这项工作是由美国国防部（W81XWH-13-01-0243）和美国国立卫生研究院（R37 AG008796）的资助。我们感谢艾伦·贝克在美国西北大学的机械加工车间建设头固定筒设备。我们感谢Shoai服部博士，他在MATLAB和Solidworks指导。我们感谢约翰功率博士对于控制实验中的LabVIEW软件。

所有涉及小鼠程序按照批准的西北大学的机构动物护理和使用委员会基于美国国立卫生研究院颁布了准则的协议进行。 1.缸（图2A） <ol><li>如由Chettih 等人和Heiney 等人描述构建圆柱体。从长泡沫圆柱体14-15。切割滚筒的10cm的长度，并通过中心钻一个孔，以适应轴，直径金属杆12.7毫米（0.5英寸）。如Heiney 等或其他支持（ 如有机玻璃）15安装说明与轴筒上的金属光学平台。 </li><li>竖立在缸体的两侧12.7毫米直径的两个垂直金属棒。 </li><li>连接两个直角坐骑到金属棒。 </li><li>通过直角支架固定两个棒（5厘米长12.7毫米直径）。第斜角这些棒电子结束，以上等级的液压缸（ 图2G）的中心的端部。 </li><li>钻和挖掘一个洞杆的两端拉平，以适应，将举行连接器的翅膀到位（ 图2G）二4-40机器螺钉。 </li></ol> 2.装配晶须激励系统（图2B） <ol><li>从一个普通梳子梳剪一个10齿段。 </li><li>切缝大约2-3毫米宽，大约5毫米深的梳接受的2层为矩形的弯曲致动器的顶部。 </li><li>焊接一个导线的弯曲致动器的每个表面。使用由制造商提供的焊料和焊剂。覆盖致动器和焊料用绝缘带，以保护鼠标和用户从电流。 </li><li>滑致动器条插入切入梳狭缝，使得弯曲致动器条位于垂直于梳的平面。角梳，以便它在4倾斜5°，更自然跟随鼠标的脸的曲线。 </li><li>固定致动器带材的端部到与速凝环氧树脂的梳子的顶部。 注:钎焊到弯曲致动器带的线应连接到提供40伏至致动器的变阻器。以前的实验已经表明，40伏是有效的调理，但不引起任何惊恐反应。 </li><li>放置弯曲致动器和一个可移动的安装梳。连接到作为通常用于体外电生理学钻机一个磁性底座的移液管固定器效果很好。 </li></ol> 3.装配的坐骑杆汽缸上方的连接器（图2C，2E） 注意:所述的连接器是一个三维打印来自用于由Weiss和Disterhoft和加尔维斯等系绳的安费诺221系列尼龙带建模7孔条10,17。 <ol><li>轻按条第一洞为0-80 X-1"机器螺钉，这将作为一个锁紧螺钉锚头状的连接器。贴上一个0-80螺母锁紧螺钉与氰基丙烯酸胶的头部，以便于用手转动螺杆。 </li><li>留一个孔的螺纹孔后，空（为了留出空间0-80机螺母），并通过剩余的五杆洞推五枚镀金瑞莱-TAC插槽。 </li><li>剥离涂层脱落五龙线（从以太网电缆获得的电线）和焊料插座的两端。 注意:第一两根导线将被用于记录肌电图（EMG）信号，将检测以下从放大器信号的滤波和放大的眨眼反应。后两个将从刺激隔离器发出的电流信号。最后线将作为接地连接。这些导线将连接到用于定时的CS-US递送系统。 </li><li>确保导线和插座用MULT之间的电连接imeter。穿上连续性设置万用表并保持探针销和其他探头的导线的裸露端部中的一个。销与导线电连接时的电阻低，万用表发出声调。 <ol><li>另外，如果万用表不具有连续性设置，测量销与导线之间的电阻。有，如果有测定的电阻很小的电连续性。 </li></ol></li><li>从两个TO-220式晶体管制备连接器的翅膀。切断晶体管的发射极，基极，和集电极引线和表面平整，因此它可以被固定至连接器。 </li><li>切安装卡的边缘违反孔，从而标签可以滑的螺钉的头部的下方。 </li><li>环氧树脂的晶体管的连接器。在安装片的孔将被用于将连接器固定到汽缸上方的杆。 </li><li>适用的环氧导线的PR基从插座otruding并允许环氧干燥。环氧绝缘会和保护连接。 </li></ol> 4.准备头饰（图2D，2E） 注意:头盔带为三维打印来自用于由Weiss和Disterhoft和加尔韦斯等人 10,17的头盔的安费诺221系列尼龙带建模7孔条。这片已经不再生产的商业化。打印机文件可以从该杂志的网站上下载。 <ol><li>点击第一个洞带为0-80×1"机螺钉，并留下一洞的第一洞后是空的。 </li><li>通过剩余的五杆洞的底部推五枚镀金插针（通过较窄的结束）。使用老虎钳，方便推销插入带均匀。 </li><li>使用热汽提器（0.005英寸内径）除去约0.5cm聚酰亚胺涂层的关闭的不锈钢丝的端部，焊接W的剥离端愤怒的销中的一个的打开和切断丝0.6-0.7厘米。 </li><li>剥去电线的端部约0.2厘米到允许电流传递给动物。 </li><li>重复步骤4.3-4.4其余三个电线。 注:前两线将记录EMG反应，后两个将用于发送电击动物。 </li><li>作为最后的（第五）的电线将作为地线，切成约5厘米的未涂覆的不锈钢丝（0.005英寸直径），以及焊料到其余针。 </li><li>确保电线用万用表测量引脚之间的连续性。 （在地线，这是未涂覆的情况下，该万用表探针可以在任何地方沿着电线放置）。 </li></ol> 5.手术准备（图2F） <ol><li>消毒所有手术工具，包括头盔。通过在酒精中浸泡，然后用无菌盐水冲洗消毒头盔。 </li><li>麻醉鼠标在3-4％V感应室aporized异氟烷1-2 L到分钟的氧气的流量进行混合。 </li><li>确定该动物是用脚趾捏完全麻醉。鼠标是完全麻醉时，它不向脚趾捏本能反应。 </li><li>一旦动物已被完全麻醉，注入剂量盐酸丁丙诺啡作为镇痛（0.05-2毫克/千克，SC），剃动物的头的顶部，并将其放置在覆盖加热垫立体框架的基础上，确定该动物被完全麻醉，头部固定到框架上。保持汽化的异氟烷流向动物，以1-2 L到分钟的氧气的流量切换到2％。适用眼药膏少量的角膜。 </li><li>消毒用聚维酮碘和醇三个时间每个头皮，这两者之间交替。 </li><li>以数字10或15的手术刀刀片，使沿头皮的中线切口，从T的前面露出颅骨他的眼睛过去interparietal骨（〜1.5-2厘米）。 </li><li>憋微剪辑的皮肤皮瓣。放置一个剪辑的眼睛上方一沿延髓-尾轴的中部，并且一个在头骨的背面两侧（ 即，可以引用六个剪辑所有）。为了防止头盔撕脱，暴露尽可能多的头骨作为可能的，包括侧面和背面。这将增加表面积为粘合剂粘接水泥的应用。 </li><li>使用手术刀，沿头骨移除骨膜和确保一个干净，干燥的工作表面的顶部刮去。清洁用3％的过氧化氢三次颅骨的顶部。 </li><li>钻两个孔入颅，大小34倒锥毛刺或1.6毫米雕刻铣刀钻头接受00-90螺丝（在0.0625。长）。螺丝将提供电接地连接。置于前囟的前面有一个孔，而另一个在LAMBDA的前，左中线时调理右击Ť眼球。放在每孔一个螺丝。降低螺杆0.28毫米入颅每个整圈;两圈就足够了。 </li><li>为了增加对水泥的表面积和握力创建的在头骨上的直径大约0.75毫米几个小草皮。 </li><li>取完成头盔和缠绕地线在周围的两个螺钉数字8的配置。允许一些松弛（〜了头盔和螺杆〜1.5厘米）中的地线，以便在头盔能够适当稍后被定位。 注意:八字形通常保证了导线和接地螺钉之间的良好的电连接。导线也可焊接到螺钉，以确保电连接。 </li><li>涂抹粘合剂粘接水泥。遵循制造商的说明用于混合粘合剂粘接水泥或在寒冷的陶瓷混合菜混合4勺的L-粉混合，碱的8滴，和2滴催化剂的（保证温度条Òn中的混合菜是全黑的。）的低温扩展了水泥的工作时间。 <ol><li>大衣颅骨和宽松的粘接水泥螺钉，使水泥干燥。这应该只需要几分钟的时间。 </li></ol></li><li>水泥干燥后，垂直地定位的头盔，与销站立，颅骨上方。保持在适当位置的头盔具有类似于连接器的支架 注:请参阅"3.组装连接器"-The持有人只需要与镀金插座一个五孔带，以获得头盔的引脚。保持器安装于立体定位框架上的臂。使用该保持架和臂的方便了头盔和导线的定位。 </li><li>头状被定位后，剥离背部周围的眶周区域的皮肤和定位在两个冲击的电线，以允许0.2厘米剥离端休息的皮肤下方和大约2-4毫米直接尾椎EY即不允许两导线的端部相互接触。在肌层眼轮匝肌两肌线的位置在眼​​窝上方。 <ol><li>如果需要的话，切断导线的端部，如果他们是太长，看起来好像它们可以直接划伤轨道，并导致感染。如果切断电线的端部，确保有暴露足够裸线。另外，如果导线似乎太长，弯曲线向后在基地，在那里他们伸出头盔的。 </li><li>水泥的电线的基部（ 即 ，头状接近结束）在与所述粘合剂粘接水泥的小扩散颅骨地方并晾干（使用半先前描述的部分）。 </li></ol></li><li>取出微型夹，轻轻折皮肤皮瓣又回到了水泥。让皮肤自然沉降，以防止在皮肤上的任何部分的张力，以避免眼睑失真，防止眨眼反应，和痛苦的动物。 </li><li>用牙科水泥密封暴露面积，涵盖了从皮肤切与水泥的头盔的顶部的边缘。要特别小心，以免对眼睛或头盔的引脚淋漓水泥。部分固化的水泥可以平滑并与被浸有牙科用粘固剂溶剂棉签操纵。让水泥卸下头盔架之前完全干燥。 </li><li>让动物从麻醉在温暖的表面取代动物放回笼子前恢复。辖Metacam（1毫克/千克，SQ），然后将动物放回其笼子。 </li><li>测试或训练前让动物恢复5-7天。根据你的机构的指导方针保持标准的术后护理的动物。它已经从手术恢复后的动物应检查每天至少一次。当心不良的饮食和/或饮酒和无精打采行为的迹象。如果疼痛之嫌，提供Metacam每24小时（相同剂量在手术后的结束），直至缓解。利多卡因可以局部施加到如果动物划伤或示出的不适的迹象伤口。 </li></ol> 6.将鼠标上的气缸和培训（图2G） <ol><li>鼠标放置到汽缸，通过由尾巴用一只手握住，然后把持用食指和拇指与其他肩部后面鼠标抑制用一只手简单地将鼠标。缠绕在鼠标的腹部和躯干其余手指。 </li><li>用手制止没有鼠标，连接器连接到鼠标的头部的头盔，并打开锁紧螺钉。 </li><li>轻轻地放置在汽缸鼠标举行到位而连接连接到框架上。使用这两个螺钉将连接器固定到气缸上方的棒。 </li><li>一旦连接器已被固定在气缸支架，RELEASE鼠标上的搁置。给鼠标的习惯与气缸两会的一天。让每个习惯会话持续时间相同，因为每个空调会话。 <ol><li>习惯化期间记录自发闪烁速率和预暴露小鼠晶须振动的CS，以最小化的惊恐反应的发生。应用CS作为实际调理会话期间，但没有触电美国。 </li><li>通过将鼠标的压电系统关闭（约0.5厘米）到经调节的一侧（右侧），而滑动梳过个别晶须的齿申请晶须刺激的CS。确保相同晶须通过将压电系统在相同的位置，从每天刺激每天。 </li></ol></li><li>开始训练鼠标放在经典条件。提供每天训练两会五天。离开每次训练大约两个小时。 <ol><li>对空调组，熟食店每个版本具有100毫秒长电脉冲美国配对的250毫秒长胡须振动CS（60赫兹）的会议30试验（0.4 mA数 - 3毫安，从WPI A385R刺激隔离器送到）。分隔CS和美国以250毫秒长无刺激扫描间隔以使任务依赖于海马（ 图1）23。 </li><li>调整电击的强度，每个动物，这样一眨被诱发。独立审判与30-60秒的随机intertrial间隔（平均45秒，每节30分钟，总时间的。） </li><li>单独递送30不成对CS和美国单独试验的每个（为一个总共有64试验）向​​pseudoconditioned组，用同样的刺激参数如所述的空调组。确保仅在CS和美国单独试验正在pseudorandomized使得无刺激呈现两次以上在一排。分隔每个试验以22.5秒的平均intertrial间隔。 </li></ol></li><li>德在为了在整个习惯和训练65分贝肝脏背景噪声掩盖由晶须刺激产生的振动的任何噪声。 </li></ol>

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</ol>

作者宣称，他们没有竞争的经济利益。

经典眨眼调理联想学习有助于理解基础的学习和记忆的神经基板的有用工具的一种形式。如小鼠在啮齿类眨眼调理采用以前的方法涉及，允许用于动物至约自由移动的腔室。用于在小鼠中眨眼调理的磁头固定制剂，使用在西格尔等人在光诱发痕量眨眼调理由Chettih 等人和Heiney 等人 ，最近利用所描述的装置提供了几个优点，如允许某些眨眼调理该没有可能或在过去已被限制（ 例如， 在体内双光子钙成像）14-16期间要执行的程序和实验。这种类型的制剂也便于使用的其他技术，例如在体内与四极管阵列电。小鼠出现成为distre从约束ssed，该磁头固定制剂的开发是为了保持头的安全，同时减少约束所需的量。虽然这个准备使小鼠走动在圆柱跑步机，其他人也成功收集在一个稳定的平台，从24头固定小鼠行为数据。 然而也有，这与使用头部固定筒系统的出现某些问题。其中值得关注的是，当一个地方开始未受过训练的鼠标放在缸，它可能需要一些时间来学习休息，散步，或在新的设备上运行。大多数老鼠很快就学会休息，散步，或在气缸舒适运行。两个习惯会帮助习惯于小鼠到气缸和第二届中部，大多数鼠标舒适的圆柱体。出现的另一个问题是头盔撕脱。目前已在多在白天训练课过去的一些问题，这一点，但使用胶粘剂粘接的c键相已经改善与头盔撕脱的问题。使用胶粘剂粘接水泥，因为使用这种技术已导致50多个手术0％头盔撕脱。有了正确的使用和对颅骨水泥充分传播，研究者应该遇到头盔撕脱没有问题。最后，通过在习惯会话呈现在CS，可能有大约潜在抑制一些关注。很清楚，但是，将小鼠成功空调（ 见图7）。注意，晶须振动的强度可通过改变电压的量由变阻器被传递进行修改。如果小鼠无法了解，电压可提高到支持成功调理，同时也最大限度地减少惊恐反应。研究人员还可以改变被输送到动物电击美国的类型。的电击美国可以是一个单一的电脉冲持续100毫秒如这里报告。或者，如第在此方案采用Ë刺激隔离器能够发送双相脉冲，研究人员还可以采用120赫兹，每脉冲1毫秒6双相脉冲对一个有效的电击我们。 Boele 等人表示，某些"条件反射"可能不惊人死不休（阿尔法）出现的CS或短潜伏期反应（SLR）25反应的可能性。阿尔法反应被认为是CS发病后一个50毫秒的等待时间内快速眼睑闭合。老鼠经常保持自己的眼睛闭上了审判的期限，按照本阿尔法的回应。单反相机，另一方面，有来自CS发病50-70毫秒的延迟，只有少数的CS-US配对后通常产生。短潜伏期α和单反相机和单反相机可以只有少数的CS-US配对后出现的事实的发病表明它们可能不是由海马小脑电路驱动。这些意见被Boele 等回顾:Therefore求这里记录的CR是否学到响应的问题。 如图7C和7D示出，既没有阿尔法反应也不单反占多数记录的CR的，表明自适应的CR是由前脑和小脑介导的。在图6中其显示延迟到以下的CS发病眨眼响应峰值直方图还表明，虽然某些试验可包括早期发病α或短潜伏期反应，用更多的训练试验中，眨眼反应的峰值实际上发生以后和接近美国的发作，指示适时条件反应的发展。如前所述，晶须刺激和冲击强度分别衰减，这是不令人吃惊的或过于厌恶，但仍有效地产生自适应空调响应的水平。最后， 图7B示出，在CR的尺寸（测量为下的面积在整流和综合的反应，如在图5的曲线）是空调组中比在pseudoconditioned组较大一旦空调组达到了60％的学习标准，表明这些反应需要几个会话学习，不像单反相机和阿尔法响应。 Boele 等人还指出，肌电图测量，而在较大的动物如兔眨眼检测的精确方法，都没有在小型啮齿动物如小鼠25作为实用。他们建议使用磁距离测量技术（MDMT），以避免EMG记录可能表明假阳性信号的检测。的MDMT技术，而在它的灵敏度和眨眼检测的质量令人印象深刻，也呈现具有训练每天麻醉动物，以附加芯片26的缺点。这本身可能混淆学习率。我们发现，肌电信号记录有这里描述的方法很容易获得高时间分辨率，可靠而且比较容易测量和分析的。 图4示出一个代表性的EMG信号检测从训练期间放置在靠近鼠标的眼睛的红外反射光学传感器与相应的信号闪烁。有一个在闪烁的肌电信号和光传感器之间的检测，使用表示肌电图记录眨眼检测的精度明显的相关性。肌电图记录的一个优点是，它允许为最大的时间分辨率。虽然分辨率是通过集成在一个10毫秒时间常数劣化，还可以分析原始肌电图数据，以检测眨眼活性。穗计数是可用于检测的CR 18一个参数。与肌电图记录的一个缺点是信号无疑将通过电工件从电击美国未来被污染（见<s仲&gt;图3 - 5）。然而，这并不妨碍记录空调眨眼反应， 也就是说，那些发作美国之前发生的能力。在采用肌电图记录的另一个缺点是，使用的CR检测本准则，嘈杂的基线可能隐藏否则可能已经检测什么作为CR值。 这里所描述的头固定制剂是类似于由Heiney 等人提出。15有，但是，从它们的优雅系统的某些显着的差别。例如，该方法在此描述用于记录眨眼反应是电线皮下置于眼睛上方以记录EMG活动。这个技术允许眼睑活动的可靠，稳定的记录，因此，该眨眼反应。这些电线的适当放置和固定，确保质量的录音是持续至少两周，实验的持续时间。使用EMG线，而不是一个高的优点13759 Heiney 等 -speed相机是肌电图记录有极高时间分辨率，并且不需要日常定 ​​位和校准，一个相机要求15。相机确实，但是，报价眼睑闭合的直接的视觉决心。 两个系统之间的另一区别是用于CS递送方法。这种特殊的头部固定鼠标准备利用晶须刺激的条件刺激。结果表明，小鼠可以有效地与晶须刺激在CS为条件，正如头固定家兔可以与晶须振动27进行调节。 Heiney 等人通过将空气的弱抽吸到晶须垫15展示了晶须垫作为有效的CS刺激。虽然这两种技术演示有效调节，将梳子在选择胡须和振动梳允许以激发个人whiske的能力r行甚至个别晶须。这种技术已在以前的研究中使用，以允许动物用作其自身的对照（ 即，受激晶须行了比较未刺激晶须的行）20。 总之，对于晶须信号眨眼调理头固定制剂允许用于执行高级的技术和实验了以前是不可能或很难执行一个安全的颅平台。温和晶须刺激用作条件刺激和温和的电击用作条件刺激。眨眼反应被记录皮下放置在眼睑后面的电线。可靠学习表现在与选定的晶须在CS和眶周触电，因为美国的直接刺激小鼠，并没有学习在于给予CS和美国的随机演示小鼠是显而易见的。肌电图录音提供了一个可靠的和rel记录眨眼反应和观察反应跨越培训课程如何改变atively简单的方法。

眨眼调理巴甫洛夫条件反射的一种形式，调查联想学习和记忆的神经机制的模型系统。它已在各种物种，包括人类，兔，猫，大鼠和小鼠中进行了研究。该模式包括两个配对的刺激的呈现:中性条件刺激（CS; 例如 ，一个音，一道闪光，或晶须刺激），以及一个突出的非条件刺激（US; 例如 ，吹气的眼睛，或眶周震）。美国引发的无条件的，反身眨眼反应（ 即 ，UR）。最终，经过配对的CS-美国的一些演讲，主题学习到CS与美国相关联。这个学习表现为一个条件反应（CR），仅由在CS在先于美国的演示引起一个眨眼的形式。 在跟踪的形式眨眼调理包括一些H的无刺激间隔undred毫秒分隔CS和美国（ 图1）。跟踪空调是声明学习的一种形式，因为它需要刺激偶然性1意识。颞间隙要求动物保持CS的神经&#39;跟踪&#39;前脑区域如海马，以便美国和CS成为相关1-6。随着前脑区域，痕迹条件也依赖于小脑7。 眨眼调理，因此，对于存储器的多个方面，包括采集，合并，和检索的调查一个有用的范例。期间眨眼空调，动物的对照组呈现不成对的刺激以随机顺序来测试pseudoconditioning或致敏反应到CS可以由美国介绍单独而不是学习的CS-US关联而引起的。 常用的appara土族的眨眼调理的啮齿类动物的调查是其中啮齿动物被允许在训练过程中8-10自由行动室。与这种类型的装置中，系绳通常附连到被固定在啮齿动物的头骨头饰。系绳允许与美国（有时在CS）的递送和用于发送动物的对这些刺激（ 即眨眼反应）10的响应。系绳本身可以基于刺激的类型来修改递送和眨眼响应如何被记录。 之所以使用"自由移动"拴小鼠眨眼条件是，对小鼠克制斗争。虽然其他物种可能是更适合的约束，在眨眼调理实验使用的小鼠的主要优点是，大多数可用的遗传修饰的突变体菌株的是小鼠品系。除了挣扎，完整的资源小鼠traint导致急性窘迫。最大限度地减少应力的头固定鼠标准备将增加可眨眼调理过程中获得的生理信息。例如，该系统将允许用2-光子显微镜11的皮层神经元的成像。 头固定制剂已在以前的实验用于通过可拆卸颅植入的皮质光学成像被使用， 在啮齿类动物的脑与四极管阵列，双光子钙成像的体内电生理记录，也可作为在小鼠中11为眨眼调理的平台-16。 在头部固定系统，可靠的刺激和录音，确保没有鼠标（ 图2）完全克制。像中的自由移动系统中使用的一项所述的头盔固定到小鼠的颅骨。在训练期间，头状被固定到被连接到棒上的连接器为了一圆柱形的跑步机，以稳定的啮齿动物的头部（ 图2A）。圆柱形跑步机允许鼠标舒适休息，但如果鼠标愿意，也允许其运行或行走。与使用该系统，小鼠可以用晶须振动作为CS和温和的眶周触电，因为美国的（ 图1）进行培训。美国正在通过导线手术放置在皮肤侧至眼睛下方输送。在CS经由附加到一个2层为矩形的弯曲致动器（ 图2B）的梳递送。梳子和弯曲致动器，然后附着到正在训练期间移动到适当的位置，并调整为每个单独的动物的最佳递送的磁性底座。梳子被定位成跨在所选晶须。期间交付的CS，一个信号被发送到弯曲致动器位移的梳子和导致晶须17的振动。 16,18,19。究其原因晶须刺激选择了CS这个实验范式是鼠类动物的勘探过程中他们的触须对躯体感觉信息输入的依赖。晶须刺激已被证明是可靠的和有效的CS 20。此外，鉴于触须系统（ 即 ，桶皮质），晶须刺激的成熟和有组织的皮质衬底作为CS提供映射与学习眨眼空调20,21相关的皮层变化和可塑性优雅的工具。头部固定系统允许对选定的晶须的精确刺激比较刺激神经元和非刺激晶须接收输入神经元之间的反应。最后，小鼠多株表现出与年龄相关的听力损失是相对年轻的成年人<su在空调闪烁期间P&gt; 22，且眼睑闭合改变一个可视化的CS（虽然视觉CS确实改善问题与惊跳反应16）。晶须刺激不受任一这些并发症。 这里介绍的是在对方头部固定筹备眨眼空调，包括CS和美国交付方式独特而重要的修改，并收购了眨眼反应。此装置和在眨眼调理训练模式的可靠性通过学习从空调小鼠和从pseudoconditioned对照动物（ 图7A）的相对平坦的学习曲线的曲线证明。

<table><tbody><tr><td>Exervo TeraNova Foam Roller 36&quot; x 6&quot;&amp;nbsp;</td><td>Amazon</td><td>B002ONUM0E</td><td>For cylinder</td></tr><tr><td>Plexiglas</td><td></td><td></td><td>Custom-made; 1 cm thick</td></tr><tr><td>Metal Rods (12.7&amp;nbsp;mm diameter)</td><td></td><td></td><td>Custom-made</td></tr><tr><td>4-40 machine screw (.25&amp;nbsp;in. long)</td><td>Amazon Supply</td><td>B00F33Q8QO</td><td>For cylinder</td></tr><tr><td>Classic Design Hair Comb</td><td>Conair</td><td>93505WG-320</td><td>For whisker stimulation</td></tr><tr><td>2-Layer Rectangular Bending Actuator</td><td>Piezo Systems</td><td>T220-A4-303X&amp;nbsp;</td><td>For whisker stimulation</td></tr><tr><td>Solder and Flux Kit</td><td>Piezo Systems</td><td>MSF-003-NI</td><td>For whisker stimulation</td></tr><tr><td>Magnetic Base</td><td>Thor Labs</td><td>MB175</td><td>For whisker stimulation</td></tr><tr><td>Threaded rod for magnetic base</td><td></td><td></td><td>Custom-made</td></tr><tr><td>Strips based on 221 series nylon strip connectors from Electronic Connector Corp.</td><td></td><td></td><td>Custom-made, based on Weiss and Disterhoft, 2008</td></tr><tr><td>TO-220 Style Transistor</td><td>Amazon Supply</td><td>B0002ZPZYO&amp;nbsp;</td><td>For connector; for the wings</td></tr><tr><td>Relia-Tac Sockets</td><td>Electronic Connector Corp.</td><td>220-S02</td><td>For connector</td></tr><tr><td>Relia-Tac Pins</td><td>Electronic Connector Corp.</td><td>220-P02</td><td>For headpiece</td></tr><tr><td>0-80 stainless steel machine screw (1 in. long)</td><td>Amazon Supply</td><td>B000FN68EE</td><td>Locking Screw</td></tr><tr><td>0-80 stainless steel machine screw hex nut (5/32 in. thick)</td><td>Amazon Supply</td><td>B000N2TK7Y</td><td>Locking Screw Head</td></tr><tr><td>Loctite Super Glue-Liquid</td><td>Loctite</td><td>1365896</td><td>Cyanoacrylic glue; for the locking screw</td></tr><tr><td>Quick Setting Epoxy</td><td>Ace Hardware</td><td>18613</td><td>For connector and whisker stimulation system</td></tr><tr><td>Ethernet Cable Wires</td><td></td><td></td><td>Ethernet cable can be taken apart to use the individual wires for the connector</td></tr><tr><td>Polyimide coated stainless steel wires (2&amp;nbsp;in. long, .005 in. diameter)</td><td>PlasticsOne</td><td>005sw/2.0 37365 S-S&amp;nbsp;</td><td>For headpiece, EMG and shock wires</td></tr><tr><td>Stainless steel uncoated wire (.005&amp;nbsp;in. diameter)</td><td>AM Systems</td><td>792800</td><td>For headpiece, ground wires</td></tr><tr><td>Tenma Variable Autotransformer</td><td>Tenma</td><td>72-110</td><td>For the whisker stimulation; rheostat to adjust current to the bending actuator</td></tr><tr><td>Amplifier</td><td>A-M Systems</td><td>1700</td><td>Amplifier for filtering and amplifying EMG signals</td></tr><tr><td>WPI A385R stimulus isolator</td><td>World Precision Instruments</td><td>31405</td><td>For the electrical shock</td></tr><tr><td>Isothesia (Isoflurane)</td><td>Henry Schein: Animal Health</td><td>50031</td><td>For surgery; anesthesia</td></tr><tr><td>Buprenex Injectable CIII</td><td>Reckett Benckiser Pharmaceuticals Inc</td><td>NDC 12496-0757-1</td><td>For surgery; analgesic</td></tr><tr><td>Akwa Tears: Lubricant Ophthalmic Ointment&amp;nbsp;</td><td>Akorn</td><td>NDC 17478-062-35</td><td>Artificial tear ointment to prevent dry eyes while under anesthesia</td></tr><tr><td>Povidine-Iodine Prep Pads</td><td>PDI</td><td>NDC 10819-3883-1</td><td>For surgery; antiseptic</td></tr><tr><td>Alcohol Prep Pads</td><td></td><td></td><td>May be purchased from any standard pharmacy</td></tr><tr><td>Stainless steel surgical scalpel handles (no.3)</td><td>Integra Miltex&amp;nbsp;</td><td>4-7.</td><td>For surgery</td></tr><tr><td>Stainless steel surgical scalpel blades</td><td>Integra Miltex</td><td>4-310 or 4-315</td><td>For surgery; number 10 or 15 scalpel blade</td></tr><tr><td>3% Hydrogen Peroxide</td><td></td><td></td><td>May be purchased from any standard pharmacy</td></tr><tr><td>Micro Clip</td><td>Roboz</td><td>RS-5459</td><td>For surgery, to hold back skin</td></tr><tr><td>00-90 stainless steel machine screw (0.0625&amp;nbsp;in. long)</td><td>Amazon Supply</td><td>B002SG89X4&amp;nbsp;</td><td>For surgery, to wrap ground wire around</td></tr><tr><td>Professional Rotary Tool</td><td>Walnut Hollow</td><td>29637</td><td>Hand drill for surgery, to drill holes in skull</td></tr><tr><td>Inverted Cone Burr</td><td>Roboz</td><td>RS-6282C-34</td><td>Inverted cone burr size 34; for surgery, to drill holes in skull</td></tr><tr><td>Engraving Cutter Drill Bit</td><td>Dremel</td><td>106</td><td>Engraving cutter; 1.6 mm bit; for surgery, to drill holes in skull</td></tr><tr><td>C&amp;amp;B Metabond-Quick! Cement System &quot;B&quot; Quick Base</td><td>Parkell</td><td>S398</td><td>For surgery; adhesive luting cement system; important to prevent headpiece avulsion</td></tr><tr><td>C&amp;amp;B Metabond-Quick! Cement System Clear L-Powder</td><td>Parkell</td><td>S399</td><td>For surgery; adhesive luting cement system; important to prevent headpiece avulsion</td></tr><tr><td>C&amp;amp;B Metabond-Quick! Cement System &quot;C&quot; Universal TBB Catalyst 0.7 ml</td><td>Parkell</td><td>S371</td><td>For surgery; adhesive luting cement system; important to prevent headpiece avulsion</td></tr><tr><td>C&amp;amp;B Metabond-Quick! Cement System Ceramic Mixing Dish with temperature strip</td><td>Parkell</td><td>S387</td><td>For surgery; adhesive luting cement system; important to prevent headpiece avulsion</td></tr><tr><td>Swiss Tweezers, style #5</td><td>World Precision Instruments</td><td>504506</td><td>For surgery</td></tr><tr><td>Puritan Cotton-Tipped Applicators</td><td>VWR International</td><td>10806-005&amp;nbsp;</td><td>For surgery</td></tr><tr><td>Dental Caulk Grip Cement Kit</td><td>Dentsply</td><td>675570</td><td>For surgery; dental cement</td></tr></tbody></table>

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.

8-10周龄雄性C57BL6 / J小鼠接受了关于头部固定圆柱形跑步机设备跟踪眨眼空调。 8小鼠配对CS-美国演讲（条件组）和9只小鼠进行了培训未成对CS和美国的演讲（pseudoconditioned组）进行了培训。 从调节的小鼠的条件性响应的示例肌电图记录示于图3和4。肌电图记录每次试验的进行整流，并用10毫秒的时间常数积分。 图5示出了已整流和综合肌电响应迹线在所有试验中平均每10会话为一个空调（A）和一个pseudoconditioned（B）的小鼠。的条件的响应的演变可以在图5A中可以看出，随着反应越来越大接近美国的发病。这种演变是没见过我n个pseudoconditioned在图5B的反应。 的演变以及定时，因此，良好的学习条件的反应，也可以在图6中看到的， 图6示出的时间的直方图与下列CS发病的响应的峰值。有一个在0之间的图形初始峰值 - CS发病后150毫秒。此初始峰值可以早在会话1.更多的培训课程看出，存在的第二峰的在400-500毫秒的空调动物之间的曲线图的发展，表明更好定时调节应答的发展。 图7A示出了从空调和pseudoconditioned小鼠记录在平均百分之自适应空调的响应。自适应空调响应被认为具有显著活性是存在至少20毫秒BEF美国矿发作（ 即 CS发病前至少4 SD比平均活动存在较大的250毫秒）。在训练课上，空调的小鼠表现出调理范例循序渐进的学习与每次训练更多的参展条件的适应性反应，这样条件的小鼠表现出了比pseudoconditioned小鼠（组，F（1,15）= 20.62，P显著更具适应性的反应条件&lt;.0005;会议，F（1,9）= 9.987，p &lt;0.0001;组*会话，F（1,9）= 5.977，p &lt;0.0001）。注意，pseudoconditioned组通常在试验小于20％表现出伪的CR。 图7B示出了超过十个训练课程都空调和pseudoconditioned小鼠平均条件反应区（整流和综合肌电响应的曲线下的面积）。超过拉斯维加斯重复测量方差分析比较计划ŧ四个交易日（ 即会话7-10，一旦条件组已达到60％，CR的学习标准）显示组的显著主效应，表明CR领域是空调组较pseudoconditioned组学习后大标准（组，F（1,14）= 5.733，p &lt;0.05;会议，F（1,3）= .486，NS;组*会话，F（1,3）= 0.432，NS）。 图7C示出了从空调，并pseudoconditioned小鼠记录在平均百分比的α（惊跳）的反应。阿尔法的反应是在CS发病50毫秒的是至少平均基线活动4SD上述活动。在训练课上，空调，pseudoconditioned通常表现出对试验低于25％的α反应的小鼠，与空调和pseudoconditioned小鼠（组，F（1,15）= 2.502，NS）之间没有显著差异。钍È重复测量ANOVA没有，但是，揭示组和会话的显著相互作用由于pseudoconditioned应答的降低和阿尔法反应在约25％的维修条件的小鼠（组*会话，F（1,9）= 2.074， p &lt;0.05）。会话10期间的α应答的增加可能是由于发达的CR的短潜伏期。 图7D示出了从两组小鼠记录在平均百分之短潜伏期反应。短潜伏期答复如下CS发作，这是4SD平均基线活动上面反映50-70毫秒之间的活动。重复测量方差分析表明，条件的动物表现出了比pseudoconditioned动物更短潜伏期反应了十个培训课程（组，F（1,15）= 5.377，P &lt;0.05;会议，F（1,9）= 3.920，P &lt;.0005;组*会话，F（1,9）= 3.158，p &lt;0.005）。 ŧ他的可能反映在CR的早期发病时间，因为经调节的组与每个训练会话表现出较大的CR。 <img alt="图1" src="/files/ftp_upload/53310/53310fig1.jpg" /> 图1:在跟踪眨眼空调范式在跟踪眨眼空调，无害的条件刺激（CS）是搭配轻度有害的非条件刺激（美国）为了诱导眨眼反应。 CS和美国由自由刺激时空间隙隔开，使得范例海马依赖性23。在这里描述的范例中，一个250毫秒长晶须振动的CS与100配对毫秒长眶周震荡美国。 CS和美国由一个250毫秒的时间长的间隙分开。 <a href="https://www.jove.com/files/ftp_upload/53310/53310fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 图2:该头固定制备 A，头固定设置为与缸，晶须刺激器，和连接器指示的B，晶须刺激与两层弯曲致动器和梳指示眨眼调理。 C，与各个部件的连接器指示:锁定螺钉，7孔带，焊接到插座和环氧树脂保持电线的电线，翅膀从一个TO-220式晶体管D变换，与个人的头盔。份表示:本7孔条，销，并焊接到引脚，包括未涂布的不锈钢地线的电线，和个体休克和EMG不锈钢丝，具有0.2厘米聚酰亚胺涂层的剥离结束即 ，在7孔条每孔为连接器和两个配置头盔，F，A鼠标采用了手术植入头盔，G，A鼠标置于缸内，准备训练。 <a href="https://www.jove.com/files/ftp_upload/53310/53310fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/53310/53310fig3.jpg" /> 图3:例肌电图记录的蓝线划定CS演示文稿（250毫秒长）。红线描出美国介绍，这表明来自电击（100毫秒）的工件。 CS和美国是由250毫秒无刺激间隔隔开。在这个无刺激间隔较高幅度肌电活动（相对于基线），它描绘了一个CR。 <a href="https://www.jove.com/files/ftp_upload/53310/53310fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。&lt;</a>/ A&gt; <img alt="图4" src="/files/ftp_upload/53310/53310fig4.jpg" /> 图4:实施例肌电图和相应的光的红外反射记录肌电图记录是正如在检测条件响应发病（蓝线），等待时间来响应峰（红线）的光的红外反射传感器，和无条件眨眼反应发作（准确粉红色线）。 <a href="https://www.jove.com/files/ftp_upload/53310/53310fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图5" src="/files/ftp_upload/53310/53310fig5.jpg" /> 图5:场均眨眼痕迹场均整流，一个鼠标调节（A）和一个pseudoconditioned鼠标（B）的眨眼反应的综合痕迹。每个跟踪代表平均鼠标超过一个会话的所有试验的反应。蓝色部分代表晶须振动CS演示过程中的响应。红色部分表示无刺激的跟踪间隔期间的响应。在A，冲击美国引起的神器，它是跟踪时间间隔后深灰色区域时在座。B显示的晶须振动CS只有响应。 CS单独介绍和美国单独试验是在pseudoconditioning pseudorandomized。 <a href="https://www.jove.com/files/ftp_upload/53310/53310fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图6" src="/files/ftp_upload/53310/53310fig6.jpg" /> 图6:时间到响应峰值响应高峰时段直方图所有的直方图空调（N = 8;蓝色条）和8 pseudoconditioned（红色条）动物的所有TR在所有会话IALS（从一个pseudoconditioned鼠标数据被排除，以允许反应基团之间的数的直接比较）。紫条指明的空调，并pseudoconditioned动物重叠。响应高峰时间的计算公式为CS和美国之间发生的整顿和整合肌电图记录的最大峰值的时间。 <a href="https://www.jove.com/files/ftp_upload/53310/53310fig6large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图7" src="/files/ftp_upload/53310/53310fig7.jpg" /> 图7:调节的动物的响应曲线（N = 8;蓝色曲线）和pseudoconditioned动物（N = 9;红色曲线）。 A，百分比场均适应性条件反射（±SEM）的条件和pseudoconditioned动物超过10培训班。B，场均面积为CR条件（±SEM）和pseudoconditioned动物10多个培训班。C，百分比平均每阿尔法响应（±SEM）的条件和pseudoconditioned动物超过10培训班。对条件D，百分比场均短潜伏期响应（±SEM）和pseudoconditioned过10届。 <a href="https://www.jove.com/files/ftp_upload/53310/53310fig7large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Whisker-signaled Eyeblink Classical Conditioning in Head-fixed Mice at JoVE.com

Whisker-signaled Eyeblink Classical Conditioning in Head-fixed Mice

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 ...

whisker-signaled-eyeblink-classical-conditioning-in-head-fixed-mice

Nanyang Technological University

Research

JoVE Journal

Behavior

12.9K 次观看.  Northwestern University. 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.

视频: Whisker-signaled Eyeblink Classical Conditioning in Head-fixed Mice

Flat-floored Air-lifted Platform: A New Method for Combining Behavior with Microscopy or Electrophysiology on Awake Freely Moving Rodents

It is widely acknowledged that the use of general anesthetics can undermine the relevance of electrophysiological or microscopical data obtained from a living animal&#8217;s brain. Moreover, the lengthy recovery from anesthesia limits the frequency of repeated recording/imaging episodes in longitudinal studies. Hence, new methods that would allow stable recordings from non-anesthetized behaving mice are expected to advance the fields of cellular and cognitive neurosciences. Existing solutions range from mere physical restraint to more sophisticated approaches, such as linear and spherical treadmills used in combination with computer-generated virtual reality. Here, a novel method is described where a head-fixed mouse can move around an air-lifted mobile homecage and explore its environment under stress-free conditions. This method allows researchers to perform behavioral tests (e.g., learning, habituation or novel object recognition) simultaneously with two-photon microscopic imaging and/or patch-clamp recordings, all combined in a single experiment. This video-article describes the use of the awake animal head fixation device (mobile homecage), demonstrates the procedures of animal habituation, and exemplifies a number of possible applications of the method.

This method creates a tangible, familiar environment for the mouse to navigate and explore during microscopic imaging or single-cell electrophysiological recordings, which require firm fixation of the animal&#8217;s head.

平地板的空气解禁平台:一种新方法与显微镜或电对清醒结合行为自由移动鼠害

It is widely acknowledged that the use of general anesthetics can undermine the relevance of electrophysiological or microscopical data obtained from a ...

科研

行为学

Introducing Clicker Training as a Cognitive Enrichment for Laboratory Mice

Establishing new refinement strategies in laboratory animal science is a central goal in fulfilling the requirements of Directive 2010/63/EU. Previous research determined a profound impact of gentle handling protocols on the well-being of laboratory mice. By introducing clicker training to the keeping of mice, not only do we promote the amicable treatment of mice, but we also enable them to experience cognitive enrichment. Clicker training is a form of positive reinforcement training using a conditioned secondary reinforcer, the "click" sound of a clicker, which serves as a time bridge between the strengthened behavior and an upcoming reward. The effective implementation of the clicker training protocol with a cohort of 12 BALB/c inbred mice of each sex proved to be uncomplicated. The mice learned rather quickly when challenged with tasks of the clicker training protocol, and almost all trained mice overcame the challenges they were given (100% of female mice and 83% of male mice). This study has identified that clicker training for mice strongly correlates with reduced fear in the mice during human-mice interactions, as shown by reduced anxiety-related behaviors (e.g., defecation, vocalization, and urination) and fewer depression-like behaviors (e.g., floating). By developing a reliable protocol that can be easily integrated into the daily routine of the keeping of laboratory mice, the lifetime experience of welfare in the mice can be improved substantially.

The development of new refinement strategies for laboratory mice is a challenging task that contributes towards fulfilling the 3R principle. This protocol introduces clicker training as a cognitive enrichment program for laboratory mice.

引进响片训练为实验室老鼠认知富集

Establishing new refinement strategies in laboratory animal science is a central goal in fulfilling the requirements of Directive 2010/63/EU. Previous ...

Classical Short-Delay Eyeblink Conditioning in One-Year-Old Children

Classical eyeblink conditioning (EBC) refers to the learned association between a conditioned stimulus (an auditory tone) and an unconditioned stimulus (a puff of air to the cornea). Eyeblink conditioning is often used experimentally to detect abnormalities in cerebellar-dependent learning and memory that underlies this type of associative learning. While experiments in adults and older children are relatively simple to administer using commercial equipment, eyeblink conditioning in infants is more challenging due to their poor compliance, which makes correct positioning of the equipment difficult. To achieve conditioning in one-year-old infants, a custom-made or an adapted commercial system can be used to deliver the air puff to the infant's cornea. The main challenge lies in successfully detecting and classifying the behavioral responses. We report that automated blink detection methods are unreliable in this population, and that conditioning experiments should be analyzed using frame-by-frame analysis of supplementary video camera recordings. This method can be applied to study developmental changes in eyeblink conditioning and to examine whether this paradigm can detect children with neurological disorders.

This protocol describes an eyeblink conditioning paradigm appropriate for experiments with one-year-old infants. Commercial or custom-made equipment can be used to deliver the stimuli, and data collection and analysis should be performed on the video recordings.

经典短时滞上只是一眨眼在一岁儿童中的调节作用

Classical eyeblink conditioning (EBC) refers to the learned association between a conditioned stimulus (an auditory tone) and an unconditioned stimulus (a ...

A System for Tracking the Dynamics of Social Preference Behavior in Small Rodents

Exploring the neurobiological mechanisms of social behavior requires behavioral tests that can be applied to animal models in an unbiased and observer-independent manner. Since the beginning of the millennium, the three-chamber test has been widely used as a standard paradigm to evaluate sociability (social preference) and social novelty preference in small rodents. However, this test suffers from multiple limitations, including its dependence on spatial navigation and negligence of behavioral dynamics. Presented and validated here is a novel experimental system that offers an alternative to the three-chamber test, while also solving some of its caveats. The system requires a simple and affordable experimental apparatus and publicly available open-source analysis system, which automatically measures and analyzes multiple behavioral parameters at individual and population levels. It allows detailed analysis of the behavioral dynamics of small rodents during any social discrimination test. We demonstrate the efficiency of the system in analyzing the dynamics of social behavior during the social preference and social novelty preference tests as performed by adult male mice and rats. Moreover, we validate the ability of the system to reveal modified dynamics of social behavior in rodents following manipulations such as whisker trimming. Thus, the system allows for rigorous investigation of social behavior and dynamics in small rodent models and supports more accurate comparisons between strains, conditions, and treatments.

Described here is a novel automated experimental system that offers an alternative to the three-chamber test and also solves several caveats. This system supplies multiple behavioral parameters that enable rigorous analysis of small rodent behavioral dynamics during the social preference and social novelty preference tests.

跟踪小啮齿动物社会偏好行为动态的系统

Exploring the neurobiological mechanisms of social behavior requires behavioral tests that can be applied to animal models in an unbiased and ...

Investigating Migraine-Like Behavior Using Light Aversion in Mice

Migraine is a complex neurological disorder characterized by headache and sensory abnormalities, such as hypersensitivity to light, observed as photophobia. Whilst it is impossible to confirm that a mouse is experiencing migraine, light aversion can be used as a behavioral surrogate for the migraine symptom of photophobia. To test for light aversion, we utilize the light/dark assay to measure the time mice freely choose to spend in either a light or dark environment. The assay has been refined by introducing two critical modifications: pre-exposures to the chamber prior to running the test procedure and adjustable chamber lighting, permitting the use of a range of light intensities from 55 lux to 27,000 lux. Because the choice to spend more time in the dark is also indicative of anxiety, we also utilize a light-independent anxiety test, the open field assay, to distinguish anxiety from light-aversive behavior. Here, we describe a modified test paradigm for the light/dark and open field assays. The application of these assays is described for intraperitoneal injection of calcitonin gene-related peptide (CGRP) in two mouse strains and for optogenetic brain stimulation studies.

Rodents are not able to report migraine symptoms. Here, we describe a manageable test paradigm (light/dark and open field assays) to measure light aversion, one of the most common and bothersome symptoms in patients with migraines.

使用轻度厌恶研究小鼠偏头痛样行为

Migraine is a complex neurological disorder characterized by headache and sensory abnormalities, such as hypersensitivity to light, observed as ...

Juxtacellular Monitoring and Localization of Single Neurons within Sub-cortical Brain Structures of Alert, Head-restrained Rats

There are a variety of techniques to monitor extracellular activity of single neuronal units. However, monitoring this activity from deep brain structures in behaving animals remains a technical challenge, especially if the structures must be targeted stereotaxically. This protocol describes convenient surgical and electrophysiological techniques that maintain the animal&#8217;s head in the stereotaxic plane and unambiguously isolate the spiking activity of single neurons. The protocol combines head restraint of alert rodents, juxtacellular monitoring with micropipette electrodes, and iontophoretic dye injection to identify the neuron location in post-hoc histology. While each of these techniques is in itself well-established, the protocol focuses on the specifics of their combined use in a single experiment. These neurophysiological and neuroanatomical techniques are combined with behavioral monitoring. In the present example, the combined techniques are used to determine how self-generated vibrissa movements are encoded in the activity of neurons within the somatosensory thalamus. More generally, it is straightforward to adapt this protocol to monitor neuronal activity in conjunction with a variety of behavioral tasks in rats, mice, and other animals. Critically, the combination of these methods allows the experimenter to directly relate anatomically-identified neurophysiological signals to behavior.

This protocol describes the design and surgical implantation of a head-restraining mechanism to monitor neuronal activity in sub-cortical brain structures in alert rats. It delineates procedures to isolate single neurons in the juxtacellular configuration and to efficiently identify their anatomical locations.

警报，头抑制大鼠子皮层的大脑结构中Juxtacellular监测和本地化单神经元

There are a variety of techniques to monitor extracellular activity of single neuronal units. However, monitoring this activity from deep brain structures ...

神经科学

Behavioral Training Procedures for Head-fixed Virtual Reality in Mice

Virtual reality (VR) combined with head-fixation is increasingly being utilized in behavioral neuroscience studies as it allows complex behavioral assays to be performed in head-fixed mice. This enables precise behavioral recordings while incorporating various neurophysiological techniques that require head-fixation to minimize movement-related signal noise during neural recordings. However, despite the growing use of VR, there is little published data on the detailed methodology of how to implement it. In this study, a training protocol is developed whereby male and female C57B16/J mice are trained to run down a virtual linear corridor, the length of which is increased from 1-3 m over multiple training sessions. Building upon this foundation, this study investigated the feasibility of mice performing complex behaviors within VR using a Y-maze paradigm. The task required navigating to the arm with black walls from the choice point in the Y-maze. After reaching a criterion of two consecutive days equal to or greater than 70% correct, the mice progressed to increasingly difficult sensory discrimination. The findings provide important details on the methodologies useful for the successful training of mice in VR and demonstrate that mice exhibit learning capabilities in navigating the Y-maze. The methodology presented not only offers insights into training duration in VR-based assays but also underscores the potential for probing intricate behaviors in mice, opening avenues for more comprehensive neuroscience investigations.

The article describes the experimental procedures for the commonly used linear track virtual reality (VR) paradigm in mice as well as determining the feasibility of running complex VR tasks by testing a Y-shaped signal discrimination task.

小鼠头部固定虚拟现实的行为训练程序

Virtual reality (VR) combined with head-fixation is increasingly being utilized in behavioral neuroscience studies as it allows complex behavioral assays ...

<meta charset="utf8"></head><body>使用 DeepLabCut 实时小鼠斜视检测偏头痛追踪

An Automated Squint Method for Time-syncing Behavior and Brain Dynamics in Mouse Pain Studies

Spontaneous pain has been challenging to track in real time and quantify in a way that prevents human bias. This is especially true for metrics of head pain, as in disorders such as migraine. Eye squint has emerged as a continuous variable metric that can be measured over time and is effective for predicting pain states in such assays. This paper provides a protocol for the use of DeepLabCut (DLC) to automate and quantify eye squint (Euclidean distance between eyelids) in restrained mice with freely rotating head motions. This protocol enables unbiased quantification of eye squint to be paired with and compared directly against mechanistic measures such as neurophysiology. We provide an assessment of AI training parameters necessary for achieving success as defined by discriminating squint and non-squint periods. We demonstrate an ability to reliably track and differentiate squint in a CGRP-induced migraine-like phenotype at a sub second resolution.

<meta charset="utf8"></head><body>作者聚焦：破译复杂大脑活动和疾病背后的电网

This protocol provides a method for tracking automated eye squint in rodents over time in a manner compatible with time-locking to neurophysiological measures. This protocol is expected to be useful to researchers studying mechanisms of pain disorders such as migraine.

一种用于小鼠疼痛研究中时间同步行为和大脑动力学的自动斜视方法

Spontaneous pain has been challenging to track in real time and quantify in a way that prevents human bias. This is especially true for metrics of head ...

在头固定小鼠胡须信号眨眼经典条件反射

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