一些这里显示的数据的获得在杰奎琳全克劳利博士在心理健康的美国国家研究所的实验室，我们要感谢她让我们展示的文件中的数据。我们也感谢中西一夫，他在制定行为分析ImageFZ程序的帮助。这项研究得到了支持格兰特 - 基盘研究（B）（21300121），格兰特在急救科学研究的创新领域（综合脑科学网）由教育部，科学，体育和文化日本，授予来自日本神经信息学中心（NIJC），并从日本科学技术振兴机构（JST）的CREST补助。

所有的实验中，应根据当地动物护理和使用委员会制定的指导和协议来执行。 1。设备设置<ol><li>用于调节和上下文试验装置是一个方腔室与electrifiable格栅地板，声源，和一个校准震动发生器。各种房型大小时，从54厘米尺寸不同×27厘米×30厘米21到25公分X 35公分x30厘米22。在此协议中，该装置包括一个丙烯酸类正方形室（33厘米×25厘米×28厘米;透明的前壁和后壁;白在侧壁）与金属网格（0.2cm直径的，间隔开的0.5厘米分开）覆盖通过透明的丙烯酸盖（ 图1A）。有必要将室内的白色亚克力层（ 图1B）来分析黑，刺鼠，或稀棕色小鼠的行为，因为图像分析系统与ImageFZ软件程序（可免费下载，见表材料/试剂）区别于白色背景中的每个捕获的视频图像较暗的对象。白化小鼠也可以使用黑色的金属网格和黑色亚克力层（ 图1B）进行测试。发光二极管（LED）灯被安装在装置上方的天花板上。网格地板是由LED灯点亮，在100勒克司。连接到一个白噪声/音调发生器（ 图1C）扬声器安装在天花板上5厘米上面的盖子呈现听觉提示（白噪声55 DB）作为条件刺激（CS）。该网格连接到一个震动发生器（ 图1C），可提供电动电击作为非条件刺激（美国）。试验室被放置在一个隔音房间（170厘米×210厘米×200厘米）（ 图1D）在测试期间尽量减少外部噪声。这个条件也防止了没有被经受的小鼠从听觉的听觉线索或测试小鼠的发声测试。 </li><li>对于提示的测试装置被构成的，即具有从调节室不同的性质的腔室，提供了新的上下文。当务之急是要改变感官线索尽可能使鼠标感知新颖的历境作为是无关的调节室。一般地，一个不同形状的方块或三角形腔室被使用。此外，不同的照明和/或嗅觉也提供到鼠标。在这个协议中，该装置是一种丙烯酸三角形腔室（33厘米×29厘米×32厘米;白在每个侧壁）与平坦，白色的地板为黑色，刺豚鼠，或稀棕色小鼠或平坦，黑色地板白化小鼠，覆盖着透明的压克力盖子（ 图1E）。 LED灯安装在装置上方的天花板上。地板的照度水平被设定为30勒克司。一位发言者是安装在天花板上5厘米上面的盖子公关ESENT听觉提示说是一样的，在空调的时间提供给小鼠。三角室坐落在一个不同的隔音室从其中空调和上下文测试执行了房间。 </li><li>每个腔室都配备了安装在天花板上电荷耦合器件（CCD）摄像机通过四通道视频分配器和USB影像撷取装置来监视鼠标的行为，以及设备和鼠标的图像连接到Windows电脑被捕获和分析的应用软件程序ImageFZ（见协议6）。白噪声和电击发生器是由ImageFZ软件程序自动控制，开始时间和白噪声和电击的持续时间必须被写入到一个文本文件（见的视频显示一个示例文本文件&#39;简单COND“关于如何将参数写入到文件中的信息），该被读入的应用程序。 </li><li>在每次测试开始时，丙烯酸墙壁和地板擦拭浸泡在超级次氯酸水（pH 6-7）毛巾，以及网格用70％乙醇清洗，以防止基于嗅觉的偏压。的网格，用乙醇擦拭，而不是超次氯酸水，以确保该网格并不减少其电导率由于生锈。 </li></ol> 2。动物的制备<ol><li>一般来说，两到四个小鼠在温度控制持房（23±2℃）与12小时亮/暗周期（ 如在早上7:00开灯）住每笼。 </li><li>在这个协议中，以减少运输笼的可能影响对行为和对小鼠适应实验环境，包含小鼠的笼从该动物保持室转移到相邻的一个隔音测试室，至少30分钟的隔音候车室每次测试开始之前。 </li><li>所有的实验（ 图2A）的应在s中被执行每天火焰的时间段中的光或暗相位以减少由在不同时间23,24测试所产生的行为变化。在这个协议中，所有的实验都是轻相和1小时发病的黑暗阶段（上午8:00至下午6:00在轻相）发病前1小时后之间进行。如果只有一个设备是可用的，每个基因型的小鼠，应在一个平衡，以减少试验时间和对行为表现受试者的测试顺序的潜在影响进行试验。 ImageFZ可以控制最多4个装置。同时测试4只小鼠用4设备的平衡才能使研究人员节省时间，降低了实验参数对小鼠行为的可能影响。 </li></ol> 3。空调<ol><li>小鼠被放置在调节室和小鼠通常允许自由地探索该腔室为120秒。此后，非盟ditory球杆，例如白噪声，音调和听觉发声，是作为一个CS，持续30秒，最后2秒的声音的过程中0.1-0.8毫安电击被给予小鼠作为美国。在CS-US配对的呈现反复强化的关联。将小鼠留在腔室中的时间长度，最后介绍后，进一步建立该室的上下文和厌恶的经验之间的关联。在这个协议中，120秒的自由探索后，听觉提示（白噪声55 dB）被提出，持续30秒，并且一个0.3毫安电击的最后2秒的白噪声的过程中连续地传递。 90秒后，将听觉线索与电击的配对被再次提供给受试者。 CS-US的呈现重复每届3次（120，240，和360秒的调理开始后）（ 图2B）。以下最后的电击，将小鼠在腔室90秒静置。 </li><li>调理会话开始之前，运行ImageFZ应用软件程序，选择插件菜单“FZ空调和FZ在线（4室）”，并设置参数值一步一步如下。 <ol><li>第1步：项目编号。指定要在其中存储数据文​​件的文件夹。 </li><li>步骤2：会话名称。输入任何文字， 如实验数据，在“会话”对话框，然后选择其中的起始时间和白噪声和电击的时间都写，在“参考”框中键入一个参考的文本文件。样本的文本文件显示在视频。 </li><li>第3步：参数设置。在每个方块中输入如下的参数值。 <ol><li>速度（帧/秒）：图像采集的帧率， 比如 1帧/秒。 </li><li>持续时间（秒）：在空调的情况下，总的持续时间为480秒。 </li><li>选区持续时间（秒）： 例如 60秒;的数据在每个块中进行分析的60秒。 </li><li>主题大小 - 最小（像素）：ImageFZ检测到鼠标和噪声黑色颗粒（像素部分质量）在一个白色的背景在每个图像。当黑色颗粒（像素）的面积是小于“主题大小-分钟（像素数）”的值（ 例如 100像素）时，颗粒被视为噪声而被排除在图像分析。 </li><li>被摄体尺寸 - 最大（像素）：当黑色颗粒的尺寸超过了“主题大小 - 最大（像素）&#39;的尺寸值时，颗粒被从分析中排除。 </li><li>帧大小-宽度/高度（厘米）：室尺寸， 即 33厘米宽和25厘米高。 </li><li>冻结准则（像素）： 例如 30像素，看到协议6的细节。 </li><li>冻结时间-分钟（秒）： 如 2秒，当检测到只有不到2秒无鼠标移动，其行为不算作“冻结”;。 </li><li>冲击速度（帧/秒）：见协议6的细节。 </li></ol></li><li>第4步：主题标识。输入对象的识别。 </li><li>步骤5：相机设置。控制亮度和所捕获的图像的对比度。 </li><li>第6步：阈值设置。调整阈值来检测黑色的鼠标为黑色像素在白色背景中的每个图像，并判断鼠标行为为“冻结”或“非冻结”（见议定书6的细节）。分析白化鼠标点击复选框“反转模式”，并适当调整阈值。 </li><li>第7步：设置网箱球场。指定你想要捕捉的每个室的领域。单击工具箱中的矩形按钮后，周围绘制实时图像窗口中室的地板的矩形。接下来，选择室号，然后单击“设置”按钮。最后，点击“完成”按钮。 </li></ol></li> &lt;LI&gt;后的参数设置选项，预备试验应采用的做法小鼠（不作为主题Mice）当天的先行先试，以确定图像分析系统和白噪音/震动发电机是否没有问题的工作给予。 </li><li>移动包含实践小鼠对从相邻的候车室的隔音测试室一个家笼，然后将每只小鼠的调节室。立即把小鼠的室之后，单击ImageFZ的启动按钮。该应用软件将在您的参考文件中指定的顺序呈现听觉线索和/或电footshocks给小鼠。 </li><li>后480秒已经过去时，返回的小鼠其家笼和笼返回到在保持室的架子上。 </li><li>仔细清洁室。然后，单击“下一步分析”按钮，并重复步骤3.2.4-3.6对试验鼠。 </li><li> ImageFZ店住和TIFF格式的图像跟踪。该计划允许我们执行脱机分析使用修改后的参数值来重新分析图像。如果你进行脱机分析，选择插件菜单中的“恐惧条件和FZ离线”，然后选择您要重新分析数据文件夹。此后，输入的参数值再次，然后点击“完成”按钮。 </li></ol> 4。语境测试<ol><li>调理会话完成后，将小鼠返回到相同的调理室，并取得冻结行为来衡量语境条件恐惧（上下文测试）。调理和上下文测试之间的延迟间隔已被普遍设定为24小时。在这个协议中，以评估近期记忆和远期记忆25（由测试1天，调理后28天，分别测得），小鼠受到上下文测试约24小时，30天调理会议之后。将小鼠放置在已调节itioning室，并允许自由探索室为300秒，而没有CS和美国演讲（ 图2C）。 </li><li>运行ImageFZ软件程序和以同样的方式设置了应用软件的参数值，如在调节（见3.2.3节），但是，修改该试验的持续时间为300秒，并选择上下文测试的参考文本文件。更改设置后，预备试验应采用的做法老鼠来检查ImageFZ系统给出。 </li><li>将每只小鼠放入调理室，然后点击开始按钮。经过300秒过去后，返回小鼠到他们家笼，并留下笼原状，直到线索测试开始。 </li><li>清洁室。然后，单击“下一步分析”按钮，并重复步骤4.3-4.4在试验小鼠。 </li></ol> 5。线索测试<ol><li>线索测试上下文测试的当天或第二天进行。在该试验中，小鼠被放置到另一个测试腔室具有非常不同的特性，提供了一种新的上下文无关的调节室3分钟。在第一个3分钟结束时，即，提出了在空调时的听觉线索是给老鼠在小说的上下文环境中3分钟。在这个协议中，线索执行测试的上下文中试验后的几个小时。老鼠被允许探索三角形室360秒。在第3分钟，既不是CS，或是美国提出，此后，一个CS（一55分贝白噪声），提出在最后3分钟。 </li><li>运行ImageFZ软件程序和设置的参数值中的相同方式在空调，除了修改测试的持续时间，以360秒，然后为线索测试的参考文本文件。调整设置后，预备试验应采用的做法老鼠来检查ImageFZ系统给出。 </li><li>将每只小鼠放入三角腔，然后点击开始按钮。后360秒已经过去时，返回的小鼠其家笼和笼返回到收容室的架子上。 </li><li>清洁室。然后，单击“下一步分析”按钮，并重复步骤5.3-5.4在试验小鼠。 </li><li>为了进一步测试远程内存，空调会议（ 图2A）4-5后30天左右重复的协议。 </li></ol> 6。图像分析<ol><li>自动使用ImageFZ进行数据采集和分析。该应用软件程序是基于公共领域的ImageJ程序（由韦恩Rasband在卫生部和国家机构可在http://rsb.info.nih.gov/ij/开发），由宫川刚修改（ImageFZ应用软件，可以免费下载，见表材料/试剂）。 </li><li>对于所有实验，采集图像在使用USB视频捕获设备，包括一个给定的帧速率（ 例如 1帧）与ImageFZ视频摄像头。为了测量从连续的图像移动的距离，调整“门槛最小”值的程序（ 例如 80像素），它被设置为段影像变成了黑色颗粒（鼠标）和一个白色的背景。行进的距离从各组在xy间的距离的坐标为粒子的重心在连续图像的中心来计算。 </li><li>为了测量从连续图像的冻结行为，调整“门槛分钟（XOR）价值的程序（ 例如 160像素），它被设置为段影像变成了黑色颗粒（鼠标）和背景，然后计算金额每对连续图像的颗粒之间不重叠区域的面积（像素）。使用阈值工具的滑块，直到黑色粒子在每个图像中的鼠标不包括​​尾部的整个身体的形状相匹配调整值。如果nonoverla的面积pping区域下方的“冷冻准则”的值（ 例如 30个像素），则该行为被认为是“冻结”（ 图3），其通常被定义为除了呼吸和心跳完全没有任何运动。当面积超过该值，则该行为被认为是“非冷冻&#39;（ 图3）。判决应当根据冷冻的定义进行。老鼠有时表现出细微的动作和瞬间不动，这可能不被视为一个冻结行为，反映了恐惧。固定性，持续的时间很短（ 例如小于2秒），这是从恐惧的表现可能会有所不同，可以通过设置冷冻的时间阈值被排除在分析之外。要设置时间阈值，输入&#39;冻结时间-分钟（秒）&#39;的值（ 例如 2秒）。 </li><li>该ImageFZ程序自动演算阿泰行驶的距离（厘米）和冷冻的百分比。结果保存在文本文件中，生活和跟踪图像存储在一个TIFF格式。为了测量行驶距离（厘米）电动电击敏感的指标，该ImageFZ方案还处于较高的帧速率（ 例如 4帧），持续6秒，2秒测量的2秒电击的交付前，直到2获取的图像秒内在线分析电击后。之前设置的帧频图像采集，期间和之后，电击，在输入一个值“冲击速度（帧/秒）”复选框。在线分析后，通过选择插件菜单中的“FZ冲击离线”，以获得行驶的距离数据进行离线分析。 </li><li>该ImageFZ程序的参数值应进行优化，以产生类似的结果由人类观察者在预备测试中获得。对于手动得分，冻结行为不断使用stopwatc测H和事件记录程序或瞬时时间抽样程序每3-10秒，在此期间使用ImageFZ软件分析。两个观察者通常进行行为观察。要调整ImageFZ程序的参数值，以确保图像分析的结果是与人类观察者一致，执行ImageFZ程序的离线分析，修改了“门槛分钟（XOR）”和“冻结标准的价值观。为了进行离线分析，选择插件菜单中的“FZ离线&#39;和输入任何参数值。 </li></ol> 7。故障排除<ol><li>怎样才能ImageFZ程序获得并安装？ 该ImageFZ程序是可以从我们的网站免费下载（见表材料/试剂），并运行在Windows计算机上。下载zip文件夹ImageFZ和您的计算机上安装该软件。见&#39;的readme.txt&#39;文件的installati有关详细信息，并按照一步一步的指示。 </li><li>为什么显示错误消息“错误的设置捕获设备”？ 检查摄像机电缆的连接和驱动程序安装的USB图像采集设备的。如果没有与该设置没有问题，那么ImageFZ软件可能无法与您的图像捕获设备正常工作。见关于相应的设备与ImageFZ软件使用&#39;的readme.txt“文件。 </li><li> ImageFZ不能检测到鼠标的整个身体的颗粒。 设置的“门槛分钟”和/或“门槛分钟（XOR）”比当前值小的值。如果ImageFZ不能检测到鼠标在一个特定的地方， 例如 ，一个测试室的角落里，然后测试条件不足，如一个均匀地照明的地板或鼠标和背景之间的反差稍微的差异，可能存在。为了解决这个问题，调整参数的值（ 例如 </eImageFZ的米&gt;亮度和实时图像和阈值的对比），控制灯的数量和位置，或者使用白色背景的黑色的鼠标。 </li><li>以高帧率图像拍摄过程中的在线分析会减慢计算机。 帧速率设定为比目前的速度低，并且执行在线分析。该ImageFZ分析，通过在1帧​​的图像采集，足以准确地测量冷冻，如图代表性结果部分。 </li><li>该ImageFZ分析的结果不同意那些人的得分。 检查存储的图像和判断结果文件。如果ImageFZ高估冻结的&#39;冻结准则“的值设置为大于当前值的情况下，与进行离线分析。如果ImageFZ低估冷冻的&#39;冻结准则“的值设置为比当前值更高。 </li><li>在optogenetical和体内电éxperiments，附着在小鼠头部的光纤电缆干扰冷冻的判断。 外套的电缆在白色的黑色的鼠标，改变相机的位置和角度，直到未检测到电缆。 </li><li>什么是需要的离线分析？ 创建一个名为“Image_FZ”在ImageFZ程序的根目录文件夹。在这个文件夹中创建子文件夹“图片”和“会话”。移动8位灰度图像到“图片”文件夹，并创建了图像的文件名是写在“会话”文件夹中的文本文件。此后，运行ImageFZ离线分析，并按照程序的指示。 </li></ol>

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	<li>Crawley, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> What's wrong with my mouse: behavioral phenotyping of transgenic and knockout mice. , (2007).</li><li>Fanselow, M. S., Poulos, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neuroscience+of+mammalian+associative+learning.">The neuroscience of mammalian associative learning.</a> Annu. Rev. Psychol. 56, 207-234 (2005).</li><li>LeDoux, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emotion+circuits+in+the+brain.">Emotion circuits in the brain.</a> Annu. Rev. Neurosci. 23, 155-184 (2000).</li><li>Nakajima, R., 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=Comprehensive+behavioral+phenotyping+of+calpastatin-knockout+mice.">Comprehensive behavioral phenotyping of calpastatin-knockout mice.</a> Mol. Brain. 1, 7 (2008).</li><li>Ikeda, M., 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=Identification+of+YWHAE,+a+gene+encoding+14-3-3epsilon,+as+a+possible+susceptibility+gene+for+schizophrenia.">Identification of YWHAE, a gene encoding 14-3-3epsilon, as a possible susceptibility gene for schizophrenia.</a> Hum. Mol. Genet. 17, 3212-3222 (2008).</li><li>Sakae, N., 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=Mice+lacking+the+schizophrenia-associated+protein+FEZ1+manifest+hyperactivity+and+enhanced+responsiveness+to+psychostimulants.">Mice lacking the schizophrenia-associated protein FEZ1 manifest hyperactivity and enhanced responsiveness to psychostimulants.</a> Hum. Mol. Genet. 17, 3191-3203 (2008).</li><li>Fukuda, 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=Down-regulation+of+protocadherin-alpha+A+isoforms+in+mice+changes+contextual+fear+conditioning+and+spatial+working+memory.">Down-regulation of protocadherin-alpha A isoforms in mice changes contextual fear conditioning and spatial working memory.</a> Eur. J. Neurosci. 28, 1362-1376 (2008).</li><li>Imayoshi, I., 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=Roles+of+continuous+neurogenesis+in+the+structural+and+functional+integrity+of+the+adult+forebrain.">Roles of continuous neurogenesis in the structural and functional integrity of the adult forebrain.</a> Nat. Neurosci. 11, 1153-1161 (2008).</li><li>Nakatani, 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=Abnormal+behavior+in+a+chromosome-engineered+mouse+model+for+human+15q11-13+duplication+seen+in+autism.">Abnormal behavior in a chromosome-engineered mouse model for human 15q11-13 duplication seen in autism.</a> Cell. 137, 1235-1246 (2009).</li><li>Takao, K., 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=Comprehensive+behavioral+analysis+of+calcium/calmodulin-dependent+protein+kinase+IV+Knockout+mice.">Comprehensive behavioral analysis of calcium/calmodulin-dependent protein kinase IV Knockout mice.</a> PLoS ONE. 5, (2010).</li><li>Tamada, K., 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=Decreased+exploratory+activity+in+a+mouse+model+of+15q+duplication+syndrome;+implications+for+disturbance+of+serotonin+signaling.">Decreased exploratory activity in a mouse model of 15q duplication syndrome; implications for disturbance of serotonin signaling.</a> PLoS ONE. 5, (2010).</li><li>Watanabe, Y., 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=Relaxin-3-deficient+mice+showed+slight+alteration+in+anxiety-related+behavior.">Relaxin-3-deficient mice showed slight alteration in anxiety-related behavior.</a> Front. Behav. Neurosci. 5, 50 (2011).</li><li>Takeuchi, 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=P301S+mutant+human+tau+transgenic+mice+manifest+early+symptoms+of+human+tauopathies+with+dementia+and+altered+sensorimotor+gating.">P301S mutant human tau transgenic mice manifest early symptoms of human tauopathies with dementia and altered sensorimotor gating.</a> PLoS ONE. 6, (2011).</li><li>Koshimizu, 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=Adenomatous+polyposis+coli+heterozygous+knockout+mice+display+hypoactivity+and+age-dependent+working+memory+deficits.">Adenomatous polyposis coli heterozygous knockout mice display hypoactivity and age-dependent working memory deficits.</a> Front. Behav. Neurosci. 5, 85 (2011).</li><li>Yao, I., Takao, K., Miyakawa, T., Ito, S., Setou, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synaptic+E3+ligase+SCRAPPER+in+contextual+fear+conditioning:+extensive+behavioral+phenotyping+of+Scrapper+heterozygote+and+overexpressing+mutant+mice.">Synaptic E3 ligase SCRAPPER in contextual fear conditioning: extensive behavioral phenotyping of Scrapper heterozygote and overexpressing mutant mice.</a> PLoS ONE. 6, (2011).</li><li>Shoji, 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=Comprehensive+behavioral+analysis+of+ENU-induced+Disc1-Q31L+and+-L100P+mutant+mice.">Comprehensive behavioral analysis of ENU-induced Disc1-Q31L and -L100P mutant mice.</a> BMC Res. Notes. 5, 108 (2012).</li><li>Anagnostaras, S. G., Josselyn, S. A., Frankland, P. W., Silva, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Computer-assisted+behavioral+assessment+of+Pavlovian+fear+conditioning+in+mice.">Computer-assisted behavioral assessment of Pavlovian fear conditioning in mice.</a> Learn. Mem. 7, 58-72 (2000).</li><li>Miyakawa, T., Yamada, M., Duttaroy, A., Wess, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperactivity+and+intact+hippocampus-dependent+learning+in+mice+lacking+the+M1+muscarinic+acetylcholine+receptor.">Hyperactivity and intact hippocampus-dependent learning in mice lacking the M1 muscarinic acetylcholine receptor.</a> J. Neurosci. 21, 5239-5250 (2001).</li><li>Marchand, A. R., Luck, D., DiScala, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+an+improved+automated+analysis+of+freezing+behaviour+in+rats+and+its+use+in+trace+fear+conditioning.">Evaluation of an improved automated analysis of freezing behaviour in rats and its use in trace fear conditioning.</a> J. Neurosci. Methods. 126, 145-153 (2003).</li><li>Kopec, C. D., 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=A+robust+automated+method+to+analyze+rodent+motion+during+fear+conditioning.">A robust automated method to analyze rodent motion during fear conditioning.</a> Neuropharmacology. 52, 228-233 (2007).</li><li>Wehner, J. M., 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=Quantitative+trait+locus+analysis+of+contextual+fear+conditioning+in+mice.">Quantitative trait locus analysis of contextual fear conditioning in mice.</a> Nat. Genet. 17, 331-334 (1997).</li><li>Quirk, G. J., Armony, J. L., LeDoux, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fear+conditioning+enhances+different+temporal+components+of+tone-evoked+spike+trains+in+auditory+cortex+and+lateral+amygdala.">Fear conditioning enhances different temporal components of tone-evoked spike trains in auditory cortex and lateral amygdala.</a> Neuron. 19, 613-624 (1997).</li><li>Chaudhury, D., Christopher, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Circadian+modulation+of+learning+and+memory+in+fear-conditioned+mice.">Circadian modulation of learning and memory in fear-conditioned mice.</a> Behav. Brain Res. 133, 95-108 (2002).</li><li>Valentinuzzi, V. S., 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=Effect+of+circadian+phase+on+context+and+cued+fear+conditioning+in+C57BL/6J+mice.">Effect of circadian phase on context and cued fear conditioning in C57BL/6J mice.</a> Learn. Behav. 29, 133-142 (2001).</li><li>Frankland, P. W., Bontempi, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+organization+of+recent+and+remote+memories.">The organization of recent and remote memories.</a> Nat. Rev. Neurosci. 6, 119-130 (2005).</li><li>Contarino, A., Baca, L., Kennelly, A., Gold, L. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+assessment+of+conditioning+parameters+for+context+and+cued+fear+in+mice.">Automated assessment of conditioning parameters for context and cued fear in mice.</a> Learn. Mem. 9, 89-96 (2002).</li><li>Kinney, J. W., 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=Deficits+in+trace+cued+fear+conditioning+in+galanin-treated+rats+and+galanin-overexpressing+transgenic+mice.">Deficits in trace cued fear conditioning in galanin-treated rats and galanin-overexpressing transgenic mice.</a> Learn. Mem. 9, 178-190 (2002).</li><li>Hefner, K., Holmes, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ontogeny+of+fear-,+anxiety-+and+depression-related+behavior+across+adolescence+in+C57BL/6J+mice.">Ontogeny of fear-, anxiety- and depression-related behavior across adolescence in C57BL/6J mice.</a> Behav. Brain Res. 176, 210-215 (2007).</li><li>Wellman, C. L., 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=Impaired+stress-coping+and+fear+extinction+and+abnormal+corticolimbic+morphology+in+serotonin+transporter+knock-out+mice.">Impaired stress-coping and fear extinction and abnormal corticolimbic morphology in serotonin transporter knock-out mice.</a> J. Neurosci. 27, 684-691 (2007).</li><li>Valentinuzzi, V. S., 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=Automated+measurement+of+mouse+freezing+behavior+and+its+use+for+quantitative+trait+locus+analysis+of+contextual+fear+conditioning+in+(BALB/cJ+&#215;+C57BL/6J)F2+mice.">Automated measurement of mouse freezing behavior and its use for quantitative trait locus analysis of contextual fear conditioning in (BALB/cJ &#215; C57BL/6J)F2 mice.</a> Learn. Mem. 5, 391-403 (1998).</li><li>Valentinuzzi, V. S., 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=Effect+of+circadian+phase+on+context+and+cued+fear+conditioning+in+C57BL/6J+mice.">Effect of circadian phase on context and cued fear conditioning in C57BL/6J mice.</a> Learn. Behav. 29, 133-142 (2001).</li><li>Bothe, G. W. M., Bolivar, V. J., Vedder, M. J., Geistfeld, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+and+behavioral+differences+among+five+inbred+mouse+strains+commonly+used+in+the+production+of+transgenic+and+knockout+mice.">Genetic and behavioral differences among five inbred mouse strains commonly used in the production of transgenic and knockout mice.</a> Genes Brain Behav. 3, 149-157 (2004).</li><li>Chen, C., Kim, J. J., Thompson, R. F., Tonegawa, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hippocampal+lesions+impair+contextual+fear+conditioning+in+two+strains+of+mice.">Hippocampal lesions impair contextual fear conditioning in two strains of mice.</a> Behav. Neurosci. 110, 1177-1180 (1996).</li><li>Anagnostaras, S. G., Gale, G. D., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hippocampus+and+contextual+fear+conditioning:+Recent+controversies+and+advances.">Hippocampus and contextual fear conditioning: Recent controversies and advances.</a> Hippocampus. 11, 8-17 (2001).</li><li>Akirav, I., Maroun, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+the+medial+prefrontal+cortex-amygdala+circuit+in+stress+effects+on+the+extinction+of+fear.">The role of the medial prefrontal cortex-amygdala circuit in stress effects on the extinction of fear.</a> Neural Plast. , 1-11 (2007).</li><li>Johansen, J. P., Cain, C. K., Ostroff, L. E., LeDoux, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+mechanisms+of+fear+learning+and+memory.">Molecular mechanisms of fear learning and memory.</a> Cell. 147, 509-524 (2011).</li><li>Anagnostaras, S. G., 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=Automated+assessment+of+pavlovian+conditioned+freezing+and+shock+reactivity+in+mice+using+the+video+freeze+system.">Automated assessment of pavlovian conditioned freezing and shock reactivity in mice using the video freeze system.</a> Front. Behav. Neurosci. 4, (2010).</li><li>Pham, J., Cabrera, S. M., Sanchis-Segura, C., Wood, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+scoring+of+fear-related+behavior+using+EthoVision+software.">Automated scoring of fear-related behavior using EthoVision software.</a> J. Neurosci. Methods. 178, 323-326 (2009).</li><li>Blanchard, D. C., Blanchard, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Crouching+as+an+index+of+fear.">Crouching as an index of fear.</a> J. Comp. Physiol. Psychol. 67, 370-375 (1969).</li><li>Frankland, P. W., Bontempi, B., Talton, L. E., Kaczmarek, L., Silva, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+involvement+of+the+anterior+cingulate+cortex+in+remote+contextual+fear+memory.">The involvement of the anterior cingulate cortex in remote contextual fear memory.</a> Science. 304, 881-883 (2004).</li><li>Takao, K., Miyakawa, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigating+gene-to-behavior+pathways+in+psychiatric+disorders:+the+use+of+a+comprehensive+behavioral+test+battery+on+genetically+engineered+mice.">Investigating gene-to-behavior pathways in psychiatric disorders: the use of a comprehensive behavioral test battery on genetically engineered mice.</a> Ann. N. Y. Acad. Sci. 1086, 144-159 (2006).</li><li>Takao, K., Yamasaki, N., Miyakawa, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+brain-behavior+phenotypying+of+genetically-engineered+mice+on+research+of+neuropsychiatric+disorders.">Impact of brain-behavior phenotypying of genetically-engineered mice on research of neuropsychiatric disorders.</a> Neurosci. Res. 58, 124-132 (2007).</li></ol>

本公司确认，目前并无任何已知的利益冲突，与此相关的出版物，并出现了对这项工作有任何足以影响其结果并无显著的财政支持。

上下文和线索恐惧条件测试是使用最广泛的范式来评估学习和记忆之一。这个测试是巴甫洛夫条件反射的一种形式，其中一个关联的上下文和/或条件刺激（听觉提示）和厌恶刺激（电电击）之间进行。上下文/听觉线索和电击，甚至一个单一的配对后，小鼠表现出持久的冰点时，面对无论是内容或提示。在该试验中，冷冻的行为被用作恐惧记忆的指数。药理和病变的研究发现，记忆的形成，巩固和检索由几个大脑区域，如杏仁核，海马和前额叶皮层3,33-35监管。此外，分子遗传学研究已经证明特定的基因以及参与在用遗传工程小鼠36这些脑区的学习和记忆的分子的作用。因此，这种测试是简单程序e和探索的神经生物学基础底层的恐惧学习和记忆非常有用。在这部影片中文章中，我们介绍了我们的协议来提供实验者提供详细的信息，以理解和容易地进行测试。 冻结行为是通过直接观察人类实验者量化。一个训练有素的实验者预计将产生跨越观测可靠，稳定的结果。然而，这种方法涉及到潜在的问题，例如在观测方法的差异，观察者偏倚和简单量化的错误，从而难以直接结果从独立实验者和不同的实验室比较。一个自动化的光子束为基础的计算机测量系统也被用来26,30-32。然而，这种系统也存在于测定冻结行为的潜在问题。由于传感器装置，该系统可能无法检测小头部动作，将typicaLLY评价为“主动”通过人的观察。此外，凝固过程中颤抖着可能被视为非冻结，因为当动物冻结，在光子束间歇性中断被观察到颤抖的后果。作为一种替代方法，自动图像和视频分析系统已经开发17-20,37,38。 Anagnostaras 等 37所描述的几个系统具有良好的有效性和成绩冻结以及17,20,37-38图像分析软件程序。然而，大多数这些系统和分​​析程序必须从商业供应商获得，并且通常昂贵。我们开发的ImageFZ软件程序的冻结行为的分析，并将该软件分发作为一个免费的软件程序。 ImageFZ检测小鼠为像素的主体（颗粒），并判别细微鼠标移动，“冻结”或“非冻结”视额每对连续图像的颗粒之间不重叠的区域的面积。如图所示，在具有代表性的结果，使用ImageFZ程序测量与一致或比用其它方法得到的更加精确。因此，ImageFZ程序自动测量的行为，人类观察者判断，使用定义的标准冻结。此外，ImageFZ程序计算行驶距离（厘米）之前，期间和电击暴露后，促进冻结行为的冲击灵敏度和分析进行评估。 方法上的差异实验室间存在的。这些差异可能会导致难以实验室之间的数据进行比较，并在不同的实验室复制的结果。以获得更稳定的和可比较的数据，有必要标准化的测试协议尽可能。该分析系统具有ImageFZ导致的测试程序的自动化，从而可以向协议的标准化用于整个实验室。 在分析冻结行为的若干行为反应必须加以考虑。首先，当动物面临一个可怕的情况，他们可能逃跑，而不是冻结39。逃跑是恐惧反应之一，它的发生会导致低估的恐惧记忆。第二，冻结可能依赖于一般的活性水平，并在实验和对照组小鼠的活动级别有待检验。例如，尽管缺乏M1毒蕈碱型乙酰胆碱受体的小鼠表现出对低温较野生型小鼠的水平降低，各种行为测试表明，结果可能是由于他们的多动型的，而不是他们的记忆障碍18。 ImageFZ计算由主体行进的距离（厘米）。这些数据可用来检查对象之间的一般活动量是否不存在差异。如果有一组差行驶的距离，一个可能的解决问题的方法是考虑训练的前2分钟为基线活动期间的行驶距离，并在基线+活动期间使用抑制比（在测试过程中抑制率=（活性）/（活动测试））的恐惧17,40二级索引。最后，在疼痛敏感性的差异，诱导一种电动电击的变化，反应性，如果有的话，可能会导致变化的凝固行为的影响。 ImageFZ还计算的距离从2秒行进（厘米）的详细情况的2秒电击的曝光到2秒的曝光（6秒）后，它可以被用来作为电击灵敏度的指数前。 视频分析系统已被开发来测量白化，黑色，刺豚鼠的凝固行为，并稀释棕色小鼠。 ImageFZ采用黑色地板托盘和黑色网格来检查白小鼠（参见图1B）。黑色网格由SPECI的<html>同盟加工金属带涂层的黑色涂料和具有类似于未涂覆的金属网格，这通常用于黑色小鼠的电导率。 ImageFZ还分析了老鼠等啮齿类动物通过程序参数的调整，冻结行为。在ImageFZ的当前版本中，受试者的行为用摄像机从顶壁来分析冷冻被记录。 ImageFZ也可以在一个设置，其中该图像从所述腔室的一侧拍摄的使用。此外，ImageFZ控制最多4装置。此功能允许研究人员同时检测4只小鼠，节省了时间和从每个受试者的执行时间和对行为的测试顺序的差异减少了潜在的影响。因此，ImageFZ简化了测试程序，并冻结行为分析，并将该软件方便测试用更少的劳动力和没有对行为实验的任何培训。 e_content“&gt;在宫川的实验室，我们已经评估了超过110株使用视频分析系统阐明某一基因对学习记忆的影响遗传工程小鼠和野生型对照组小鼠的背景和线索恐惧条件测试41-42我们已经获得一大套的原始数据为5000多老鼠被用于发表的研究文章4-16的原始数据都包含在&#39;鼠表型数据库“作为一个公共数据库（网址：。的http： / / www.mouse-phenotype.org/）。本电影的文章提供了有关我们的实验过程的细节的详细信息和促进的测试情况的​​了解。

上下文和线索恐惧条件测试是用来评估在啮齿类动物中1-3关联恐惧学习和记忆的行为模式。该测试已被广泛用于了解转基因和基因敲除小鼠1,4-16恐惧学习和记忆的神经生物学机制。冻结行为，其被定义为完全不动，除了呼吸，是可怕的情况的共同反应。在这种行为范式，后动物暴露于听觉提示用电动电击的配对，他们的恐惧产生的刺激通过显示冻结行为，这是衡量联想恐惧学习和记忆的索引回应。这个测试需要不太复杂的设备，由研究者少体力消耗，以及对小鼠比其他的学习和记忆任务要少得多的训练时间，所以一般2天大约需要5-10元鼠标分钟/天。虽然测试程序是SI投影机的简易并且需要很少的时间来执行，调查员必须仔细观察和测量鼠标行为，因此，一些自动化测量系统已经发展到进行行为分析17-20。我们的视频分析系统，我们开发了与ImageFZ软件程序，使我们能够轻松地分析冻结行为，并产生高度可靠的结果。本文提供我们的测试过程的详细信息，并介绍如何使用ImageFZ软件程序。

<table border="0" cellpadding="0" cellspacing="0" width="701">
	<tbody>
		<tr>
			<td>&#160;ImageFZ&#160; program</td>
			<td>Developed by Tsuyoshi Miyakawa</td>
			<td></td>
			<td>&#160;This program is available through O&#39;Hara &#38; Co., Tokyo, Japan and for free download at http://www.mouse-phenotype.org/. This software runs on 32-bit Windows XP/Vista/7.&#160;</td>
		</tr>
		<tr>
			<td>Conditioning chamber</td>
			<td>O&#39;Hara &#38; co., Japan</td>
			<td>CL-3002L</td>
			<td>For mouse.</td>
		</tr>
		<tr>
			<td>Cued test chamber</td>
			<td>O&#39;Hara &#38; co., Japan</td>
			<td>CLT-3002L</td>
			<td>For mouse.</td>
		</tr>
		<tr>
			<td>Interface</td>
			<td>O&#39;Hara &#38; co., Japan</td>
			<td>CL-1040</td>
			<td>The interface includes a white noise/tone generator, which can be controlled by ImageFZ program.</td>
		</tr>
		<tr>
			<td>Scrambled shock generator</td>
			<td>O&#39;Hara &#38; co., Japan</td>
			<td>SGA-2040</td>
			<td>The shock generator can be controlled by ImageFZ program.</td>
		</tr>
		<tr>
			<td>Shock grid tester (ammeter)</td>
			<td>O&#39;Hara &#38; co., Japan</td>
			<td>SG-T</td>
			<td></td>
		</tr>
		<tr>
			<td>USB video capture device</td>
			<td>XLR8</td>
			<td>USB2IVOSX</td>
			<td></td>
		</tr>
		<tr>
			<td>Quad image splitter</td>
			<td>Wireless Tsukamoto Co., Ltd., Japan</td>
			<td>400AS</td>
			<td></td>
		</tr>
		<tr>
			<td>Soundproof room</td>
			<td>O&#39;Hara &#38; co., Japan</td>
			<td>CL-4210</td>
			<td></td>
		</tr>
		<tr>
			<td>Freeze Monitor</td>
			<td>San Diego Instruments, Inc., CA, USA</td>
			<td></td>
			<td>16 x 16 photbeam array&#160; ( 2.5 cm spacing)</td>
		</tr>
	</tbody>
</table>

contextual and cued fear conditioning test using a video analyzing system in mice

上下文和线索恐惧条件测试是行为测试，评估小鼠的学习和记忆的环境暗示和厌恶体验之间的关联的能力之一。在该试验中，将小鼠置于一个温度调节室，并给予条件刺激（听觉提示）的碎料和厌恶的非条件刺激（电电击）。经过一段延迟时间，将小鼠暴露于相同的调节室和一个不同形状的腔室与上介绍的听觉线索的。在测试过程中冻结行为被测量为恐惧记忆的指数。自动分析的行为，我们已经开发使用ImageFZ应用软件程序，它可于http://www.mouse-phenotype.org/免费下载的视频分析系统。在这里，为了表示我们的协议的细节，我们展示了我们的程序的上下文，并使用ImageFZ系统·线索恐惧条件测试在C57BL/6J小鼠EM。此外，我们证实了我们的协议和通过比较由ImageFZ系统或一个光子束的计算机测量系统与该计分由人类观察者测定冷冻时间的视频分析系统的性能。如在我们的代表性结果，由ImageFZ获得的数据类似于那些由人类观察者分析，表明使用ImageFZ系统的行为分析是高度可靠的。本影片本文提供有关测试过程的详细信息，并会促进实验了解情况。

在恐惧条件测试中，人类实验者使用通过劳动密集的直接观察26-29量化凝固行为的影响，但最近光子束为基础的计算机的测量（ 如 &#39;冻结显示器“系统）和图像分析系统已被用于自动测量冻结行为26,30-32。 ImageFZ是一种自动化图像分析系统，其产生的结果媲美通过人的观察得到的，如下文所述。在这里，我们比较了人类观察在不同参数的ImageFZ分析的结果：“速度（帧/秒）&#39;和&#39;冻结准则（像素）。”在这个实验中，雄性C57BL/6J小鼠（平均体重±标准差（克），31.4±3.55，平均机身尺寸为±标准差（像素），351.6±62.2）被用来在15-27周龄。人类观测在使用事件记录程序（在Macintosh OS9软件程序）制成，即持续一个键按压事件d表示2秒以上时，鼠标显示的没有运动的一个回合被认为是“冻结”。冷冻的百分比计算，每60秒在每个测试和用于相关性分析。进球结冰的2名观察员的百分比（观察者间的可靠性，进行调理，R = 0.879;的情况下测试，R = 0.957;为线索测试，R = 0.866，对于所有的情况下，R = 0.888）的平均值来产生人类得分。通过ImageFZ与每个帧速率（ 即 1，2，和4 fps）的测得的冷冻百分数和那些通过人的观察所得之间的相关性进行了研究。如如图4所示，通过ImageFZ（1，2，和4 fps）的计算出的冷冻率的结果高度相关，与从2观察者的测量获得的平均值。值得注意的是，在拍摄图像以较高的帧速率并不总能产生最好的相关性。在1帧产生类似的结果从人类观察者在电子商务获得的图像分析阿赫测试。通过人的观察测定，并使用ImageFZ的各条件下“冷冻条件（像素）&#39;冻结百分比（ 即 20，30，和40个像素）之间的相关性进行了研究。使用ImageFZ在20&#39;冷冻条件（像素）&#39;，30，和40个像素计算出的冷冻率的结果，在所有的情况下，高度相关的那些通过人的观察（ 图5）获得。 如图5D所示 ，当冷冻准则设置为一个低的值，鼠标的微妙运动，认为是“冻结”由人类观察者，将被认为是使用ImageFZ&#39;非冻结“。相反，如果标准被设定为较高的值，鼠标的移动，评定为“非冻结”人类观察员，将被视为使用ImageFZ（ 图5C，5F，以及5I）“冻结”。因此，为了获得最可靠的结果，本ImageFZ程序的各参数应校准我们ING通过人的观察在每个测试环境中打进的数据。 此外，我们比较了由人类观察者所产生的效果，使用光子束的计算机测量系统（冻结监控系统）中，与使用ImageFZ得到的（ 见图6）。人类观察者被蒙蔽治疗组和ImageFZ打分的结果。对于冻结监视器系统的参数设置，我们使用了从以前验证系统30冻结的百分比3措施。简言之，将10秒的时间间隔的数目，其中动物需要多于1或2秒到相互交叉5秒时间间隔的开始之间的时间间隔（1秒10秒和2秒10秒，分别）和延迟的第一个新的光束和这个区间（Latency3）内的第三个新的光束中断测量。在此期间，将小鼠冻结间隔或百分比时间r的总量的百分比必修打破第三光子束进行了计算。 冷冻的每个系统中测得的比例示于图6。使用两种方式重复测量ANOVA，然后通过t-检验（ 见表1）进行比较的基团。使用ImageFZ（ 图6B）测定的冷冻百分比分别为更类似于那些计分，通过人的观察（ 图6A）比使用光子束为基础的系统（ 图6C-E）获得的数据。 ，而冻结的百分比之间的相关性测量每个测试使用ImageFZ测量程序冻结百分比（;;上下文测试，R = 0.970线索测试，R = 0.934调理，R = 0.947）呈高度相关性与通过人的观察打进使用光子束为基础的计算机测量系统（1秒10秒，2秒10秒，或Latency3）和人类观察者较低（空调，R = 0.503，0.593，0.761和;上下文吨斯，R = 0.772，0.819和0.912）相比，使用ImageFZ和人的观察（ 图7A和7B）测定的冷冻百分比之间的相关性。此外， 图7表明，通过人的观察获得，并在每只小鼠使用ImageFZ冻结百分比之间的差异是最小的差异。这些结果表明，测定冷冻的量，当使用ImageFZ测定冷冻百分比是相似的，通过人的观察和ImageFZ获得高度准确的。 <img alt="图1" fo:content-width="5in" fo:src="/files/ftp_upload/50871/50871fig1highres.jpg" src="/files/ftp_upload/50871/50871fig1.jpg" /> 图1。设备的背景和线索恐惧条件测试（A）的丙烯酸方腔的调节和背景测试，（ <str翁&gt; B）在一个白色的塑料地板黑，刺鼠，或稀褐家鼠（顶部）和一个黑色的塑料地板小白鼠（底部）electrifiable黑色金属网格金属网格，网格的放大图像显示在右侧面板，（C）白噪声/音频发生器和冲击发生器，（D）一个隔音的房间，和（E）的丙烯酸三角室配备了一台二层为线索的考验。 <a href="https://www.jove.com/files/ftp_upload/50871/50871fig1highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图2" fo:content-width="5in" fo:src="/files/ftp_upload/50871/50871fig2highres.jpg" src="/files/ftp_upload/50871/50871fig2.jpg" /> 图2。上下文的协议的示意图。（一 ）概述和线索恐惧条件测试，（B）调节，（C）C导论测试，以及（D）线索测试。 <a href="https://www.jove.com/files/ftp_upload/50871/50871fig2highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图3" fo:content-width="5in" fo:src="/files/ftp_upload/50871/50871fig3highres.jpg" src="/files/ftp_upload/50871/50871fig3.jpg" /> 图3。由ImageFZ软件程序的图像分析。对于每对连续图像，区域（像素），通过该鼠标移动的量由ImageFZ被计算。当这个区域是低于一定的阈值（ 例如，30个像素），则该行为被判断为“冻结”。当区域的数量等于或超过该阈值，则该行为被认为是“非冻结&#39;。 <a href="https://www.jove.com/files/ftp_upload/50871/50871fig3highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图4" fo:content-width="6in" src="/files/ftp_upload/50871/50871fig4highres.jpg" width="500" /> 图4。来自于使用ImageFZ与那些通过人的观察测量不同的帧速率计算出的图像冻结百分比的比较。的恐惧条件测试使用雄性C57BL/6J小鼠（n = 5）进行。在测试过程中，两名观察员打进冻结行为。同时，实时图像被抓获使用ImageFZ程序4 fps的。在4帧捕获的文件中提取帧对应于1 fps或2帧拍摄的图像后缩小。 “速率（帧/秒）&#39;的参数值设置为1，2，或4帧，并且在每个60秒斌冻结百分比从图像文件使用ImageFZ离线分析计算。每个点代表每个60秒斌冻结百分比。计算从人的观察和ImageFZ分析而获得的数据之间的Pearson相关系数。<a href="https://www.jove.com/files/ftp_upload/50871/50871fig4highres.jpg" target="_blank">点击此处查看大图。</a> <img alt="图5" fo:content-width="6in" src="/files/ftp_upload/50871/50871fig5highres.jpg" width="500" /> 图5。从图像计算使用ImageFZ和那些通过人的观察测量不同凝固条件值冻结百分数比较，恐惧条件测试使用雄性C57BL/6J小鼠（n = 5）进行。在测试过程中，两名观察员记录的冻结行为，以及现场图像利用ImageFZ程序捕获。在每个60秒斌冻结百分比是通过ImageFZ离线分析计算从图像（1帧/秒），设置的&#39;冻结准则（像素）“的参数值到20，30，或40像素。每个点代表每个60秒斌冻结百分比。 Pearson相关合作从人的观察和ImageFZ分析得到的数据之间efficients计算在每个测试。 <a href="https://www.jove.com/files/ftp_upload/50871/50871fig5highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图6" fo:content-width="5in" fo:src="/files/ftp_upload/50871/50871fig6highres.jpg" src="/files/ftp_upload/50871/50871fig6.jpg" /> 图6。冻结的百分比是使用自动系统和人工观察雄性C57BL/6J小鼠无条件和有条件组（n = 5，各组）进行测量 。 （ 一 ）人类的观察，（B）ImageFZ，（三）冻结监控系统1（1秒10秒），（四）冻结监控系统2（2秒10秒），和（E）冻结监控系统3（Latency3）。组比较采用双向重复测量ANOVA，然后进行t检验（执行不合作nditioned组与空调组比较，* P &lt;0.05;†，P &lt;0.01）。使用ImageFZ类似于那些通过人的观察打进获得的数据。 <a href="https://www.jove.com/files/ftp_upload/50871/50871fig6highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图7" fo:content-width="4.3in" src="/files/ftp_upload/50871/50871fig7highres.jpg" width="420" /> 图7。相关和冻结百分比，使用自动系统和人工观察来测定。（AB）的散点图和Pearson相关系数冻结百分比打进通过自动系统和人工观测之间的差异频率分布显示。冻结百分比，使用ImageFZ计算，有高度相关性与通过人的观察所得。小于10％的DIF（CF）出现次数从干扰自动化系统与人类观察得到的冻结之间的百分比是最高的时候使用ImageFZ是与那些通过人的观察分析比较的数据进行分析。 <a href="https://www.jove.com/files/ftp_upload/50871/50871fig7highres.jpg" target="_blank">点击这里查看大图。</a> <table border="0" cellpadding="0" cellspacing="0"><tbody><tr><td></td><td>方差分析</td><td></td><td></td></tr><tr><td></td><td>条件</td><td>时间</td><td>条件x时间</td></tr><tr><td>第1天（空调） </td><td></td><td></td><td></td></tr><tr><td>人的</td><td> F（1,8）= 28.53，P = 0.0007 </td><td> F（7,56）= 20.79，P &lt;0.0001 </td><td> F（7,56）= 16.58，P &lt;0.0001 </td></tr><tr><td> ImageFZ </td><td> F（1,8）= 13.97，P = 0.0057 </td><td> F（7,56）= 21.40，P &lt;0.0001 </td><td> F（7,56）= 11.69，P &lt;0.0001 </td></tr><tr><td>冻结监视器（1sec10sec） </td><td> F（1,8）= 5.16，P = 0.0528 </td><td> F（7,56）= 2.39，P = 0.0329 </td><td> F（7,56）= 0.72，P = 0.6572 </td></tr><tr><td>冻结监视器（2sec10sec） </td><td> F（1,8）= 4.07，P = 0.0782 </td><td> F（7,56）= 3.44，P = 0.0039 </td><td> F（7,56）= 1.52，P = 0.1803 </td></tr><tr><td>冻结监视器（Latency3） </td><td> F（1,8）= 4.44，P = 0.0682 </td><td> F（7,56）= 9.94，P &lt;0.0001 </td><td> F（7,56）= 4.33，P = 0.0007 </td></tr><tr><td>第2天（上下文） </td><td></td><td></td><td></td></tr><tr><td>人的</td><td> F（1,8）= 42.94，P = 0.0002 </td><td> F（4,32）= 1.91，P = 0.1336 </td><td> F（4,32）= 1.48，P = 0.2302 </td></tr><tr><td> ImageFZ </td><td> F（1,8）= 49.61，P = 0.0001 </td><td> F（4,32）= 2.06，P = 0.1087 </td><td> F（4,32）= 0.83，P = 0.5174 </td></tr><tr><td>冻结监视器（1sec10sec） </td><td> F（1,8）= 20.28，P = 0.002 </td><td> F（4,32）= 1.63，P = 0.1918 </td><td> F（4,32）= 0.55，P = 0.6997 </td></tr><tr><td>冻结监视器（2sec10sec） </td><td>F（1,8）= 40.20，P = 0.0002 </td><td> F（4,32）= 2.66，P = 0.0504 </td><td> F（4,32）= 1.20，P = 0.3306 </td></tr><tr><td>冻结监视器（Latency3） </td><td> F（1,8）= 35.30，P = 0.0003 </td><td> F（4,32）= 2.49，P = 0.0626 </td><td> F（4,32）= 1.09，P = 0.3793 </td></tr></tbody></table> 表1中。统计数据的比较。

Watch this Scientific Journal Video about Contextual and Cued Fear Conditioning Test Using a Video Analyzing System in Mice at JoVE.com

Contextual and Cued Fear Conditioning Test Using a Video Analyzing System in Mice

本文介绍了一个协议，用于使用视频分析系统来评估恐惧学习和记忆的小鼠上下文和线索恐惧条件测试。

背景和线索恐惧制约试验小鼠中使用视频分析系统

The contextual and cued fear conditioning test is one of the behavioral tests that assesses the ability of mice to learn and remember an association ...

contextual-cued-fear-conditioning-test-using-video-analyzing-system

Research

JoVE Journal

Behavior

50.9K 次观看.  Fujita Health University. 本文介绍了一个协议，用于使用视频分析系统来评估恐惧学习和记忆的小鼠上下文和线索恐惧条件测试。 

视频: Contextual and Cued Fear Conditioning Test Using a Video Analyzing System in Mice

Trace Fear Conditioning in Mice

In this experiment we present a technique to measure learning and memory. In the trace fear conditioning protocol presented here there are five pairings between a neutral stimulus and an unconditioned stimulus. There is a 20 sec trace period that separates each conditioning trial. On the following day freezing is measured during presentation of the conditioned stimulus (CS) and trace period. On the third day there is an 8 min test to measure contextual memory. The representative results are from mice that were presented with the aversive unconditioned stimulus (shock) compared to mice that received the tone presentations without the unconditioned stimulus. Trace fear conditioning has been successfully used to detect subtle learning and memory deficits and enhancements in mice that are not found with other fear conditioning methods. This type of fear conditioning is believed to be dependent upon connections between the medial prefrontal cortex and the hippocampus. One current controversy is whether this method is believed to be amygdala-independent. Therefore, other fear conditioning testing is needed to examine amygdala-dependent learning and memory effects, such as through the delay fear conditioning.

In the following experiment we describe a protocol for trace fear conditioning in mice. This type of associative memory includes a trace period that separates the neutral stimulus and the unconditioned stimulus.

跟踪恐惧条件小鼠

In this experiment we present a technique to measure learning and memory. In the trace fear conditioning protocol presented here there are five pairings ...

科研

行为学

Using the Threat Probability Task to Assess Anxiety and Fear During Uncertain and Certain Threat

Fear of certain threat and anxiety about uncertain threat are distinct emotions with unique behavioral, cognitive-attentional, and neuroanatomical components. Both anxiety and fear can be studied in the laboratory by measuring the potentiation of the startle reflex. The startle reflex is a defensive reflex that is potentiated when an organism is threatened and the need for defense is high. The startle reflex is assessed via electromyography (EMG) in the orbicularis oculi muscle elicited by brief, intense, bursts of acoustic white noise (i.e., &#8220;startle probes&#8221;). Startle potentiation is calculated as the increase in startle response magnitude during presentation of sets of visual threat cues that signal delivery of mild electric shock relative to sets of matched cues that signal the absence of shock (no-threat cues). In the Threat Probability Task, fear is measured via startle potentiation to high probability (100% cue-contingent shock; certain) threat cues whereas anxiety is measured via startle potentiation to low probability (20% cue-contingent shock; uncertain) threat cues. Measurement of startle potentiation during the Threat Probability Task provides an objective and easily implemented alternative to assessment of negative affect via self-report or other methods (e.g., neuroimaging) that may be inappropriate or impractical for some researchers. Startle potentiation has been studied rigorously in both animals (e.g., rodents, non-human primates) and humans which facilitates animal-to-human translational research. Startle potentiation during certain and uncertain threat provides an objective measure of negative affective and distinct emotional states (fear, anxiety) to use in research on psychopathology, substance use/abuse and broadly in affective science. As such, it has been used extensively by clinical scientists interested in psychopathology etiology and by affective scientists interested in individual differences in emotion.

Potentiation of the startle reflex is measured via electromyography of the orbicularis oculi muscle during low (uncertain) and high (certain) probability electric shock threat in the Threat Probability Task. This provides an objective measure of distinct negative emotional states (fear/anxiety) for research on psychopathology, substance use/abuse, and broad affective science.

使用威胁概率任务，以评估焦虑和恐惧在不确定性和一定的威胁

Fear of certain threat and anxiety about uncertain threat are distinct emotions with unique behavioral, cognitive-attentional, and neuroanatomical ...

Quantifying Social Motivation in Mice Using Operant Conditioning

In this protocol, social motivation is measured in mice through a pair of operant conditioning paradigms. To conduct the experiments, two-chambered shuttle boxes were equipped with two operant levers (left and right) and a food receptacle in one chamber, which was then divided from the second chamber by an automated guillotine door covered by a wire grid. Different stimulus mice, rotated across testing days, served as a social stimulus behind the wire grid, and were only visible following the opening of the guillotine door. Test mice were trained to lever press in order to open the door and gain access to the stimulus partner for 15 sec. The number of lever presses required to obtain the social reward progressively increased on a fixed schedule of 3. Testing sessions ended after test mice stopped lever pressing for 5 consecutive minutes. The last reinforced ratio or breakpoint can be used as a quantitative measure of social motivation. For the second paradigm, test mice were trained to discriminate between left and right lever presses in order to obtain either a food reward or the social reward. Mice were rewarded for every 3 presses of each respective lever. The number of food and social rewards can be compared as a measurement of the value placed upon each reward. The ratio of each reward type can also be compared between mouse strains and the change in this ratio can be monitored within testing sessions to measure satiation with a given reward type. Both of these operant conditioning paradigms are highly useful for the quantification of social motivation in mouse models of autism and other disorders of social behavior.

This article describes a new protocol for the quantitative measurement of social motivation in mice using operant conditioning for a social reward. The protocol is useful for the measurement of social motivation in mouse models of autism and other disorders of social behavior.

量化的社会动机小鼠使用操作条件反射

In this protocol, social motivation is measured in mice through a pair of operant conditioning paradigms. To conduct the experiments, two-chambered ...

Determining Ultrasonic Vocalization Preferences in Mice using a Two-choice Playback Test

Mice emit ultrasonic vocalizations (USVs) during a variety of conditions, such as pup isolation and adult social interactions. These USVs differ with age, sex, condition, and genetic background of the emitting animal. Although many studies have characterized these differences, whether receiver mice can discriminate among objectively different USVs and show preferences for particular sound traits remains to be elucidated. To determine whether mice can discriminate between different characteristics of USVs, a playback experiment was developed recently, in which preference responses of mice to two different USVs could be evaluated in the form of a place preference.
First, USVs from mice were recorded. Then, the recorded USVs were edited, trimmed accordingly, and exported as stereophonic sound files. Next, the USV amplitudes generated by the two ultrasound emitters used in the experiment were adjusted to the same sound pressure level. Nanocrystalline silicon thermo-acoustic emitters were used to play the USVs back. Finally, to investigate the preference of subject mice to selected USVs, pairs of two differing USV signals were played back simultaneously in a two-choice test box. By repeatedly entering a defined zone near an ultrasound emitter and searching the wire mesh in front of the emitter, the mouse reveals its preference for one sound over another. This model allows comparing the attractiveness of the various features of mouse USVs, in various contexts.

Ultrasonic vocalizations (USVs) in mice differ depending on age, sex, condition, and genetic background. Using two ultrasound emitters broadcasting simultaneously in different locations, this two-choice test can evaluate murine recognition and preference responses to different characteristics of USVs.

采用双选择播放测试确定超声波发声首小鼠

Mice emit ultrasonic vocalizations (USVs) during a variety of conditions, such as pup isolation and adult social interactions. These USVs differ with age, ...

Whisker-signaled Eyeblink Classical Conditioning in Head-fixed Mice

Eyeblink conditioning is a common paradigm for investigating the neural mechanisms underlying learning and memory. To better utilize the extensive repertoire of scientific techniques available to study learning and memory at the cellular level, it is ideal to have a stable cranial platform. Because mice do not readily tolerate restraint, they are usually trained while moving about freely in a chamber. Conditioned stimulus (CS) and unconditioned stimulus (US) information are delivered and eyeblink responses recorded via a tether connected to the mouse's head. In the head-fixed apparatus presented here, mice are allowed to run as they desire while their heads are secured to facilitate experimentation. Reliable conditioning of the eyeblink response is obtained with this training apparatus, which allows for the delivery of whisker stimulation as the CS, a periorbital electrical shock as the US, and analysis of electromyographic (EMG) activity from the eyelid to detect blink responses.

The preparation presented here for whisker-signaled eyeblink conditioning in head-fixed mice precisely stimulates specific whiskers while allowing mice to ambulate on a cylindrical treadmill. A whisker stimulation conditioned stimulus (CS) paired with a periorbital shock unconditioned stimulus (US) results in reliable associative learning on this apparatus.

在头固定小鼠胡须信号眨眼经典条件反射

Eyeblink conditioning is a common paradigm for investigating the neural mechanisms underlying learning and memory. To better utilize the extensive ...

Reducing State Anxiety Using Working Memory Maintenance

The purpose of this protocol is to explain how to examine the relationship between working memory processes and anxiety by combining the Sternberg Working Memory (WM) and the threat of shock paradigms. In the Sternberg WM paradigm, subjects are required to maintain a series of letters in the WM for a brief interval and respond by identifying whether the position of a given letter in the series matches a numerical prompt. In the threat of shock paradigm, subjects are exposed to alternating blocks where they are either at risk of receiving unpredictable presentations of a mild electric shock or are safe from the shock. Anxiety is probed throughout the safe and threat blocks using the acoustic startle reflex, which is potentiated under threat (Anxiety-Potentiated Startle (APS)). By conducting the Sternberg WM paradigm during the threat of shock and probing the startle response during either the WM maintenance interval or the intertrial interval, it is possible to determine the effect of WM maintenance on APS.

This protocol demonstrates how to measure anxiety-potentiated startle during the Sternberg Working Memory paradigm.

使用工作记忆维护降低状态焦虑

The purpose of this protocol is to explain how to examine the relationship between working memory processes and anxiety by combining the Sternberg Working ...

Using Electroencephalography Measurements and High-quality Video Recording for Analyzing Visual Perception of Media Content

This article explores a method to detect differences in visual perception in humans. The method used is based on the psychological (or "cognitive") function of eyeblinks. Participants' eyeblinks are detected and acquired while watching videos specifically created for the investigation. The detection and acquisition of eyeblinks are carried out with the help of a 20-channel electroencephalographic (EEG) wireless device. The international 10-20 system for electrode placement is followed. A high-definition (HD) video camera is used to record participants' facial expressions, for contrast purposes. Instead of using pre-existing media content, purpose-made video content has been created following specific criteria of interest for this investigation, with stimuli enabling researchers to manage the precise parameters of interest. Otherwise, results could be contaminated with uncontrolled variables. The synchronization of the presentation of video stimuli with EEG recordings needs to be done in milliseconds. Analysis of collected data is performed with robust software for working with big matrices. Statistically significant differences in eyeblink rate related to media professionalization and editing style are found with the reported experimental procedures.

We present detection, acquisition, and analysis of eyeblink rates while watching media content.

利用脑电图测量和高质量视频记录分析媒体内容的视觉感知

This article explores a method to detect differences in visual perception in humans. The method used is based on the psychological (or "cognitive") ...

Analyzing Spatial Learning and Prosocial Behavior in Mice Using the Barnes Maze and Damsel-in-Distress Paradigms

The Barnes maze is a reliable measure of spatial learning and memory that does not require food restriction or exposure to extremely stressful stimuli. The Barnes maze can also assess other mouse behaviors, such as general motivation to escape from the maze platform and exploratory behavior. The Barnes maze can measure whether a genetic mutation or environmental variable can impact the acquisition and retention of spatial memories, as well as provide information about the search strategy employed by the mice. Here we use the Barnes maze to detect a memory deficit in adult mice following a single developmental ethanol exposure event. The newly described Damsel-in-Distress paradigm exposes a male mouse to a female mouse trapped in a chamber in the open center field of the arena. It provides an opportunity for the mouse to socially respond to the trapped female and exhibit prosocial behavior. The Damsel-in-Distress paradigm can also be used to examine mouse behavior in a novel arena and measure locomotor activity. Both the Barnes Maze and the Damsel-in-Distress protocols require minimal financial investment and most aspects of the tests can be constructed from common lab supplies. These flexible and accessible tools can also be used to detect behavioral changes over the course of development.

This protocol measures spatial learning and memory using the Barnes maze. A novel Damsel-in-Distress paradigm is used to assess locomotor activity and prosocial behavior in mice.

利用巴恩斯迷宫和困境中的破坏范式分析小鼠的空间学习和社会行为

The Barnes maze is a reliable measure of spatial learning and memory that does not require food restriction or exposure to extremely stressful stimuli. ...

Visual Classical Conditioning in Wood Ants

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, studies implementing classical conditioning paradigms with harnessed insects have been important for investigating the exact cues that individuals learn and the neural mechanisms underlying learning and memory formation. Here we present a protocol for evoking visual associative learning in wood ants through classical conditioning. In this paradigm, ants are harnessed and presented with a visual cue (a blue cardboard), the conditional stimulus (CS), paired with an appetitive sugar reward, the unconditional stimulus (US). Ants perform a Maxilla-Labium Extension Reflex (MaLER), the unconditional response (UR), which can be used as a readout for learning. Training consists of 10 trials, separated by a 5-minute intertrial interval (ITI). Ants are also tested for memory retention 10 minutes or 1 hour after training. This protocol has the potential to allow researchers to analyze, in a precise and controlled manner, the details of visual memory formation and the neural basis of learning and memory formation in wood ants.

We present a protocol for the classical conditioning of harnessed ants that permits researchers to study visual learning and memory formation at a level of analysis not possible with freely moving individuals.

木蚁视觉经典调理

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, ...

Fear Incubation Using an Extended Fear-Conditioning Protocol for Rats

Emotional memory has been primarily studied with fear-conditioning paradigms. Fear conditioning is a form of learning through which individuals learn the relationships between aversive events and otherwise neutral stimuli. The most-widely utilized procedures for studying emotional memories entail fear conditioning in rats. In these tasks, the unconditioned stimulus (US) is a footshock presented once or several times across single or several sessions, and the conditioned response (CR) is freezing. In a version of these procedures, called cued fear conditioning, a tone (conditioned stimulus, CS) is paired with footshocks (US) during the training phase. During the first test, animals are exposed to the same context in which training took place, and freezing responses are tested in the absence of footshocks and tones (i.e., a context test). During the second test, freezing is measured when the context is changed (e.g., by manipulating the smell and walls of the experimental chamber) and the tone is presented in the absence of footshocks (i.e., a cue test). Most cued fear conditioning procedures entail few tone-shock pairings (e.g., 1-3 trials in a single session). There is a growing interest in less common versions involving an extensive number of pairings (i.e., overtraining) related to the long-lasting effect called fear incubation (i.e., fear responses increase over time without further exposure to aversive events or conditioned stimuli). Extended fear-conditioning tasks have been key to the understanding of fear incubation&#8217;s behavioral and neurobiological aspects, including its relationship with other psychological phenomena (e.g., post-traumatic stress disorder). Here, we describe an extended fear-conditioning protocol that produces overtraining and fear incubation in rats. This protocol entails a single training session with 25 tone-shock pairings (i.e., overtraining) and a comparison of conditioned freezing responses during context and cue tests 48 h (short-term) and 6 weeks (long-term) after training.

We describe an extended fear-conditioning protocol that produces overtraining and fear incubation in rats. This protocol entails a single training session with 25 tone-shock pairings (i.e., overtraining) and a comparison of conditioned freezing responses during context and cue tests 48 h (short-term) and 6 weeks (long-term) after training.

对大鼠使用扩展的恐惧调节协议进行恐惧孵化

Emotional memory has been primarily studied with fear-conditioning paradigms. Fear conditioning is a form of learning through which individuals learn the ...