Name: 视频: Video-oculography in Mice
Uploaded: 2012-07-19T12:00:00.000+00:00
Duration: 9 min 43 s
Description: 23.6K 次观看. Erasmus MC, Rotterdam, The Netherlands. 的视频oculography是一个非常量化的方法来研究眼球运动表现以及运动学习。在这里，我们描述如何衡量小鼠视频oculography。对正常应用这项技术，药物治疗或转基因小鼠是一个强有力的研究工具，探索潜在的生理运动行为。

我们诚挚地感谢荷兰健康研究与发展组织（牡丹江，CDZ），荷兰科学研究组织（CDZ），NeuroBasic（CDZ），（CDZ）贝娅特丽克丝全宗，"SENSOPAC（CDZ），C7（CDZ）和Prinses CEREBNET（CDZ）欧洲共同体的财政支持方案。

1。准备与杜赫动物实验伦理委员会根据以下实验进行。 <ol><li>准备视频-oculography的小鼠。为了测量眼球运动鼠标，鼠标的头部，需要被固定。因此，一个基座建设鼠标头骨（ 图1）。 <ol><li>麻醉由异氟醚的混合物（异氟醚1-1.5％，法国罗地亚Organique精细有限公司）鼠标和氧气在毒气室。过多的气体被清除。通过鼻锥维持麻醉。通过脚趾掐确认的麻醉深度。 </li><li>肛门热敏使用和加热垫（金控，Bowdoinham，ME）的体温保持在37°C。 </li><li>眼膏（duratears，爱尔康，比利时）覆盖保护他们的眼睛。刮胡子背颅毛皮的，清理手术区旋转擦洗和betadine或洗必泰溶液。 </li><li>使中间线切口，暴露头骨的颅背表面。使表面的清洁和干燥。 </li><li>应用磷酸（磷酸凝胶蚀刻37.5％;克尔，CA）的下降，从囟门的lambda头骨的颅背表面上。 15秒后取出腐蚀剂和面颅再次盐水和干燥清洁。 </li><li>申请下降了OptiBond总理（科尔，CA）和空气干燥30秒这个表面蚀刻颅顶部。 </li><li>放置的顶级的OptiBond总理胶粘剂OptiBond（科尔，CA）的下降，治愈1分钟（千里马480可见光固化装置;亨利施恩，美国）。 </li><li>覆盖一层薄薄的魅力复合贺利氏古莎（德国）的粘接层。嵌入两个连接螺母（直径3毫米）的复合材料。与光固化复合事后。必要时，申请额外的复合层与光固化。 </li><li>广告部长丁丙诺啡（0.015毫克/公斤，SC）手术后的镇痛。约5分钟内的动物应该是它的脚。允许鼠标在室温中恢复，手术后至少3天在家里笼。 </li></ol></li><li>对小鼠的视频oculography设置（ 图2）。 <ol><li>鼠标放置在限位和修复的拦阻他的头两个螺丝（ 图1）。鼠标不需要麻醉此过程。限制时间不应超过1小时/天。 </li><li>安装鼠标的头部和身体的XY平台上的拦阻，而这又是后安装的转盘（直径:60厘米）。使用XY平台，可以放置鼠标头以上的转盘中心。移动鼠标可以在俯仰，偏航和辊轴。鼠标的头部被放置在正确的俯仰，偏航和横滚角对准眼睛，用眼，由ISCAN或连结产生的视觉形象米。另外，基座建设，可以放在鼠标的头在一个立体的框架11。 </li><li>转盘连接到交流伺服控制电机（谐波传动公司，荷兰）和转盘的位置进行监测转盘轴连接一个电位（BOURNS公司，加利福尼亚）。 </li><li>圆柱形（直径:63厘米高度:35厘米），与周围的屏幕随机点模式（每个元素2°），涵盖了转盘，这个鼓也配备交流伺服控制电机（谐波传动公司，荷兰） 。监测的圆柱形屏幕上的位置由一个电位连接轴和屏幕可以点亮一盏卤素灯（20瓦）（BOURNS公司，加利福尼亚）。周围的屏幕和转盘独立驱动。 </li><li>转盘和屏幕周围的运动控制由计算机连接到一个I / O接口（海关有限，剑桥，英国）。钽BLE和周围的屏幕上的位置信号进行过滤（截止频率:20赫兹），I / O接口的数字化和存储在这台电脑上。 </li><li>三个红外线发射器照射老鼠的眼睛（600兆瓦，分散角度:7°，峰值波长:880纳米，RS元件，荷兰）。两个红外线发射器是固定在转盘上，第三个发射器连接到相机。这第三个发射器产生一个参考角膜反射（CR），这是在校准过程中和在眼球运动记录。 </li><li>配备变焦镜头（NAVITAR公司变焦6000，纽约）一个红外CCD相机连接到转盘和转盘的中心集中在鼠标头。相机可以完全超过20°解锁并可以在校准过程中的转盘轴yawed。 </li><li>视频信号处理系统的ETL-200，ISCAN，伯灵顿，马眼跟踪。该的ISCAN系统使用算法rithm追踪瞳孔和参考CR中心。该系统可以追踪在水平和垂直方向的瞳孔和参考CR在120赫兹的采样率。 </li><li>参考CR的位置，瞳孔位置和瞳孔大小的信号通过I / O接口的数字化，并存储在相同的文件表和周围的屏幕上的位置信号。视频瞳孔跟踪系统引起的眼球运动信号约27毫秒的延迟。 </li></ol></li></ol> 2。使用视频瞳孔跟踪校准和测量眼球运动眼球跟踪系统捕获的瞳孔作为平移运动的运动。追踪瞳孔的平移运动包含平移的组成部分，由于旋转中心的眼睛和眼睛的角膜曲率中心（即）的解剖中心，由于眼球的旋转角度和旋转组件之间的轴向差异。通过减不受欢迎的翻译部分参考CR ING从瞳孔运动/位置，被淘汰的信号，致使眼球的旋转，这只是由于在平移运动。虽然他们往往非常小，减法也消除了头部和相机之间的翻译。 8,12以下的标定方法，残余的孤立的平移运动转换成角旋转眼球。该校准之前进行任何眼动实验。 <ol><li>调整鼠标的头部位置的摄像头，视频图像的瞳孔位于显示器的中间，位于以上，最好是直接的眼睛瞳孔的垂直中线的参考CR表示，在这样一种方式。最小化的参考CR运动，由于角度的摄像机旋转，可放置在摄像头/表轴角膜曲率中心完成。/ LI&gt; <li>几次旋转的摄像头+ / - 10°垂直轴周围的转盘（即20度的峰峰值）。使用追踪瞳孔（P）和记录在摄像机的旋转的极端立场的参考CR的位置来计算的瞳孔的旋转半径（RP; RP =Δ/罪（20°），其中Δ=（CR -P）参见图3A）。 </li><li>由于RP值瞳孔的大小取决于事实，瞳孔大小校正需要实现12（ 图3B）。重复加强2.2多次在各种光照条件下（即操纵瞳孔的大小， 图3C），以确定瞳孔的大小- RP的关系，并组成一个RP校正曲线（ 图3D）。 RP值还取决于垂直眼位。实验时将导致垂直眼球运动，然后垂直眼睛位置的校准校正是非常可取的13。 </li><li>通过测量参考CR的位置，P位置和瞳孔大小的眼睛（五）确定的角位置。参考CR位置减去学生产生平移免费瞳孔位置的位置。通过测量瞳孔大小的RP值可以提取RP校正曲线和E可以通过使用下列公式计算Ë= arcsin（Δ1）/ RP}（ 图4A，Δ1=（2 - P 1）和P 1和P 2修正的参考CR减法）。 </li><li>一个转盘和/或周围的屏幕旋转的大型剧目，现在可以用来刺激动眼神经系统。为了执行老鼠的眼睛在黑暗中视频oculography，必须用缩瞳药物预处理限制瞳孔扩张，并允许在这种情况下，瞳孔跟踪。在我们的实验中，我们用毛果芸香碱（4％，肖实验室，法国），以限制瞳孔扩张黑暗中。 </li></ol> 3。数据分析<ol><li>眼睛的位置，表的位置和周围的屏幕位置都转换成角的位置（ 见图4B和公式2.4）。眼科信号延迟27毫秒诱导其纠正，由瞳孔跟踪系统的图像处理。 </li><li>眼表及周边屏幕的角位置是有区别的，巴特沃斯低通使用截止频率为20赫兹的切割过滤器过滤。 </li><li>扫视眼使用检测阈值的40°/ s的速度信号被删除。数据被删除过路的检测阈值后，开始前，高达80毫秒20毫秒。 </li><li>在每一个人的踪迹（ 图4C）周期表，周围的屏幕和眼睛的速度信号的平均值。 </li><li>平均信号都配有相应的功能。在一般情况下，使用正弦速度刺激和平均周期配备窦或cosinus功能（ 图4C）。然后，增益可眼速度的刺激速度的比值计算，可作为眼睛的速度和刺激的速度之间的差异（度）计算，而阶段。 </li></ol> 4。代表结果 ，视频oculography可以用来探讨各种的动眼神经表演形式（即视动反射:OKR;前庭眼反射:VOR的视觉增强前庭眼反射:VVOR）以及电机学（VOR的适应; OKR适应）。 OKR补偿使用视觉反馈的低频干扰。 OKR旋转的周围照屏幕（ 电影1）可诱发。旋转超过0.2 -1.0幅度为1.6°赫兹的频率范围内的周边屏幕显示的视动系统在低频率范围内THA是一种更有效的补偿机制如何n在高频率范围内（ 图5A）。 VOR的补偿高频头使用信号从前庭器官的运动。在黑暗中（ 电影2）动物（即转盘）旋转的VOR可诱发。超过幅度为1.6°，0.2 -1.0赫兹的频率范围旋转的转盘，前庭眼系统演示了如何更有效的补偿眼球运动产生的高频率范围比在低频率范围（ 图5A） 。在演唱会视动和前庭眼系统的行为时，图像可以稳定在视网膜上的头部运动的广泛。超过幅度为1.6°，0.2 -1.0赫兹的频率范围旋转的转盘，而周围的屏幕照（ 电影3）眼如何产生"高"在整个频率范围内增益补偿运动（ 图5A ）。所有这些增益和pH值ASE值是为小鼠典型，虽然性别14和应变15,16,17差异。 在转盘和周围的屏幕的独立控制，使我们面对与老鼠之间的视觉和前庭信息不匹配。长期不匹配的视觉和前庭信息和统一的接触后，鼠标的VOR会改变，以弥补改变视觉输入（VOR的适应; 电影4）。旋转转盘阶段（即180°）与周边屏幕（1赫兹，1.6°），增加的VOR增益（ 图5B）。中的VOR增益的最大变化，当使用一个审判的学习范例，往往达到30分钟后。 <img alt="图1" src="/files/ftp_upload/3971/3971fig1.jpg" /> 图1。鼠标的头部和身体限位示意图绘制。鼠标的身体受到抑制使用直径35毫米的塑料圆筒形管。鼠标的头部被铁棍连接鼠标的底座，用两个螺钉固定。铁条，使30度角，以头鼠标定位在行走过程中的正常间距。 *，包含两个螺母座的顶视图。 <img alt="图2" src="/files/ftp_upload/3971/3971fig2.jpg" /> 图2。示意图的鼠标视频oculography的设置。 <img alt="图3" src="/files/ftp_upload/3971/3971fig3.jpg" /> 图3。视频瞳孔跟踪系统的校准。 A）该相机+ / - 10°（即20度，峰峰值）围绕垂直轴转盘旋转几次。跟踪的瞳孔（P）和参考角膜反射（CR）记录在摄像机的旋转的极端立场是用来计算的瞳孔半径旋转（RP）。乙）瞳孔直径的半径是依赖于瞳孔的大小。 c）示例显示在校准过程中的瞳孔大小瞳孔位置的影响（无论是在像素（px））。四）Rp和瞳孔直径之间的关系，在一个单一的鼠标。完成了13个不同的瞳孔直径，通过改变周围的光线强度。 <img alt="图4" src="/files/ftp_upload/3971/3971fig4.jpg" /> 图4。使用视频瞳孔跟踪测量和分析眼球运动。一）角瞳孔位置计算半径（RP）的瞳孔和瞳孔的位置（P，为华润位置纠正）。乙）代偿眼球运动为例诱导刺激前庭和视觉系统（视觉增强的VOR）。转盘旋转正弦幅度为1.6°，0.6赫兹，而周围的屏幕上是良好的照明。 c）分析记录在B所示）。图表显示平均速度跟踪转盘（蓝色）和学生（红色）。这些平均痕迹装有一个正弦函数（黑色）。 <img alt="图5" src="/files/ftp_upload/3971/3971fig5.jpg" /> 图5。一个C57BL6鼠标测量动眼神经系统的性能和学习。周围的屏幕旋转（视动反射:OKR，顶部面板），通过旋转鼠标在黑暗中（前庭眼反射:VOR的，中板），并在光通过旋转鼠标（视）眼球运动所产生的增强前庭眼反射:VVOR，底部面板）频率范围从0.2到1.0赫兹的幅度为1.6°。从眼睛的速度和刺激的速度（右面板）之间的相位差计算眼速度的刺激速度的比值（左侧面板）和相位的反射，反射的增益计算。二）汽车学习完成<html>自适应增加的VOR使用了一个阶段的训练范例。鼠标到visuovestibular培训范式鼠标旋转是与周边屏幕，四十分钟（均在1.0赫兹，1.6°旋转）旋转（180°）相。每隔10分钟测试的VOR（1.0赫兹，1.6°）。在这种鼠标阶段的训练了增加的VOR增益。 电影1。动画显示的范例，诱导小鼠OKR <a href="http://www.jove.com/files/ftp_upload/3971/3971movie1.mov" target="_blank">点击这里观看电影</a> 。 电影2动画显示诱导小鼠的VOR范式。 <a href="http://www.jove.com/files/ftp_upload/3971/3971movie2.mov" target="_blank">点击这里观看电影</a> 。 电影3。动画显示的范例诱导小鼠VVOR的。.com/files/ftp_upload/3971/3971movie3.mov"目标="_blank"&gt;点击这里观看电影。 电影4。动画显示visuovestibular出诱导小鼠（增加）的VOR适应阶段的训练范例。 <a href="http://www.jove.com/files/ftp_upload/3971/3971movie4.mov" target="_blank">点击这里观看电影</a> 。

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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eye+movements+of+the+murine+P/Q+calcium+channel+mutant+tottering,+and+the+impact+of+aging.">Eye movements of the murine P/Q calcium channel mutant tottering, and the impact of aging.</a> J. Neurophysiol. 95, 1588 (2006).</li><li>Andreescu, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estradiol+improves+cerebellar+memory+formation+by+activating+estrogen+receptor+beta.">Estradiol improves cerebellar memory formation by activating estrogen receptor beta.</a> Journal of Neuroscience. 27, 10832 (2007).</li><li>Katoh, A., Kitazawa, H., Itohara, S., Nagao, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+characteristics+and+adaptability+of+mouse+vestibulo-ocular+and+optokinetic+response+eye+movements+and+the+role+of+the+flocculo-olivary+system+revealed+by+chemical+lesions.">Dynamic characteristics and adaptability of mouse vestibulo-ocular and optokinetic response eye movements and the role of the flocculo-olivary system revealed by chemical lesions.</a> Proc. Natl. Acad. Sci. U.S.A. 95, 7705 (1998).</li><li>Stahl, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Using+eye+movements+to+assess+brain+function+in+mice.">Using eye movements to assess brain function in mice.</a> Vision Res. 44, 3401 (2004).</li><li>Koekkoek, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gain+adaptation+and+phase+dynamics+of+compensatory+eye+movements+in+mice.">Gain adaptation and phase dynamics of compensatory eye movements in mice.</a> Genes Funct. 1, 175 (1997).</li><li>Faulstich, B. M., Onori, K. A., du Lac, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+plasticity+and+development+of+mouse+optokinetic+and+vestibulo-ocular+reflexes+suggests+differential+gain+control+mechanisms.">Comparison of plasticity and development of mouse optokinetic and vestibulo-ocular reflexes suggests differential gain control mechanisms.</a> Vision Res. 44, 3419 (2004).</li><li>Koekkoek, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gain+adaptation+and+phase+dynamics+of+compensatory+eye+movements+in+mice.">Gain adaptation and phase dynamics of compensatory eye movements in mice.</a> Genes Funct. 1, 175 (1997).</li></ol>

没有利益冲突的声明。

为了获得高品质的视频眼球运动记录小鼠几项要求是必要的。校准过程必须在上述的标准化问题。例如偏离中心的校准时，瞳孔不上的参考CR垂直中线定位在校准过程中，会导致的RP被低估，因此高估的眼球运动。此外，我们建议整合在校准过程12瞳孔大小的校正方法，因为试验表明一个非常稳定的瞳孔大小是非常罕见的。即使是在审讯过程中的小应激已经可以大幅改变瞳孔直径。 眼动实验设计时，必须考虑下列因素或控制，因为它们是已知的影响眼球运动反应:14 13,18年龄，性别和应变15,1619。此外，实验动物应该有色素的虹膜是不可能的，当瞳孔和虹膜之间的对比度太低，像在BALB / c小鼠，因为瞳孔检测和跟踪。极度紧张或焦虑的动物需要进行培训，实验前，习惯于实​​验设置和约束条件。这种动物处理过程的结果在更短的封闭或半封闭的眼睛，并防止在实验过程中产生眼流体，从而实现更好的瞳孔追踪。 最后，获取和分析数据，需要两到三个小时，每头牲畜。因此，眼球运动记录可能会保持一个特定的程序，适用于选择小鼠，不适合作为一种高通量筛选试验。

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td>异氟醚</td><td>罗地亚Organique精细有限公司</td><td></td></tr><tr><td>加热垫</td><td>金控</td><td> 40-90-8 </td></tr><tr><td> duratears </td><td>爱尔康</td><td></td></tr><tr><td>磷酸凝胶</td><td>克尔</td><td> 31297 </td></tr><tr><td> optibond总理</td><td>克尔</td><td> 35369 </td></tr><tr><td> optibond胶粘剂</td><td>克尔</td><td> 35369 </td></tr><tr><td>魅力复合</td><td>贺利氏古莎</td><td></td></tr><tr><td>千里马480光固化机</td><td>亨利施恩</td><td></td></tr><tr><td>交流伺服控制电机</td><td>谐波传动公司</td><td></td></tr><tr><td>共青团indric屏幕</td><td></td><td></td></tr><tr><td>卤素灯（20瓦） </td><td>欧时电子元件</td><td></td></tr><tr><td>电位器（精密） </td><td> BOURNS INC。 </td><td> 6574 </td></tr><tr><td> 1401电源（I / O接口） </td><td>海关有限</td><td></td></tr><tr><td>电脑</td><td>戴尔</td><td></td></tr><tr><td>红外emmitters </td><td>欧时电子元件</td><td> 195-451 </td></tr><tr><td> ETL - 200 </td><td> ISCAN </td><td></td></tr><tr><td>变焦镜头（变焦6000） </td><td> Navitar的公司。 </td><td></td></tr><tr><td> pilocarpinenitrate（minims） </td><td> LABORATOIRE肖</td><td></td></tr></tbody></table>

video-oculography in mice

为了跟踪一个对象或稳定在视网膜上的图像在运动，眼球运动是非常重要的。没有凹的动物，如鼠标，有能力是有限的目标锁定到他们的眼睛。相反，这些目标导向的眼球运动，代偿眼眼球运动很容易，引起在1,2,3,4 afoveate的动物。补偿眼球运动所产生的处理前庭和视动到一个命令信号的信息，这将推动眼部肌肉。前庭和视动性信息的处理，可以单独和共同研究，使赤字在动眼神经系统的规范。动眼神经系统可以唤起视动反射（OKR），前庭眼反射（VOR）或视觉增强前庭眼反射（VVOR）测试。 OKR是一种反射运动，为"全领域"在视网膜上的图像运动补偿，而VOR是一个反射眼Movement补偿头部运动。 VVOR是反射眼球运动，使用两个前庭以及视动性信息，以作出适当的赔偿。小脑监控，并能够适应这些补偿性眼球运动。因此，oculography是一个非常强大的工具，以探讨在正常脑行为的关系，以及病理条件下（前庭，眼和/或小脑的起源FE）。 测试动眼神经系统，作为一种行为模式，有趣的是以下几个原因。首先，动眼神经系统是一个理解的神经系统。第二，动眼神经系统是相对简单的6球插座架构（"单联"）和额外的眼部肌肉7三对可能眼球运动的数额由有限。第三，行为输出和感官输入可以很容易地进行测量，这使得这对定量一个非常方便的系统分析8。许多行为测试缺乏这定量电源的高层次。最后，动眼神经系统的性能以及可塑性可以进行测试，使研究学习和记忆过程9。 时下广泛使用的转基因小鼠，它们形成了一个在探索10个不同层次的脑功能的重要来源。此外，他们还可以用来作为模型来模拟人类疾病。应用的正常oculography，药物治疗或转基因小鼠是一个强有力的研究工具，以发掘潜在的生理正常和病理条件下的运动行为。在这里，我们描述如何衡量在小鼠8视频oculography。

Watch this Scientific Journal Video about Video-oculography in Mice at JoVE.com

Video-oculography in Mice

的视频oculography是一个非常量化的方法来研究眼球运动表现以及运动学习。在这里，我们描述如何衡量小鼠视频oculography。对正常应用这项技术，药物治疗或转基因小鼠是一个强有力的研究工具，探索潜在的生理运动行为。

视频oculography小鼠

Eye movements are very important in order to track an object or to stabilize an image on the retina during movement. Animals without a fovea, such as the ...

Research

JoVE Journal

Neuroscience

23.6K 次观看.  Erasmus MC, Rotterdam, The Netherlands. 的视频oculography是一个非常量化的方法来研究眼球运动表现以及运动学习。在这里，我们描述如何衡量小鼠视频oculography。对正常应用这项技术，药物治疗或转基因小鼠是一个强有力的研究工具，探索潜在的生理运动行为。

视频: Video-oculography in Mice

Measuring Motor Coordination in Mice

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such as emotionality or cognition can be assessed, however, it is vital to determine whether the motor capabilities of e.g. a mutant or lesioned mouse allow such an assessment. Performance on a maze task requiring strength and coordination, such as the Morris water maze, might well be impaired in a mouse by motor, rather than cognitive, impairments, so it is essential to selectively dissect the latter from the former. For example, sensorimotor impairments caused by NMDA antagonists have been shown to impair water maze performance2. 
Motor coordination has traditionally been assessed in mice and rats by the rotarod test, in which the animal is placed on a horizontal rod that rotates about its long axis; the animal must walk forwards to remain upright and not fall off. Both set speed and accelerating versions of the rotarod are available.
The other three tests described in this article (horizontal bar, static rods and parallel bars) all measure coordination on static apparatus. The horizontal bar also requires strength for adequate performance, particularly of the forelimbs as the mouse initially grips the bar just with the front paws. 
Adult rats do not perform well on tests such as the static rods and parallel bars (personal observations); they appear less well coordinated than mice. I have only tested male rats, however, and male mice seem generally less well coordinated than females. Mice appear to have a higher strength:weight ratio than rats; the Latin name, Mus musculus, seems entirely appropriate. The rotarod, the variations of the foot fault test12 or the Catwalk (Noldus)15 apparatus are generally used to assess motor coordination in rats.

Protocols are presented for two established motor coordination tasks, the accelerating rotarod and horizontal bar, also two tests developed in Oxford recently, the static rods and parallel bars. These tests can detect motor impairments potentially of interest in their own right, as well as being possible variables in tests of other areas of behavior.

测量小鼠运动协调

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such ...

科研

神经科学

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life. The cold plantar assay allows the objective and inexpensive assessment of cold sensitivity in mice, and can quantify both analgesia and hypersensitivity. Mice are acclimated on a glass plate, and a compressed dry ice pellet is held against the glass surface underneath the hindpaw. The latency to withdrawal from the cooling glass is used as a measure of cold sensitivity.
Cold sensation is also important for survival in regions with seasonal temperature shifts, and in order to maintain sensitivity animals must be able to adjust their thermal response thresholds to match the ambient temperature. The Cold Plantar Assay (CPA) also allows the study of adaptation to changes in ambient temperature by testing the cold sensitivity of mice at temperatures ranging from 30 &#176;C to 5 &#176;C. Mice are acclimated as described above, but the glass plate is cooled to the desired starting temperature using aluminum boxes (or aluminum foil packets) filled with hot water, wet ice, or dry ice. The temperature of the plate is measured at the center using a filament T-type thermocouple probe. Once the plate has reached the desired starting temperature, the animals are tested as described above.
This assay allows testing of mice at temperatures ranging from innocuous to noxious. The CPA yields unambiguous and consistent behavioral responses in uninjured mice and can be used to quantify both hypersensitivity and analgesia. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

The Cold Plantar Assay (CPA) measures cold responsiveness between 30 &#176;C and 5 &#176;C, and can also measure cold adaptation. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

测定冷灵敏度和适应小鼠一种简单廉价的方法

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

Assessment of Dopaminergic Homeostasis in Mice by Use of High-performance Liquid Chromatography Analysis and Synaptosomal Dopamine Uptake

Dopamine (DA) is a modulatory neurotransmitter controlling motor activity, reward processes and cognitive function. Impairment of dopaminergic (DAergic) neurotransmission is strongly associated with several central nervous system-associated diseases such as Parkinson&#39;s disease, attention-deficit-hyperactivity disorder and drug addiction1,2,3,4. Delineating disease mechanisms involving DA imbalance is critically dependent on animal models to mimic aspects of the diseases, and thus protocols that assess specific parts of the DA homeostasis are important to provide novel insights and possible therapeutic targets for these diseases.
Here, we present two useful experimental protocols that when combined provide a functional read-out of the DAergic system in mice. Biochemical and functional parameters on DA homeostasis are obtained through assessment of DA levels and dopamine transporter (DAT) functionality5. When investigating the DA system, the ability to reliably measure endogenous levels of DA from adult brain is essential. Therefore, we present how to perform high-performance liquid chromatography (HPLC) on brain tissue from mice to determine levels of DA. We perform the experiment on tissue from dorsal striatum (dStr) and nucleus accumbens (NAc), but the method is also suitable for other DA-innervated brain areas.
DAT is essential for reuptake of DA into the presynaptic terminal, thereby controlling the temporal and spatial activity of released DA. Knowing the levels and functionality of DAT in the striatum is of major importance when assessing DA homeostasis. Here, we provide a protocol that allows to simultaneously deduce information on surface levels and function using a synaptosomal6 DA uptake assay.
Current methods combined with standard immunoblotting protocols provide the researcher with relevant tools to characterize the DAergic system.

Synaptosomal dopamine uptake and high-performance liquid chromatography analysis represent experimental tools to investigate dopamine homeostasis in mice by assessing the function of the dopamine transporter and levels of dopamine in striatal tissue, respectively. Here we present protocols to measure dopamine tissue content and assess the functionality of the dopamine transporter.

高效液相色谱分析和触多巴胺摄取对小鼠多巴胺能稳态的评价

Dopamine (DA) is a modulatory neurotransmitter controlling motor activity, reward processes and cognitive function. Impairment of dopaminergic (DAergic) ...

An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and parenting. Importantly, mice can be trained to associate novel odors with specific behavioral responses to provide insight into olfactory circuit function. This protocol details the procedure for training mice on a Go/No-Go operant learning task. In this approach, mice are trained on hundreds of automated trials daily for 2&#8211;4 weeks and can then be tested on novel Go/No-Go odor pairs to assess olfactory discrimination, or be used for studies on how odor learning alters the structure or function of the olfactory circuit. Additionally, the mouse olfactory bulb (OB) features ongoing integration of adult-born neurons. Interestingly, olfactory learning increases both the survival and synaptic connections of these adult-born neurons. Therefore, this protocol can be combined with other biochemical, electrophysiological, and imaging techniques to study learning and activity-dependent factors that mediate neuronal survival and plasticity.

Here, we train mice on an associative learning task to test odor discrimination. This protocol also allows for studies on learning-induced structural changes in the brain.

小鼠嗅觉学习和鉴别的客观和可重复性试验

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and ...

Transcranial Direct Current Stimulation (tDCS) in Mice

Transcranial direct current stimulation (tDCS) is a non-invasive neuromodulation technique proposed as an alternative or complementary treatment for several neuropsychiatric diseases. The biological effects of tDCS are not fully understood, which is in part explained due to the difficulty in obtaining human brain tissue. This protocol describes a tDCS mouse model that uses a chronically implanted electrode allowing the study of the long-lasting biological effects of tDCS. In this experimental model, tDCS changes the cortical gene expression and offers a prominent contribution to the understanding of the rationale for its therapeutic use.

Transcranial Direct Current Stimulation tDCS in Mice

Transcranial direct current stimulation (tDCS) is a therapeutic technique proposed to treat psychiatric diseases. An animal model is essential for understanding the specific biological alterations evoked by tDCS. This protocol describes a tDCS mouse model that uses a chronically implanted electrode.

经颅直流电刺激 (tDCS) 在小鼠中的作用

Transcranial direct current stimulation (tDCS) is a non-invasive neuromodulation technique proposed as an alternative or complementary treatment for ...

Quantitative Measurement of Intrathecally Synthesized Proteins in Mice

Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the CSF protein composition delivers crucial information in basic neuroscience research as well as neurological diseases. One caveat is that proteins measured in CSF may derive from both intrathecal synthesis and transudation from serum, and protein analysis of CSF can only determine the sum of these two components. To discriminate between protein transudation from the blood and intrathecally produced proteins in animal models as well as in humans, CSF protein profiling measurements using conventional protein analysis tools must include the calculation of the albumin CSF/serum quotient (Qalbumin), a marker of the integrity of the blood-brain interface (BBI), and the protein index (Qprotein/Qalbumin), an estimate of intrathecal protein synthesis. This protocol illustrates the entire procedure, from CSF and blood collection to quotients and indices calculations, for the quantitative measurement of intrathecal protein synthesis and BBI impairment in mouse models of neurological disorders.

Elevated spinal fluid protein levels can either be the result of diffusion of plasma protein across an altered blood-brain barrier or intrathecal synthesis. An optimized testing protocol is presented in this article that helps to discriminate both cases and provides quantitative measurements of intrathecally synthesized proteins.

小鼠体内合成蛋白的定量测量

Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the ...

Electrocorticographic Recording of Cerebral Cortex Areas Manipulated Using an Adeno-Associated Virus Targeting Cofilin in Mice

The use of electrocorticographic (ECoG) recordings in rodents is relevant to sleep research and to the study of a wide range of neurological conditions. Adeno-associated viruses (AAVs) are increasingly used to improve understanding of brain circuits and their functions. The AAV-mediated manipulation of specific cell populations and/or of precise molecular components has been tremendously useful to identify new sleep regulatory circuits/molecules and key proteins contributing to the adverse effects of sleep loss. For instance, inhibiting activity of the filamentous actin-severing protein&#160;cofilin&#160;using AAV prevents sleep deprivation-induced memory impairment. Here, a protocol is&#160;described that combines the manipulation of cofilin function in a cerebral cortex area with the recording of ECoG activity to examine whether cortical cofilin modulates the wakefulness and sleep ECoG signals. AAV injection is performed during the same surgical procedure as the implantation of ECoG and electromyographic (EMG) electrodes in adult male and female mice. Mice are anesthetized, and their heads are shaved. After skin cleaning and incision, stereotaxic coordinates of the motor cortex are determined, and the skull is pierced at this location. A cannula prefilled with an AAV expressing cofilinS3D, an inactive form of cofilin, is slowly positioned in the cortical tissue. After AAV infusion, gold-covered screws (ECoG electrodes) are screwed through the skull and cemented to the skull with gold wires inserted in the neck muscles (EMG electrodes). The animals are allowed three weeks to recover and to ensure sufficient expression of cofilinS3D. The infected area and cell type are verified using immunohistochemistry, and the ECoG is analyzed using visual identification of vigilance states and spectral analysis. In summary, this combined methodological approach allows the investigation of the precise contribution of molecular components regulating neuronal morphology and connectivity to the regulation of synchronized cerebral cortex activity during wakefulness and sleep.

This article describes a protocol for the manipulation of molecular targets in the cerebral cortex using adeno-associated viruses and for monitoring the effects of this manipulation during wakefulness and sleep using electrocorticographic recordings.

使用腺相关病毒在小鼠中瞄准科菲林操纵的大脑皮层区域的电皮质记录

The use of electrocorticographic (ECoG) recordings in rodents is relevant to sleep research and to the study of a wide range of neurological conditions. ...

小鼠运动皮层中的细胞外电生理记录

Multichannel Extracellular Recording in Freely Moving Mice

The protocol aims to uncover the properties of neuronal firing and network local field potentials (LFPs) in behaving mice carrying out specific tasks by correlating the electrophysiological signals with spontaneous and/or specific behavior. This technique represents a valuable tool in studying the neuronal network activity underlying these behaviors. The article provides a detailed and complete procedure for electrode implantation and consequent extracellular recording in free-moving conscious mice. The study includes a detailed method for implanting the microelectrode arrays, capturing the LFP and neuronal spiking signals in the motor cortex (MC) using a multichannel system, and the subsequent offline data analysis. The advantage of multichannel recording in conscious animals is that a greater number of spiking neurons and neuronal subtypes can be obtained and compared, which allows the evaluation of the relationship between a specific behavior and the associated electrophysiological signals. Notably, the multichannel extracellular recording technique and the data analysis procedure described in the present study can be applied to other brain areas when conducting experiments in behaving mice.

作者聚焦:用于研究自由移动小鼠神经元活动的多通道细胞外记录的进展 

The protocol describes the methodology of extracellular recording in the motor cortex (MC) to reveal extracellular electrophysiological properties in freely moving conscious mice, as well as the data analysis of local field potentials (LFPs) and spikes, which is useful for evaluating the network neural activity underlying behaviors of interest.

自由移动小鼠的多通道细胞外记录

The protocol aims to uncover the properties of neuronal firing and network local field potentials (LFPs) in behaving mice carrying out specific tasks by ...