell and single-spike resolution, dithered random-access scanning. This method records somatic fluorescence calcium signals from up to 100 neurons with high temporal resolution. A maximum-likelihood algorithm deconvolves the underlying suprathreshold neural activity from the somatic fluorescence calcium signals. This method reliably detects spikes with high detection efficiency and a low rate of false positives and can be used to study neural populations in vitro and in vivo.

阈上的神经活动的单细胞和单穗的分辨率的光学的方法记录

Signaling of information in the vertebrate central nervous system is often carried by populations of neurons rather than individual neurons. Also ...

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such as epilepsy and neurodegenerative disorders. However, it is technically difficult to obtain EEG recordings in neonates as it requires specialized handling and great care. Here, we present a novel method to record EEG in neonatal rat pups (P8-P15). We designed a simple and reliable electrode using computer pin loci; it can be easily implanted into the skull of a rat pup to record high-quality EEG signals in the normal and epileptic brain. Pups were given an intraperitoneal (i.p.) injection of the neurotoxin kainic acid (KA) to induce epileptic seizures. The surgical implantation performed in this procedure is less expensive than other EEG procedures for neonates. This method allows one to record high-quality and stable EEG signals for more than 1 week. Furthermore, this procedure can also be applied to adult rats and mice to study epilepsy or other neurological disorders.

Here, we introduce a novel technique designed to record electroencephalography (EEG) in freely moving neonatal epileptic pups and describe its procedures, features, and applications. This method allows one to record EEG for more than 1 week.

使用新颖的方法在自由移动的新生大鼠中记录脑电图

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such ...

Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls body functions and internal organ rhythms when animals are exposed to emotional challenges and changes in their living environments. In general experiments, signals from different organs, such as the brain and the heart, are recorded by independent recording systems that require multiple recording devices and different procedures for processing the data files. This study describes a new method that can simultaneously monitor electrical biosignals, including tens of local field potentials in multiple brain regions, electrocardiograms that represent the cardiac rhythm, electromyograms that represent awake/sleep-related muscle contraction, and breathing signals, in a freely moving rat. The recording configuration of this method is based on a conventional micro-drive array for cortical local field potential recordings in which tens of electrodes are accommodated, and the signals obtained from these electrodes are integrated into a single electrical board mounted on the animal's head. Here, this recording system was improved so that signals from the peripheral organs are also transferred to an electrical interface board. In a single surgery, electrodes are first separately implanted into the appropriate body parts and the target brain areas. The open ends of all of these electrodes are then soldered to individual channels of the electrical board above the animal's head so that all of the signals can be integrated into the single electrical board. Connecting this board to a recording device allows for the collection of all of the signals into a single device, which reduces experimental costs and simplifies data processing, because all data can be handled in the same data file. This technique will aid the understanding of the neurophysiological correlates of the associations between central and peripheral organs.

This study introduces a method for the simultaneous recording of local field potentials in the brain, electrocardiograms, electromyograms, and breathing signals of a freely moving rat. This technique, which reduces experimental costs and simplifies data analysis, will contribute to the understanding of the interactions between the brain and peripheral organs.

自由运动大鼠皮质局部电位、心电图、肌电图和呼吸节律的同步记录

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls ...

Noninvasive EEG Recordings from Freely Moving Piglets

The method allows the recording of high-quality electroencephalograms (EEGs) from freely moving piglets directly in the pigpen. We use a one-channel telemetric electroencephalography system in combination with standard self-adhesive hydrogel electrodes. The piglets are calmed down without the use of sedatives. After their release into the pigpen, the piglets behave normally&#8212;they drink and sleep in the same cycle as their siblings. Their sleep phases are used for the EEG recordings.

Here, we present a protocol to record telemetric electroencephalograms (EEGs) from freely moving piglets directly in the pigpen without the use of a sedative, making it possible to record typical EEG patterns during non-REM sleep, like spindle bursts.

自由移动仔猪的无创脑电图记录

The method allows the recording of high-quality electroencephalograms (EEGs) from freely moving piglets directly in the pigpen. We use a one-channel ...

Microdialysis of Excitatory Amino Acids During EEG Recordings in Freely Moving Rats

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain interstitial space with the behavior and/or with the specific outcome of a pathology (e.g., seizures for epilepsy). When studying epilepsy, the microdialysis technique is often combined with short-term or even long-term video-electroencephalography (EEG) monitoring to assess spontaneous seizure frequency, severity, progression and clustering. The combined microdialysis-EEG is based on the use of several methods and instruments. Here, we performed in vivo microdialysis and continuous video-EEG recording to monitor glutamate and aspartate outflow over time, in different phases of the natural history of epilepsy in a rat model. This combined approach allows the pairing of changes in the neurotransmitter release with specific stages of the disease development and progression. The amino acid concentration in the dialysate was determined by liquid chromatography. Here, we describe the methods and outline the principal precautionary measures one should take during in vivo microdialysis-EEG, with particular attention to the stereotaxic surgery, basal and high potassium stimulation during microdialysis, depth electrode EEG recording and high-performance liquid chromatography analysis of aspartate and glutamate in the dialysate. This approach may be adapted to test a variety of drug or disease induced changes of the physiological concentrations of aspartate and glutamate in the brain. Depending on the availability of an appropriate analytical assay, it may be further used to test different soluble molecules when employing EEG recording at the same time.

Here, we describe a method for in vivo microdialysis to analyze aspartate and glutamate release in the ventral hippocampus of epileptic and non-epileptic rats, in combination with EEG recordings. Extracellular concentrations of aspartate and glutamate may be correlated with the different phases of the disease.

自由运动大鼠脑电图记录过程中兴奋氨基酸的微透析

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain ...

Hybrid Microdrive System with Recoverable Opto-Silicon Probe and Tetrode for Dual-Site High Density Recording in Freely Moving Mice

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, conventional microdrive designs often only allow use of one type of electrode to record from single or multiple regions, limiting the yield of single-unit or depth profile recordings. It also often limits the ability to combine electrode recordings with optogenetic tools to target pathway and/or cell type specific activity. Presented here is a hybrid microdrive array for freely moving mice to optimize yield and a description of its fabrication and reuse of the microdrive array. The current design employs nine tetrodes and one opto-silicon probe implanted in two different brain areas simultaneously in freely moving mice. The tetrodes and the opto-silicon probe are independently adjustable along the dorsoventral axis in the brain to maximize the yield of unit and oscillatory activities. This microdrive array also incorporates a set-up for light, mediating optogenetic manipulation to investigate the regional- or cell type-specific responses and functions of long-range neural circuits. In addition, the opto-silicon probe can be safely recovered and reused after each experiment. Because the microdrive array consists of 3D-printed parts, the design of microdrives can be easily modified to accommodate various settings. First described is the design of the microdrive array and how to attach the optical fiber to a silicon probe for optogenetics experiments, followed by fabrication of the tetrode bundle and implantation of the array into a mouse brain. The recording of local field potentials and unit spiking combined with optogenetic stimulation also demonstrate feasibility of the microdrive array system in freely moving mice.

This protocol describes the construction of a hybrid microdrive array that allows implantation of nine independently adjustable tetrodes and one adjustable opto-silicon probe in two brain regions in freely moving mice. Also demonstrated is a method for safely recovering and reusing the opto-silicon probe for multiple purposes.

混合微驱系统,带可回收的光电探头和Tetrode,用于自由移动小鼠的双位点高密度记录

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, ...

Multi-Fiber Photometry to Record Neural Activity in Freely-Moving Animals

Recording the activity of a group of neurons in a freely-moving animal is a challenging undertaking. Moreover, as the brain is dissected into smaller and smaller functional subgroups, it becomes paramount to record from projections and/or genetically-defined subpopulations of neurons. Fiber photometry is an accessible and powerful approach that can overcome these challenges. By combining optical and genetic methodologies, neural activity can be measured in deep brain structures by expressing genetically-encoded calcium indicators, which translate neural activity into an optical signal that can be easily measured. The current protocol details the components of a multi-fiber photometry system, how to access deep brain structures to deliver and collect light, a method to account for motion artifacts, and how to process and analyze fluorescent signals. The protocol details experimental considerations when performing single and dual color imaging, from either single or multiple implanted optic fibers.

This protocol details how to implement and perform multi-fiber photometry recordings, how to correct for calcium-independent artifacts, and important considerations for dual-color photometry imaging.

多纤维光测量记录自由移动动物的神经活动

Recording the activity of a group of neurons in a freely-moving animal is a challenging undertaking. Moreover, as the brain is dissected into smaller and ...

Wireless Electrophysiological Recording of Neurons by Movable Tetrodes in Freely Swimming Fish

The neural mechanisms governing fish behavior remain mostly unknown, although fish constitute the majority of all vertebrates. The ability to record brain activity from freely moving fish would advance research on the neural basis of fish behavior considerably. Moreover, precise control of the recording location in the brain is critical to studying coordinated neural activity across regions in fish brain. Here, we present a technique that records wirelessly from the brain of freely swimming fish while controlling for the depth of the recording location. The system is based on a neural logger associated with a novel water-compatible implant that can adjust the recording location by microdrive-controlled tetrodes. The capabilities of the system are illustrated through recordings from the telencephalon of goldfish.

A novel wireless technique for recording extracellular neural signals from the brain of freely swimming goldfish is presented. The recording device is composed of two tetrodes, a microdrive, a neural data logger, and a waterproof case. All parts are custom-made except for the data logger and its connector.

自由游泳鱼中可移动的特龙对神经元的无线电生理记录

The neural mechanisms governing fish behavior remain mostly unknown, although fish constitute the majority of all vertebrates. The ability to record brain ...

Single-Cell Electroporation across Different Organotypic Slice Culture of Mouse Hippocampal Excitatory and Class-Specific Inhibitory Neurons

Electroporation has established itself as a critical method for transferring specific genes into cells to understand their function. Here, we describe a single-cell electroporation technique that maximizes the efficiency (~80%) of in vitro gene transfection in excitatory and class-specific inhibitory neurons in mouse organotypic hippocampal slice culture. Using large glass electrodes, tetrodotoxin-containing artificial cerebrospinal fluid and mild electrical pulses, we delivered a gene of interest into cultured hippocampal CA1 pyramidal neurons and inhibitory interneurons. Moreover, electroporation could be carried out in cultured hippocampal slices up to 21 days in vitro with no reduction in transfection efficiency, allowing for the study of varying slice culture developmental stages. With interest growing in examining the molecular functions of genes across a diverse range of cell types, our method demonstrates a reliable and straightforward approach to in vitro gene transfection in mouse brain tissue that can be performed with existing electrophysiology equipment and techniques.

Presented here is a protocol for single-cell electroporation that can deliver genes in both excitatory and inhibitory neurons across a range of in vitro hippocampal slice culture ages. Our approach provides precise and efficient expression of genes in individual cells, which can be used to examine cell-autonomous and intercellular functions.

小鼠河马兴奋和类特异性抑制神经元的不同组织型切片培养的单细胞电位

Electroporation has established itself as a critical method for transferring specific genes into cells to understand their function. Here, we describe a ...