Name: morris water maze test: optimization for mouse strain and testing environment
Uploaded: 2015-06-22T13:00:00.000+00:00
Duration: 10 min 24 s
Description: Watch this Scientific Journal Video about Morris Water Maze Test: Optimization for Mouse Strain and Testing Environment at JoVE.com

这项工作是支持普通医学科学研究所（里德/恩格勒-Chiurazzi - U54GM104942），国立神经疾病和中风（阿什 - R01NS33249，R01NS63249和R01NS79374），铜业（恩格勒-Chiurazzi - P20GM109098）时，阿尔茨海默氏症协会（里德 - NIRG-12-242187），西弗吉尼亚大学一个研究学部参议院拨款（里德），一个WVU PSCOR补助金（里德），以及内部资金从医学教务处的西弗吉尼亚大学学院（恩格勒-Chiurazzi）。内容完全是作者的责任，并不一定代表美国国立卫生研究院或阿尔茨海默氏症协会的官方意见。

所有的实验程序按照机构动物护理和使用委员会（IACUC）的标准进行，并经西弗吉尼亚大学的IACUC。 1.前处理<ol><li>成立了池<ol><li>丙预处理池，以便它被提升到一个舒适的高度。 </li><li>放置2升的水（21℃）在预处理池到大约为1厘米的水平。 注意:不要添加色素。 </li></ol></li><li>步骤<ol><li>从动物饲养到检测室带来的老鼠。 </li><li>验证主体识别（主题号，尾纹身，耳夹， 等 ）。 </li><li>标记的小鼠的尾巴用永久性标记来已经经过预先处理从那些仍然待测试的测试笼中区分小鼠。标志着在一个笼子里所有的小鼠用相同的颜色。 </li><li>将鼠标为approxima转移烧杯tely 5秒。然后，轻轻倾鼠标移出烧杯到预处理池。 </li><li>使用计时器，让鼠标留在池中，持续20秒。 </li><li>经过20秒，轻轻地放在瓢（不钓鱼吧）在鼠标前端。简单地说让鼠标探索瓢，如果需要，鼓励鼠标轻轻滑动舀下鼠标，同时慢慢抬起来获得的独家新闻。小心不要吓唬老鼠的独家新闻。 </li><li>当鼠标处于瓢，运回控股笼。 </li><li>在笼子10天重复这些过程对每个小鼠每天一次。定期（ 例如每个动物后），拿起粪便勃利和床上用品使用的净池。 </li><li>用清水冲洗定期（ 例如在完成小鼠的笼以下）转印烧杯中。 </li><li>一旦这些步骤完成后，用清水冲洗几次转移烧杯中，dump出来的水池，并冲洗池大约两倍水。 </li></ol></li></ol> 2.可视化平台培训<ol><li>设置鼠标性能跟踪软件如浏览器（ 见表材料 ）。 <ol><li>校准视频相匹配的迷宫的尺寸下的"配置" - "过滤器和对象"选项卡。 </li><li>设置过滤器，使该软件可以区分背景鼠标。 <ol><li>调节"灵敏度"的"后台滤波器"区域下方，使得跟踪软件识别动物。 </li><li>点击"接受"选项，一旦跟踪软件识别的动物。选择"编辑"选项迷宫区以外的地区停电。 </li><li>选择适当的"动物过滤器"和"分。动物的大小"。 </li></ol></li></ol></li><li>创建运行表&lt;OL&gt; <li>创建运行表列出每个鼠标，每个可能的位置，并包含有关的异常行为情况说明的空间。 </li><li>预先确定为可见平台测试都是相同的每组小鼠的位置。因此，对于第一次试验，预确定该平台将被放置在相对鼠标位置被放置到迷宫，右侧靠近壁的一侧。对于第二个试验中，预先确定，该平台将被放置在不同的位置沿后壁每只小鼠并认为这将继续对每个6的试验。 </li></ol></li><li>成立房<ol><li>在房间里坑的窗帘，使得任何空间线索被遮掩。 </li></ol></li><li>成立了浴池。 <ol><li>将浴缸所以它通过确保迷宫的跟踪软件配置匹配上的"配置"配置设置相匹配 - "区域定义"选项卡。注:^ hELPS确保桶是在适当的位置之前，以水填充它作为桶不容易重新安置的一次填补。 </li><li>填用自来水桶，并用温度计，确保水温度大约为21℃。确保水保持在此温度下的一个度在整个测试，较冷的水可以用老年小鼠8时影响性能更是如此。 注意:不要添加色素。 </li><li>使用的预定位置，将平台，它有一个安装标志，达到13厘米的高度和4.5厘米×4.5厘米用黑体的"S"形的字符浮雕在其上，在桶的第一个位置，以便它大约为1英寸的水的表面上方。 </li></ol></li><li>步骤<ol><li>就拿第一个鼠标从笼子里，验证主体识别，标记使用正确颜色的记号笔在烧杯尾巴，和地点。 </li><li>轻轻将谅解备忘录e取入浴盆，启动计时器，并确保跟踪软件跟踪动物。让鼠标达2分钟，以定位在平台。一旦鼠标定位的平台，让鼠标20秒的平台上。 </li><li>如果动物找到平台，并进行到跳或脱落，舀鼠标和将鼠标在平台上。如果鼠标继续跳或脱落的平台，按住鼠标的平台上，因此，老鼠得知其余在平台上线从浴缸中逃生。 注意:鼠标应保持20秒的一个组合时的平台上;因此，如果鼠标5秒后跳下，左平台上的时间量为15秒，没有重置为20秒。 </li><li>如果动物没有发现在2分钟内的平台，舀鼠标，并将其放置在平台上，持续20秒。 </li><li>在平台上以下20秒，从平台移开鼠标为已完成预处理训练和将鼠标在第Ë加热笼内衬用纸巾并加热到〜31°C通过加热垫和热灯30秒。与直肠温度计进行体温的周期性评估，以确保小鼠或小鼠的某些菌株在某年龄都没有差异易受低温诱导暴露于水。 </li><li> 2分钟后，以保持笼与加热灯，鼠标转移至保持笼子只用加热垫，但没有热灯，试验之间恢复。最后，把从单一的笼子全部小鼠在同一个笼子里举行测试期间。 </li><li>净清洁碎片的迷宫后，每鼠已经完成了试验，扰乱嗅觉线索。 </li></ol></li><li>每个鼠标组中的重复这些步骤。后每只小鼠已完成第一次试验中，将平台向第二预定位置。 注意:该平台将单个试验期间保持在相同的位置为每个小鼠和将移动到另一预定位置为随后的试验。这样做是为了确保老鼠1）学习有一个平台，2）直接游到可视化平台，从而表现出完整的视觉能力，逃脱和3）没有运动障碍。 </li><li>用于测试的前3天执行这些步骤6的审判。因此，在测试天1，2，和3中执行可见训练，用大约10（ 图2）每次试验之间分钟。 </li><li>在可视化平台的培训，使用动物跟踪软件，可以衡量延迟到达平台，追踪动物的路径长度，和游泳速度。从到Excel电子表格中的"水迷宫"下的"实验列表"，并直接出口标签获得这些测量。 注意:使用一个手计时器也可以是有益的，以确保跟踪软件的准确性。 </li></ol> 3. Morris水迷宫隐藏平台培训<ol><li> S等了房间<ol><li>挂上窗帘在房间模糊空间线索在实验室。 </li><li>将线索战略性的窗帘。确保线索都很大，包括对比的颜色（例如:黑色和白色）以获得更好的可视性。杭线索在距离和高度，他们都是可见的从桶内部小鼠。注意:线索是静态的，在测试过程中不移动。 </li></ol></li><li>设置鼠标性能跟踪软件<ol><li>校准视频相匹配的迷宫的尺寸下的"配置" - "过滤器和对象"选项卡。 </li><li>设置的过滤器，以使软件能够区分从背景的小鼠。 <ol><li>使用相同的过程在可视化平台测试说明（步骤2.1.2.1-2.1.2.3）来设置过滤器Morris水迷宫隐藏平台的训练。 </li></ol></li><li>创建迷宫里面四个相等象限<ol><li>点击"区域定义"选项卡下的"椭圆"选项第一，并创建一个圆圈在屏幕上的迷宫相匹配。注:在此选项卡，将显示实际的迷宫成立，因此它允许选择相匹配的计算机设置与实际迷宫设置。 </li><li>点击"矩形"选项，创建四个相等的正方形。在画面上放置这些方块到新创建的圈子内创建了四个象限相等。 </li></ol></li><li>创建在迷宫的平台。 <ol><li>选择"椭圆"选项，并创建一个圆，将用于该平台的大小。注意:放置在平台中的迷宫事先可以是有益的，以创建一个椭圆的平台的确切大小。 </li><li>将新创建的平台位置在目标象限的中心。 （建议命名这个对象的"目标"，从休息区分开正确的平台）。关键的一步:在框中只是BEL嗷嗷"网格"，选择在"触发"目标平台"停止"选项。这将导致程序停止，一旦鼠标到达平台。 </li><li>创建三个其它相同的平台区，并将其放置在其他象限的确切位置。在"触发器"选项，不要选择"停止"。 </li></ol></li></ol></li><li>创建运行表<ol><li>创建run片列出每个小鼠，每次试验，一个空间写 ​​下到达平台的时间，和一个空间地注意到异常行为（ 表1）。 </li><li>预确定每只小鼠以伪随机方式释放点。使用伪随机选择，使得到该平台的距离等于每一天，每个的4处被同等使用，角度从开始到平台（ 即 ，从左侧或右侧）的内和天之间的平衡。 注意:该平台的位置上跑片，以确保平台的位置不从原始位置移动。确保平台处于所述跟踪软件创建的四个区域中的一个的中心。 注意:如果纵向测试是在不同年龄正在进行，移动台到一个新的位置，在每个年龄。 </li></ol></li><li>成立了迷宫<ol><li>无论是调节水迷宫或相机等，有迷宫，并在"配置"配置设置之间的匹配 - "区域定义"选项卡。 </li><li>标签迷宫（N，S，E，W）的四个象限看不见。检查这些匹配中的跟踪软件，以确保小鼠的配置被从正确的起点释放。 临界步骤:将迷宫外进出鉴于考虑到在迷宫中的线索构成非空间的，非海马依赖性任务游泳动物这些标签。 </li><li>填充用自来水水:迷宫R（约21℃），以使平台是水的表面下大约5毫米。 </li><li>放置在平台中的预定位置。 注:该平台的位置将保持不变在所有天，试验正在测试所有小鼠。 </li><li>使用无毒的白色油漆彩画，使水不透明。这样做是为了确保平台的顶部是从动物的眼睛水平，而游泳无形。 </li></ol></li><li>步骤<ol><li>就拿鼠标从笼子里，用相应颜色的记号笔标注的尾巴，并放置在烧杯中。 </li><li>轻轻倾鼠标进入迷宫，使其进入面壁。注意:每个鼠标将被放置在相同的起始位置为一个唯一的试验。 </li><li>在鼠标的最初版本，开始计时，并站在一个领域的测试是不容易看到的老鼠。确保动物跟踪软件跟踪正常的动物。 </l我&gt; <li>一旦动物到达平台，使其保持在平台上15秒。 注意:这允许动物定向到房间内的空间位置。 </li><li>经过15秒的平台上，从迷宫中取出的动物，并返回鼠标加热笼。 </li><li>如果鼠标发现平台，并进行到跳跃或前15秒脱落，舀小鼠并将其放回为15秒的剩余部分的平台上，以便鼠标学会与逃生平台相关联。 </li><li>如果鼠标在60秒内找到平台，轻轻舀老鼠，并将其放置在平台上。使其保持在平台上15秒，然后用勺子取出动物，并把它返回到加热的笼中。 </li><li>清理杂物的迷宫净扰乱嗅觉线索。 </li><li>对于该组中的其余小鼠重复这些过程。因此，对于测试的4天到9，进行隐蔽站台四个试验每天荷兰国际集团与每次试验之间大约20分钟。 </li><li>更改起始位置pseduorandomly每个审判。因此，试验1中，释放所有的小鼠从相同的起始位置，然后从用于随后的试验不同的起始位置重复该过程。每一天，不同的发行点，以确保动物没有制定一个非空间，非海马依赖肌肉运动的策略。例如，如果小鼠选自N，S，E和W点释放在第一天，不释放它们按照相同的顺序翌日。 </li><li>期间隐藏平台训练，使用动物跟踪软件来测量延迟到达平台，该动物的路径长度，以及时间的百分比或距离的动物花费在每个迷宫象限。获得"水迷宫"选项卡这些测量下的"实验列表"，可以直接导出到Excel电子表格。 注意:使用一个手定时器的保证了跟踪的准确性软件。 </li></ol></li></ol> 4.探测试验<ol><li>设置鼠标性能跟踪软件<ol><li>使用相同的步骤，在Morris水迷宫隐藏平台过程（步骤3.2.1-3.2.4.3）有一个例外。关键的一步:对于探头试验，确保"一站式"选项"触发"期间探头审判被关断。这将防止从停止当鼠标穿过平台区的程序。 </li></ol></li><li>创建运行表<ol><li>创建运行表列出每个鼠标，每次试验和空间要注意异常行为。在探针试验预先确定的释放点，使得它们交替两侧的平台相对之间。因此，预确定使得小鼠不从两侧是邻近于平台释放的释放点。 </li></ol></li><li>步骤<ol><li>进行类似的隐藏平台培训的探头试验，与excep针对这样没有平台是在迷宫中。 </li><li>从预定的释放点释放鼠标。在探测试验，去除平台和鼠标有60秒的迷宫游泳。 </li><li>在测试过程中，进行4探头试验。改变探针试验间的小鼠的释放点。因此，从以前的探测试验的改变目前的探测试验的释放点。 </li><li>在探针的试验中，使用动物跟踪软件来跟踪动物的路径长度，时间的百分比的动物花费在迷宫的每个象限，和动物游在先前platformed区域的次数。从"数据分析"选项卡，在"实验列表"中的"水迷宫"选项卡，然后到Excel电子表格直接出口获得这些测量。 注意:使用一个手计时器也可以是有益的，以确保跟踪软件的准确性。 </li><li>之前进行探测试验隐藏平台训练天6，7，8和10（ 图2）。这些探测试验后，进行隐蔽平台4项试验的每一天的训练。因此，执行探测第一，紧接着使用相同程序的所有小鼠隐藏的平台培训在第3节中描述相反，请在10天的探测试验后不进行额外的隐藏平台的训练。 </li></ol></li></ol> 5.分析<ol><li>可见平台培训<ol><li>进行重复测量的方差分析分别为（一）的路径长度，（二）延迟找到平台，以及（c）游泳速度，用转基因（或其他变量，如治疗）作为变量和天或训练块从属之间作为在被试内变量。注意:对于更复杂的设计，与统计学家咨询可能是有用的。关于组的信息，请参阅8,13尺寸足以适当的开机这项任务与阿尔茨海默常用机型7病。 </li><li>无能表现<ol><li>识别性能-无能小鼠，包括小鼠，演示视觉或电机无能，或动物不获得测试的程序组件，和从隐藏平台和探头性能的随后的统计分析中删除这些小鼠。 <ol><li>计算平均潜伏期和游泳速度为每个鼠标可视化平台培训的最后一天。删除小鼠装置2组以上标准偏差的意思，因为这可能表明电动机或视觉障碍的。 </li><li>识别和清除小鼠没有定向或跟随逃跑瓢，或表现出小鼠的异常行为，如软木螺杆游泳或浮动。 </li></ol></li></ol></li></ol></li><li> Morris水迷宫隐藏平台培训<ol><li>进行上（一）光程重复测量的方差分析，（二）延迟到发现隐藏平台，（三）游泳速度，和（d）的时间百分数或百分比距离象限与转基因作为学科之间的变量和试验的受试者内变量。 </li></ol></li><li>探测试验<ol><li>游泳速度<ol><li>在探测器试验的转基因进行的游泳速度重复测量的方差分析的学科之间的变量和试验的受试者内变量。 </li></ol></li><li>平台交叉索引<ol><li> - 等效位置中的其他3个象限平均交叉次数的PCI =数鼠标越过目标位置:使用探针试验的配置对于每个小鼠，平台杂交指数（PCI）用下列公式计算。 注:这是进行以确定是否鼠标使用空间搜索策略，通过多口岸过培训的平台位置，或thigmotaxic游泳的非空间战略所示，通过在四个地点约等于口岸所示。 </li><li>重复行为方差分析的措施对PCI与转基因作为学科之间的变量和试验的受试者内变量。 </li></ol></li><li> ％的时间或象限％的距离<ol><li>计算％的时间或距离每只小鼠花费在四个象限的迷宫。反复进行方差分析的措施上的时间百分比，或距离，用转基因的个体间的变量和试验的受试者内变量。使用百分比的时间来确定一个空间搜索策略是否正在使用;大约25％的时间在目标象限的研究结果表明小鼠正在执行在机会水平和未使用空间搜索策略。 </li></ol></li><li> O / N遗忘<ol><li>在探测试验平台比较渡区的潜伏期延迟定位平台前一天的最后审判。注意:如果在探头试验平台横穿的延迟是显著长于延迟找到platfor隐藏平台的前一天最后的审判过程中m，则O / N遗忘的平台位置发生。 </li></ol></li></ol></li><li> 事后分析<ol><li>按照与转基因的，在每次试验事后比较显著RMANOVAs。 </li></ol></li></ol>重新优化的一种新的测试环境6例<ol><li>预处理。进行预处理，在容纳室。放置在笼子上没有一个运输车在整个会话盖子。 注意:目的用于预处理过程的变形例是更逐步引入的处理程序的操作和暴露于运输装置（烧杯和铲），并增加时间小鼠是在家里笼而不microisolator盖子和以外的笼子中开放的领域和更亮的光线条件。 <ol><li> 1和2天<ol><li>让老鼠来适应环境的手在笼子之中，其次是温婉动人，随后被提升纹身检查和尾部的标记。在出发前处理2天，将补充材料筑巢在笼子里，为筑巢和住所提供额外的材料，而盖子已被删除9。 </li></ol></li><li>天3和4 <ol><li>放置运输烧杯，舀，和在预处理箱的可见平台的标志，并允许小鼠在每次试验2分钟探索2试验。 </li><li>在这个协议的过程中预先处理，不要使用水。相反，确保盒包含足够干净的被褥覆盖一层，香味用少量雌雄弄脏垃圾如果两种性别被测试。每天更换床上用品。 </li></ol></li><li>第5天<ol><li>地方小鼠在预处理盒含有轻度污染垫料和可见平台标志为3的试验，对于每20秒。使用烧杯和舀运送小鼠和从框。 </li></o升&gt; </li></ol></li><li>可见平台培训<ol><li>设定水温为27℃（通常用于明尼苏达测试环境）。 </li><li>返回小鼠家笼时获取所述提示的平台。 </li><li>让老鼠最长为60秒收购可视化平台。 </li><li>开展培训课程包括每天3试验为5天。 </li></ol></li><li>隐藏平台培训<ol><li>修改视觉线索。除了一大幕掩盖处理，无论是消除或使周围的水池窄等窗帘，以便有更开放的空间。放置其他物品在房间里（ 如 ，暴露的货架，黑色面料海报白色符号，黑色和白色的沙滩球，大黑烟囱，笔记本电脑和一个黑色的毛绒动物），导致均衡的，但更多样化的视觉CUE-设置比以前使用的。 </li><li>开始隐藏平台训练72小时以下的可视化平台traini完成NG，随着试验的大约30分钟的间隔。 </li><li>开展培训课程包括2项试验8天。 </li></ol></li><li>探测试验<ol><li>执行探测试验72小时以下隐藏训练试验8，12，和16.Include试验8-9之间为期3天的间隔，以及培训12-13。 </li><li> 30秒配置探测持续时间。 </li></ol></li></ol>

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J., Arseneau, L. M., Beverly, J. L., Juraska, J. M. <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+estrous+cycle+on+water+maze+acquisition+depends+on+the+temperature+of+the+water.">Effect of estrous cycle on water maze acquisition depends on the temperature of the water.</a> Behavioral Neuroscience. 118 (4), 863-868 (2004).</li></ol>

作者什么都没有透露。

在MWM任务被广泛用于评估空间学习和记忆。然而，这个任务的鲁棒性可受许多因素的影响，并需要优化为背景应变和测试环境。如在图4中，相同的训练协议和应用在两个不同的检测室（相当于大小和布局）使用视觉提示产生显著不同的探针性能。由于检测室的许多特征可能有助于空间线索4，据推测，这两个房间中的一些这使测试更困难未知的方式显著不同。在修改的额外迷?...

转基因小鼠模型已经有助于评估阿尔茨海默氏病（AD）的病理生理学，以及治疗性干预的潜在。认知任务，如在Morris水迷宫（MWM），通常使用与这些模型，以确定的记忆障碍的分子相关因素，并评估临床前药物的疗效。这是至关重要的，但是，认知任务的动态范围足够宽，以检测细微的治疗效果。与AD小鼠模型，认知缺陷通常是年龄依赖性，小鼠显示性能逐步下降（ 例如 ，1）。利用一个敏感的认知任务的可允许检测细微的差别较早在动物的寿命，从而减少与衰老动物相关的成本。例如，减少的海马依赖性巴恩斯训练试验的数目从15到5迷宫增加了工作的难度，造成对d在3xTg模型赤字在较早的年龄比以前etection报道2。早期发现的赤字不仅提供了大量的时间和成本节约，这也增加了可能性，潜在的认知缺陷的分子变化可以被识别。 影响的认知任务的灵敏度的一个因素是小鼠模型的遗传背景应变。例如，BALB / c小鼠相比其他菌株，如C57BL / 6 3表现出优异的性能在学习和记忆任务。 F1的FVB /的N×129S6背景用于两个AD的最广泛的机型，和的Tg2576 RTG（TauP301L）4510款。该菌株表现出MWM相对于其他菌株，包括B6 / SJL小鼠4出众的学习能力。正因为如此出众的学习能力，经过大量的训练使用单一探头可以掩盖从过度训练造成的组间差异。此外，sensitivit探测试验y可以为年龄依赖性。之前我们已经表明，较早的探测试验，经过有限的隐藏平台的训练，都是在年轻的Tg2576的差异相比，比年轻的转基因阴性对照同窝更敏感更广泛的培训后，5探头插入试验。相比之下，经过广泛的培训探测试验是中老年（20-25个月）相比老同窝Tg2576小鼠更敏感比更早的探测试验5。由散布探针试验在整个训练，可能是一个敏感的试验将确定增加，尤其是当纵向测试被执行并且一个特定的探针试验的灵敏度是年龄依赖性的。 图1示出的F1 FVB / N x的性能优越对于这株优化协议下129S6小鼠相比B6 / SJL背景下的协议一起训练更广泛的培训小鼠。 该MWM是一般认为，以提供可靠的措施，是在这两个时间和实验室6再现的。例如，主协议最初由美国明尼苏达我们的实验室1,7已成功地与西弗吉尼亚大学8细微的修改来实现。同样，减值相当于水平RTG（TauP301L）的变化相对4510小鼠，如果安置无病原体或常规条件下，9至对照同窝。然而，该测试环境可以影响MWM任务的灵敏度。因素，例如室内照明，排风口，温度梯度和噪声都有助于环境线索4可最终影响性能。当我们的明尼苏达实验室和动物饲养被移动到一个新的建筑物，最多在野生型性能降低38％，观察，基本上减少了任务和检测转基因相关的缺陷的能力的动态范围。这种变化在PErformance发生尽管设计测试室是等价的尺寸和配置的，并使用相同的施加的视觉线索。 A"重新优化"的原始协议被要求增加新的测试环境的MWM任务的动态范围。 这里专为与F1 FVB / N利用原有协议x 129S6背景5描述。因为一些研究表明应力差MWM性能10和预处理相关的所用性能11减轻这种应力诱发赤字，预处理协议被设计为适应小鼠到池之前MWM测试的引入和除去。以下预处理，小鼠经历可见平台训练，其中一个凸起的平台上都标有一个标志。可见平台训练来识别小鼠与涉及感觉异常性能问题。用在描述PROT排除标准ocol部分，性能不称职的小鼠从隐藏的培训平台和探测试验的后续检查中删除。在隐藏平台的训练和探头试验损伤被解释为认知缺陷，因为感觉性能分解出来的数据。可见平台的培训结束后，老鼠开始隐藏平台的训练，其中该平台浸没在水中，并保持在相对位置外部线索相同。在该平台被除去（探针试验）试验是在散布在整个隐藏平台训练以评估额外训练的影响。因为探针的试验发生在每一天的开始，前附加隐蔽平台训练，探针试验测量动物记住以下一个20小时的延迟平台的位置的能力，考虑参考存储器12的量度。最后，方法，使这个原始协议被重新优化时的变化在测试环境打乱了控制性能描述。

<table><tbody><tr><td>Viewer Tracking software</td><td>Biobserve</td><td></td><td>This particular software is not a requirement - there are other tracking systems available</td></tr><tr><td>Pre-handling pool</td><td></td><td></td><td>Dimensions approximately 1 foot wide x 2 feet long x 1.5 feet deep</td></tr><tr><td>Plastic beaker</td><td></td><td></td><td>1 L</td></tr><tr><td>Scoop</td><td></td><td></td><td></td></tr><tr><td>Small net</td><td></td><td></td><td></td></tr><tr><td>Stopwatch</td><td></td><td></td><td></td></tr><tr><td>Non-toxic white tempera paint</td><td></td><td></td><td>Any color paint can be used; however, most tracking software programs require that the paint contrast with the color of the animal.</td></tr><tr><td>Visible platform</td><td></td><td></td><td>Color should contrast that of maze</td></tr><tr><td>Curtain rod</td><td></td><td></td><td></td></tr><tr><td>Curtains</td><td></td><td></td><td></td></tr><tr><td>Circular tub</td><td></td><td></td><td>Usually white in color; approximately 4 feet in diameter</td></tr><tr><td>Hidden platform</td><td></td><td></td><td>Painted same color as the water</td></tr></tbody></table>

morris water maze test: optimization for mouse strain and testing environment

The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, including neurocognitive disorders such as Alzheimer&#8217;s disease. However, the background strain of the mouse model used can have a substantial effect on the observed behavioral phenotype, with some strains exhibiting superior learning ability relative to others. To ensure differences between transgene negative and transgene positive mice can be detected, identification of a training procedure sensitive to the background strain is essential. Failure to tailor the MWM protocol to the background strain of the mouse model may lead to under- or over- training, thereby masking group differences in probe trials. Here, a MWM protocol tailored for use with the F1 FVB/N x 129S6 background is described. This is a frequently used background strain to study the age-dependent effects of mutant P301L tau (rTg(TauP301L)4510 mice) on the memory deficits associated with Alzheimer&#8217;s disease. Also described is a strategy to re-optimize, as dictated by the particular testing environment utilized.

我们已经使用了Morris水迷宫研究β-淀粉样蛋白（Tg2576小鼠）和突变型P301L的tau空间参考存储器上（RTG（TauP301L）4510只小鼠）（ 例如 ，1,5,7,8）。的影响。图3是代表结果报告在我们的研究考察成人发病P301L头表达对学习和记忆8的效果，利用测试环境A.评估电机和视觉功能，小鼠跨可视化平台培训模块，其中每个训练块组成的比较3试验。光程的可视化平台培训的控?...

Watch this Scientific Journal Video about Morris Water Maze Test: Optimization for Mouse Strain and Testing Environment at JoVE.com

Morris Water Maze Test: Optimization for Mouse Strain and Testing Environment

This manuscript describes a Morris water maze (MWM) protocol tailored for use with a commonly used mouse model of Alzheimer's disease. The MWM is widely used in transgenic mouse models. Implementation of a procedure sensitive to the background strain of the mouse model is essential for detecting group differences.

Morris水迷宫实验:优化小鼠品系和测试环境

 The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, ...

morris-water-maze-test-optimization-for-mouse-strain-testing

Universidad San Sebastian

Research

JoVE Journal

Behavior

21.4K 次观看.  West Virginia University.  This manuscript describes a Morris water maze (MWM) protocol tailored for use with a commonly used mouse model of Alzheimer's disease. The MWM is widely used in transgenic mouse models. Implementation of a procedure sensitive to the background strain of the mouse model is essential for detecting group differences. 

视频: Morris Water Maze Test: Optimization for Mouse Strain and Testing Environment

Shallow Water (Paddling) Variants of Water Maze Tests in Mice

When Richard Morris devised his water maze in 19817, most behavioral work was done in rats. However, the greater understanding of mouse genetics led to the mouse becoming increasingly important. But researchers found that some strains of mutant mice were prone to problems like passively floating or diving when they were tested in the Morris water maze11. This was unsurprising considering their natural habitat; rats swim naturally (classically, the "sewer rat"), whereas mice evolved in the dry areas of central Asia.
To overcome these problems, it was considered whether shallow water would be a sufficient stimulus to provide escape motivation for mice. This would also avoid the problems of drying the small creatures with a towel and then putting them in a heated recovery chamber to avoid hypothermia, which is a much more serious problem than with rats; the large ratio of surface area to volume of a mouse makes it particularly vulnerable to rapid heat loss.
Another consideration was whether a more natural escape strategy could be used, to facilitate learning. Since animals that fall into water and swim away from the safety of the shore are unlikely to pass on their genes, animals have evolved a natural tendency to swim to the edge of a body of water. The Morris water maze, however, requires them to swim to a hidden platform towards the center of the maze - exactly opposite to their evolved behavior. Therefore the paddling maze should incorporate escape to the edge of the apparatus. This feature, coupled with the use of relatively non-aversive shallow water, embodies the "Refinement" aspect of the "3 Rs" of Russell and Burch8.
Various types of maze design were tried; the common feature was that the water was always shallow (2 cm deep) and escape was via a tube piercing the transparent wall of the apparatus. Other tubes ("false exits") were also placed around the walls but these were blocked off. From the inside of the maze all false exits and the single true exit looked the same. Currently a dodecagonal (12-sided) maze is in use in Oxford, with 12 true/false exits set in the corners. In a recent development a transparent paddling Y-maze has been tested successfully.

Shallow Water Paddling Variants of Water Maze Tests in Mice

Mice can swim, but many strains appear to find this activity stressful. To overcome this problem mazes have been devised where escape from shallow water is used to motivate behaviour. These have been demonstrated to support learning at least as good as the traditional and widely used Morris water maze.

浅水（杖刑）的变体在小鼠体内的水迷宫测试

When Richard Morris devised his water maze in 19817, most behavioral work was done in rats. However, the greater understanding of mouse genetics led to ...

科研

行为学

Low-stress Route Learning Using the Lashley III Maze in Mice

Many behavior tests designed to assess learning and memory in rodents, particularly mice, rely on visual cues, food and/or water deprivation, or other aversive stimuli to motivate task acquisition. As animals age, sensory modalities deteriorate. For example, many strains of mice develop hearing deficits or cataracts. Changes in the sensory systems required to guide mice during task acquisition present potential confounds in interpreting learning changes in aging animals. Moreover, the use of aversive stimuli to motivate animals to learn tasks is potentially confounding when comparing mice with differential sensitivities to stress. To minimize these types of confounding effects, we have implemented a modified version of the Lashley III maze. This maze relies on route learning, whereby mice learn to navigate a maze via repeated exposure under low stress conditions, e.g. dark phase, no food/water deprivation, until they navigate a path from the start location to a pseudo-home cage with 0 or 1 error(s) on two consecutive trials. We classify this as a low-stress behavior test because it does not rely on aversive stimuli to encourage exploration of the maze and learning of the task. The apparatus consists of a modular start box, a 4-arm maze body, and a goal box. At the end of the goal box is a pseudo-home cage that contains bedding similar to that found in the animal&#x2019;s home cage and is specific to each animal for the duration of maze testing. It has been demonstrated previously that this pseudo-home cage provides sufficient reward to motivate mice to learn to navigate the maze1. Here, we present the visualization of the Lashley III maze procedure in the context of evaluating age-related differences in learning and memory in mice along with a comparison of learning behavior in two different background strains of mice. We hope that other investigators interested in evaluating the effects of aging or stress vulnerability in mice will consider this maze an attractive alternative to behavioral tests that involve more stressful learning tasks and/or visual cues.

The Lashley III maze is a route-learning task that does not rely on aversive stimuli or visual cues. It is thus a highly attractive option for evaluating learning and memory, especially in aging mice or otherwise where stress is a consideration.

低应力路线学习，在老鼠身上使用拉什利三迷宫

Many behavior tests designed to assess learning and memory in rodents, particularly mice, rely on visual cues, food and/or water deprivation, or other ...

神经科学

Quantifying Cognitive Decrements Caused by Cranial Radiotherapy

With the exception of survival, cognitive impairment stemming from the clinical management of cancer is a major factor dictating therapeutic outcome. For many patients afflicted with CNS and non-CNS malignancies, radiotherapy and chemotherapy offer the best options for disease control. These treatments however come at a cost, and nearly all cancer survivors (~11 million in the US alone as of 2006) incur some risk for developing cognitive dysfunction, with the most severe cases found in patients subjected to cranial radiotherapy (~200,000/yr) for the control of primary and metastatic brain tumors1. Particularly problematic are pediatric cases, whose long-term survival plagued with marked cognitive decrements results in significant socioeconomic burdens2. To date, there are still no satisfactory solutions to this significant clinical problem.
We have addressed this serious health concern using transplanted stem cells to combat radiation-induced cognitive decline in athymic rats subjected to cranial irradiation3. Details of the stereotaxic irradiation and the in vitro culturing and transplantation of human neural stem cells (hNSCs) can be found in our companion paper (Acharya et al., JoVE reference). Following irradiation and transplantation surgery, rats are then assessed for changes in cognition, grafted cell survival and expression of differentiation-specific markers 1 and 4-months after irradiation. To critically evaluate the success or failure of any potential intervention designed to ameliorate radiation-induced cognitive sequelae, a rigorous series of quantitative cognitive tasks must be performed. To accomplish this, we subject our animals to a suite of cognitive testing paradigms including novel place recognition, water maze, elevated plus maze and fear conditioning, in order to quantify hippocampal and non-hippocampal learning and memory. We have demonstrated the utility of these tests for quantifying specific types of cognitive decrements in irradiated animals, and used them to show that animals engrafted with hNSCs exhibit significant improvements in cognitive function3.
The cognitive benefits derived from engrafted human stem cells suggest that similar strategies may one day provide much needed clinical recourse to cancer survivors suffering from impaired cognition. Accordingly, we have provided written and visual documentation of the critical steps used in our cognitive testing paradigms to facilitate the translation of our promising results into the clinic.

Cognitive impairment resulting from the radiotherapeutic management of brain tumors represents a clinically intractable condition that adversely impacts quality of life. The capability to critically evaluate potential interventions for ameliorating radiation-induced cognitive decrements ultimately depends on the capability to undertake rigorous quantitative assessments of cognition.

量化颅放疗引起的认知递减

With the exception of survival, cognitive impairment stemming from the clinical management of cancer is a major factor dictating therapeutic outcome. For ...

医学

Morris Water Maze Test for Learning and Memory Deficits in Alzheimer&#39;s Disease Model Mice

The Morris Water Maze (MWM) was first established by neuroscientist Richard G. Morris in 1981 in order to test hippocampal-dependent learning, including acquisition of spatial memoryand long-term spatial memory 1. The MWM is a relatively simple procedure typically consisting of six day trials, the main advantage being the differentiation between the spatial (hidden-platform) and non-spatial (visible platform) conditions 2-4. In addition, the MWM testing environment reduces odor trail interference 5. This has led the task to be used extensively in the study of the neurobiology and neuropharmacology of spatial learning and memory. The MWM plays an important role in the validation of rodent models for neurocognitive disorders such as Alzheimer&#x2019;s Disease 6, 7. In this protocol we discussed the typical procedure of MWM for testing learning and memory and data analysis commonly used in Alzheimer&#x2019;s disease transgenic model mice.

Morris Water Maze Test for Learning and Memory Deficits in Alzheimer's Disease Model Mice

The Morris Water Maze is a behavioral task to test hippocampal-dependent learning and memory. It has been widely used in the study of neurobiology, neuropharmacology and neurocognitive disorders in rodent models.

在阿尔茨海默氏病模型小鼠学习和记忆障碍的Morris水迷宫测试

The Morris Water Maze (MWM) was first established by neuroscientist Richard G. Morris in 1981 in order to test hippocampal-dependent learning, including ...

T-maze Forced Alternation and Left-right Discrimination Tasks for Assessing Working and Reference Memory in Mice

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in rodents. In our laboratory, we evaluated the two types of memory in more than 30 strains of genetically engineered mice using the automated version of this apparatus. Here, we present the modified T-maze apparatus operated by a computer with a video-tracking system and our protocols in a movie format. The T-maze apparatus consists of runways partitioned off by sliding doors that can automatically open downward, each with a start box, a T-shaped alley, two boxes with automatic pellet dispensers at one side of the box, and two L-shaped alleys. Each L-shaped alley is connected to the start box so that mice can return to the start box, which excludes the effects of experimenter handling on mouse behavior. This apparatus also has an advantage that in vivo microdialysis, in vivo electrophysiology, and optogenetics techniques can be performed during T-maze performance because the doors are designed to go down into the floor. In this movie article, we describe T-maze tasks using the automated apparatus and the T-maze performance of &alpha;-CaMKII+/- mice, which are reported to show working memory deficits in the eight-arm radial maze task. Our data indicated that &alpha;-CaMKII+/- mice showed a working memory deficit, but no impairment of reference memory, and are consistent with previous findings using the eight-arm radial maze task, which supports the validity of our protocol. In addition, our data indicate that mutants tended to exhibit reversal learning deficits, suggesting that &alpha;-CaMKII deficiency causes reduced behavioral flexibility. Thus, the T-maze test using the modified automatic apparatus is useful for assessing working and reference memory and behavioral flexibility in mice.

This article presents the protocol of T-maze tests using a modified automated apparatus for assessing the learning and memory functions in mice.

T-迷宫强迫交替和评估工作，并参考在小鼠记忆的左，右的歧视任务

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in ...

Morris Water Maze Experiment

The Morris water maze is widely used to study spatial memory and learning. Animals are placed in a pool of water that is colored opaque with powdered non-fat milk or non-toxic tempera paint, where they must swim to a hidden escape platform. Because they are in opaque water, the animals cannot see the platform, and cannot rely on scent to find the escape route. Instead, they must rely on external/extra-maze cues. As the animals become more familiar with the task, they are able to find the platform more quickly. Developed by Richard G. Morris in 1984, this paradigm has become one of the "gold standards" of behavioral neuroscience.

The Morris water maze is a well-accepted tool used to document the involvement of the hippocampus in a behavioral task.

Morris水迷宫实验

The Morris water maze is widely used to study spatial memory and learning. Animals are placed in a pool of water that is colored opaque with powdered ...

生物学

A Behavioral Test Battery for the Repeated Assessment of Motor Skills, Mood, and Cognition in Mice

Pharmacological and toxicological studies in neurodegeneration require comprehensive behavioral analysis in mice because motor dysfunctions and dysfunctions in mood and cognition are common and often shared symptoms in neurodegenerative diseases. Shown here is a behavioral test battery for motor, mood, and cognition, which can be repeatedly tested in a longitudinal study. This battery assesses the overall behavioral phenotype in mice by examining each domain of behavior with at least two independent well-accepted tests (i.e., open-field test and rotarod test for motor function, social interaction test, elevated plus maze test, and forced swim test for emotional function, and Morris water maze test and novel object recognition test for cognitive function). Therefore, this sensitive and comprehensive test battery is a powerful tool for the study of behavioral alternation in neurodegeneration.

A comprehensive behavioral test battery of motor skills, mood&#8212;including social interaction, depression, and anxiety&#8212;and cognition is designed for the repeated assessment of neurodegeneration-related behavioral changes in mice.

一种用于小鼠运动技能、情绪和认知重复评估的行为测试电池

Pharmacological and toxicological studies in neurodegeneration require comprehensive behavioral analysis in mice because motor dysfunctions and ...

Analysis of Learning and Memory Ability in an Alzheimer's Disease Mouse Model using the Morris Water Maze

A Morris water maze (MWM) experiment forces experimental animals to swim and learn to find a platform hidden in the water. It is widely used in scientific research to assess the learning and memory of animals. Due to the extensive use of the MWM test, visual experimental protocols are essential for researchers. This manuscript uses the latest studies to introduce the protocol of the MWM test. Alzheimer&#8217; Disease (AD) is characterized by a progressive loss of memory and cognitive function. An alternative and complementary treatment used for AD is Manual Acupuncture (MA). To assess the learning and memory ability of AD model mice, the MWM test was conducted. The visible platform trial, hidden platform trial, probe trial, and reversal trial of MWM were used to evaluate spatial learning and memory ability. In the visible platform trial, the swimming speed and escape latency of mice in different groups was not significantly different. In the hidden platform and reversal trials, the AD group showed a long escape latency. The escape latency decreased significantly after the MA treatment. Low platform crossover number and the proportion of time in the SW quadrant in the probe trial increased after the MA treatment (p &#60; 0.05 or p &#60; 0.01). The results of the MWM tests suggest that MA can effectively improve the spatial learning and memory abilities of AD model mice. Rigorous experimental operations provided assurance of the reliability of the results.

Herein, a protocol to conduct the Morris water maze tests to evaluate the ability of learning and memory of Alzheimer&#8217;s Disease model mice and to assess the effect of manual acupuncture for treating them is described.

使用莫里斯水迷宫对阿尔茨海默氏病小鼠模型学习和记忆能力的分析

A Morris water maze (MWM) experiment forces experimental animals to swim and learn to find a platform hidden in the water. It is widely used in scientific ...

针灸慢性缺氧小鼠的认知改善

Acupuncture Treatment in a Mouse Model of Chronic Hypoxia-Induced Cognitive Dysfunction

The treatment of central nervous disorders has consistently posed significant challenges to the medical field. Acupuncture, a non-pharmacological practice rooted in traditional Chinese medicine, entails the insertion of fine needles into precise points on the body and is commonly employed for the management of diverse conditions. Recently, acupuncture has emerged as a promising therapeutic intervention for a range of neurological diseases, including anxiety and respiratory disorders. However, the potential of acupuncture in treating cognitive dysfunction induced by chronic hypoxia has not yet been explored. This paper presents a comprehensive protocol for establishing a mouse model of chronic hypoxia-induced cognitive impairment, administering mild anesthesia, performing acupuncture treatment, and assessing behavioral changes and memory abilities using open field tests and water mazes. The step-by-step protocol provides detailed instructions on accurately locating and positioning acupoints and needles for cognitive improvement. By employing this protocol, researchers can conduct systematic studies to thoroughly evaluate the therapeutic potential of acupuncture for cognitive dysfunction.

作者聚焦:改进增强小鼠针刺研究的麻醉方案

Here, we describe a protocol for implementing mild anesthesia and acupuncture treatment on a chronic hypoxia mouse model and conducting behavioral tests to assess the cognitive alterations post-treatment.

针灸治疗慢性缺氧诱导认知功能障碍小鼠模型

The treatment of central nervous disorders has consistently posed significant challenges to the medical field. Acupuncture, a non-pharmacological practice ...

Morris水迷宫实验:优化小鼠品系和测试环境

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