这项工作是由加拿大卫生研究院研究院（CIHR），汤森家庭，和杰克布朗和家庭阿尔茨海默氏症研究基金会（到WAS）的支持。 WAS是加拿大研究主席在阿尔茨海默氏病的人。 PL是一个NSERC亚历山大格雷厄姆贝尔加拿大研究生奖学金博士后科研奖和迈克尔史密斯健康研究基金会高级研究生助学金的支持。

1。制备<ol><li>设备的准备工作<ol><li>获取与150厘米直径和深度为50cm（图1）的圆形水池。如果使用的是黑色的老鼠，白池应使用;如果使用的是白老鼠，应使用黑色池。 </li><li>安排的房间，被测试的动物无法看到在测试过程中，实验者。这可以用窗帘或房间隔断。 </li><li>在房间里放置的高对比度空间感，和/或对室内游泳池的位置，这将高出水面。 </li><li>将在游泳池直径10cm平台 - 白色为白池，清晰的黑色池有机玻璃。装满水的池中，直到平台高出水面1厘米。让水平衡至室温（22℃）。根据水的温度，这可能需要一至三天，或者可以加入热水，加快了平衡。 </li></ol></li><li>软件的编制<ol><li>在计算机软件的校准池，使相机可以创建从基于像素的信息物理距离信息。分成4个象限池。指定作为一个变量可以改变每次试验区的平台区。创建5个平台子区域 - 在每个象限，并在池的中心之一。保存校准和使用，其余的测试天。 （例如图2）。 </li><li>设置最大试验时间为60秒。如果鼠标的平台，在此之前找到程序的软件平台被发现时停止试验。 </li><li>指定程序开始自动跟踪，当实验者退出试验区。利用任何“反射最小化”选项，你的软件包提供。 </li><li>跟踪路径的长度，逃避潜伏期和时间花费在每个象限。 </li></ol></li></ol> 2。第1天：可视化平台<ol><li>计算机程序<ol><li>跟踪软件装入池校准。 </li><li>创建5个试验，审间的间隔，适合你的实验。编程平台的位置，并开始每次试验不同的方向。请参阅表1为例协议。 </li></ol></li><li>测试过程<ol><li>将小鼠从他们的住房设施的行为房间。保持在一个地区，在那里他们可以没有看到游泳池或空间线索的老鼠。让他们适应新的环境测试前至少30分钟。 </li><li>放置在平台上，以提高其知名度的一个标志。 </li><li>要开始测试，电梯从家里笼鼠标尾部的基础。支持鼠标，你把它的试验区。解除鼠标的尾巴基地，轻轻放入水中鼠标，面向池边。迅速离开试验区。 </li><li>如果鼠标发现截止前60秒的平台，让鼠标的平台上停留5秒钟，然后返回到其家笼。如果鼠标没有找到平台，平台上放置鼠标和允许留20秒，然后返回到其家笼。 </li><li>重复线索小鼠。随后的审讯开始每个不同的平台的位置和方向开始，你已经到你的软件编程。 </li><li>当测试完成后，返回的老鼠，他们的住房设施。小鼠干之前返回到动物设施的正常体温是保证。 </li><li>在翌日准备，从平台中删除的国旗和添加额外的水淹没的平台，在地表以下1厘米池。 </li></ol></li></ol> 3。 2-5：隐藏的平台<ol><li>计算机程序<ol><li>跟踪软件装入池校准。 </li><li>创建5个试验，审间的间隔，适合你的实验。计划的平台位置保持在相同的位置在所有试验和天，但出发方向不同，每次试验中，每一天。 </li></ol></li><li>测试过程<ol><li>黑鼠，无毒，白色粉末温度油漆添加到池中，调匀。使用足够的涂料等，被淹没的平台是从水面不可见。对于白老鼠，与清澈的海水和明确的有机玻璃平台的黑色游泳池应该使用。 </li><li>按照步骤2.2.3至2.2.6。 </li></ol></li></ol> 4。第6天：探头审判<ol><li>计算机程序<ol><li>跟踪软件装入池校准。 </li><li>创建1审判没有平台区，和一开始的方向。从2-5天使用的平台象限最远的起始方向是首选。径长度为60秒。 </li></ol></li><li>测试过程<ol><li>从池中删除的平台。 </li><li>按照步骤2.2.3至2.2.6。 </li></ol></li></ol> 5。数据分析<ol><li>对于每一天，每一个鼠标，平均5个试验，每个测试题目给一个单一的路径长度和逃避潜伏期。正确计算相结合的错误。对于第6天，只需收集路径的长度，逃避潜伏期和时间花费在每个鼠标的平台象限。 </li><li>如果组第1天之间存在的任何分歧，它可能是一个问题，而不是学习和记忆的视野。只有1天进行分析，如果没有差异。 </li><li>比较天使用适合您的数据集的统计数据2-5的学习曲线。一个陡峭的曲线代表更快的任务收购;浅的曲线表示在收购任务的赤字。从2天到5天的数据分析使用ANOWA。 </li><li>第6天，比较％的时间花费在以前学过的平台象限，使用适合你的数据集的统计数据。一个较高的百分比在平台象限所花费的时间将被解释为一个更高级别的内存保留。 </li></ol> 6。代表性的成果我们用Morris水迷宫测试，以检查AD的发病机制（7）和丙戊酸（VPA）的转基因AD模型小鼠的治疗AD（6）医药潜力缺氧的效果。图3是代表性的结果，我们在我们丙戊酸钠的记忆障碍的影响，在APP23 AD小鼠模型（6）研究报告。在第1天（可视化平台试验），没有VPA治疗组和对照组（图3A）在延迟和路径长度（图3A），表明这两个组也有类似的电机和可视化功能之间的差异。我们从这个假设的老鼠能够看到标记的平台和周围环境的线索，可以游泳，可以接受的。 2-5天（隐藏的平台试验的第1天至4）的例子显示了一个在逃避潜伏期（图3C），组与组之间的路径长度（图3D）的区别，表明丙戊酸钠治疗小鼠的表现明显比对照组更好随着时间的推移。探头线索的最后一天（6日）显示结果，多次前往小鼠进入第三象限，是以前被隐藏的平台，与丙戊酸钠治疗控制（图3E）相比显着。这些数据表明，丙戊酸钠治疗显着提高APP23小鼠的记忆中看到的赤字。 <img src="/files/ftp_upload/2920/2920fig1.jpg" alt="图1"/> 图1。 Morris水迷宫可见平台测试当天的设备安装 。池屏蔽从实验者使用房间隔断。位于空间线索的墙壁上，可能放置在室内池，水面以上，如果需要的话。池充满了清澈的水，表面上方1厘米平台。一个标志，一直放在提升知名度的平台。 <img src="/files/ftp_upload/2920/2920fig2.jpg" alt="图2"/> 图2。屏幕捕获的任何迷宫™视频跟踪，展示池校准系统 。池是从上面看，一个RTV24数字转换器的模拟跟踪摄像机由黑色和白色。几个区域内的软件定义和划分成4个象限的总池。第五，平台区的输入可以各不相同试验的5个可能的地点：西北，东北，西南，东南，或者中心。添加一个校准线（打勾跨越中心线），让软件将其转换成物理距离的像素距离。 <img src="/files/ftp_upload/2920/2920fig3.jpg" alt="图3"/> 图3。 Morris水迷宫代表性的结果。，7个月APP23携带人类瑞典突变APP基因的转基因小鼠每天VPA（N = 30只小鼠）或车辆溶液（N = 30只小鼠）注射一个月后进行了测试。（a ）在的可视化平台测试的第一天，丙戊酸钠治疗和控制APP23小鼠表现出类似的延迟逃脱的可视化平台上。 （二）P &lt;0.05学生的t检验。VPA的治疗和控制APP23小鼠收到类似的游泳距离在可视化平台测试的可视化平台上逃跑。 P&gt; 0.05由学生的t检验。隐藏的平台测试，丙戊酸钠治疗APP23小鼠一个较短的延迟，以逃脱的3 号和第 4天，P到的隐藏平台&lt;由方差0.001。（四）丙戊酸钠治疗APP23小鼠面前逃避到隐藏平台上的第3 次和第 4天，P &lt;0.01由方差一个游泳的长度较短（E）在第6天的探测试验，丙戊酸钠治疗APP23小鼠前往进入第三象限，是以前被隐藏的平台，更多的时间比对照组显着。 * P &lt;0.005学生的t检验。 （改编并从205医学实验 ，2781年至2789年，2008年，美国洛克菲勒大学出版社出版的医学期刊在J.进出口， 杂志转载。10.1084/jem.20081588）（6）。 <table border="1"><tbody><tr><td>&nbsp;</td><td colspan="2"> 第1天 </td><td> 第2天 </td><td> 第3天 </td><td> 第4天 </td><td> 第5天 </td><td> 第6天 </td></tr><tr><td></td><td> 平台的位置 </td><td> 起始方向 </td><td colspan="4"> 平台位置：西南启动位置如下： </td><td> 没有平台。 </td></tr><tr><td> 审判1 </td><td>申银万国</td><td>小号</td><td> W </td><td> ñ </td><td> ñ </td><td> Ë </td><td> ñ </td></tr><tr><td> 审判2 </td><td>净重</td><td> ñ </td><td>小号</td><td> W </td><td> Ë </td><td>小号</td><td></td></tr><tr><td> 试用3 </td><td>东北</td><td>小号</td><td> ñ </td><td> Ë </td><td> W </td><td> W </td><td></td></tr><tr><td> 审判4 </td><td>中心</td><td> Ë </td><td> Ë </td><td> W </td><td>小号</td><td> Ë </td><td></td></tr><tr><td> 试验5 </td><td>东南</td><td> W </td><td>小号</td><td>小号</td><td> ñ </td><td> ñ </td><td></td></tr></tbody></table> 表1。样品水迷宫协议* *请注意平台的位置，第1天开始改变方向，而2-5天的平台的地位保持不变，而出发的方向变化。第6天，有没有平台和审单。第6天开始的方向是最远从以前的平台位置（SW），使小鼠必须旅行之前进入以前学过的平台象限的一段距离。

<ol>
	<li>Morris, R. G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+localization+does+not+require+the+presence+of+local+cues.">Spatial localization does not require the presence of local cues.</a> Learning and Motivation. 12, 239-260 (1981).</li><li>O'Keefe, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+the+hippocampal+place+cells.">A review of the hippocampal place cells.</a> Prog Neurobiol. 13, 419-439 (1979).</li><li>Scoville, W. B., Milner, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Loss+of+recent+memory+after+bilateral+hippocampal+lesions.">Loss of recent memory after bilateral hippocampal lesions.</a> J Neurol Neurosurg Psychiatry. 20, 11-21 (1957).</li><li>Eichenbaum, H., Stewart, C., Morris, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hippocampal+representation+in+place+learning.">Hippocampal representation in place learning.</a> J Neurosci. 10, 3531-3542 (1990).</li><li>Block, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Global+ischemia+and+behavioural+deficits.">Global ischemia and behavioural deficits.</a> Progress in Neurobiology. 58, 279-295 (1999).</li><li>Qing, H., He, G., Ly, P. T., Fox, C. J., Staufenbiel, M., Cai, F., Zhang, Z., Wei, S., Sun, X., Chen, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Valproic+acid+inhibits+Abeta+production,+neuritic+plaque+formation,+and+behavioral+deficits+in+Alzheimer's+disease+mouse+models.">Valproic acid inhibits Abeta production, neuritic plaque formation, and behavioral deficits in Alzheimer's disease mouse models.</a> J Exp Med. 205, 2781-2789 (2008).</li><li>Sun, X., He, G., Qing, H., Zhou, W., Dobie, F., Cai, F., Staufenbiel, M., Huang, L. E., Song, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypoxia+facilitates+Alzheimer's+disease+pathogenesis+by+up-regulating+BACE1+gene+expression.">Hypoxia facilitates Alzheimer's disease pathogenesis by up-regulating BACE1 gene expression.</a> Proc Natl Acad Sci U S A. , 18727-18732 (2006).</li><li>D'Hooge, R., De Deyn, P. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applications+of+the+Morris+water+maze+in+the+study+of+learning+and+memory.">Applications of the Morris water maze in the study of learning and memory.</a> Brain Research Reviews. 36, 60-90 (2001).</li><li>Brandeis, R., Brandys, Y., Yehuda, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+the+Morris+Water+Maze+in+the+study+of+memory+and+learning.">The use of the Morris Water Maze in the study of memory and learning.</a> Int J Neurosci. 48, 29-69 (1989).</li><li>Lipp, H. P., Wolfer, D. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetically+modified+mice+and+cognition.">Genetically modified mice and cognition.</a> Curr Opin Neurobiol. 8, 272-280 (1998).</li><li>Sandi, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+and+mechanisms+of+action+of+glucocorticoid+involvement+in+memory+storage.">The role and mechanisms of action of glucocorticoid involvement in memory storage.</a> Neural Plast. 6, 41-52 (1998).</li></ol>

所有实验均在与不列颠哥伦比亚省的动物护理和使用委员会和CIHR指引大学的规定进行。

年龄，性别，品种，和应变的差异影响的MWM性能（8）。研究表明，老年小鼠在MWM表现不佳，而男性的啮齿类动物的表现比女性更好;此外，浮动比老鼠（9，10）的小鼠更明显。因此，这些元素应等同于在所有的测试。证据还表明，强调动物的MWM（11）执行更差，因此环境因素可能会造成压力，如温度，光照和噪音，，应监测和保持常数了任务。

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>评论（可选） </th></tr><tr><td> AnyMaze视频跟踪系统</td><td> Stoelting公司</td><td></td></tr><tr><td>彩画漆</td><td>里夫斯＆普尔组</td><td>白，粉</td></tr></tbody></table>

morris water maze test for learning and memory deficits in alzheimer&#39;s disease model mice

神经学家理查德G.莫里斯Morris水迷宫（MWM）最初成立于1981年，以测试海马依赖的学习，包括收购空间memoryand长期空间记忆 1 。 MWM的是一个相对简单的的程序通常包含进行为期6天的试验中，之间的空间（隐藏平台的主要优势）的分化和非空间（可视化平台）的条件下 2-4 。此外，MWM的测试环境，减少气味径干扰 5 。这导致了广泛的任务要使用的空间学习和记忆的神经生物学和神经药理学的研究。的MWM中起着重要作用，在啮齿类动物模型的神经认知障碍，如阿尔茨海默氏病6，7验证。在这个协议中，我们讨论的MWM测试学习记忆和阿尔茨海默氏症的转基因小鼠模型中常用的数据分析的典型过程。

Watch this Scientific Journal Video about Morris Water Maze Test for Learning and Memory Deficits in Alzheimer's Disease Model Mice at JoVE.com

Morris Water Maze Test for Learning and Memory Deficits in Alzheimer&#039;s Disease Model Mice

Morris水迷宫测试海马依赖的学习和记忆是一种行为的任务。它已被广泛应用在神经生物学，神经和神经紊乱的啮齿动物模型的研究。

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 ...

morris-water-maze-test-for-learning-memory-deficits-alzheimer-s

Research

JoVE Journal

Neuroscience

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

125.7K 次观看.  University of British Columbia. Morris水迷宫测试海马依赖的学习和记忆是一种行为的任务。它已被广泛应用在神经生物学，神经和神经紊乱的啮齿动物模型的研究。

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

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 ...

科研

神经科学

Design and Construction of a Cost Effective Headstage for Simultaneous Neural Stimulation and Recording in the Water Maze

Headstage preamplifiers and source followers are commonly used to study neural activity in behavioral neurophysiology experiments. Available commercial products are often expensive, not easily customized, and not submersible. Here we describe a method to design and build a customized, integrated circuit headstage for simultaneous 4-channel neural recording and 2-channel simulation in awake, behaving animals. The headstage is designed using a free, commercially available CAD-type design package, and can be modified easily to accommodate different scales (e.g. to add channels). A customized printed circuit board is built using surface mount resistors, capacitors and operational amplifiers to construct the unity gain source follower circuit. The headstage is made water-proof with a combination of epoxy, parafilm and a synthetic rubber putty. We have successfully used this device to record local field potentials and stimulate different brain regions simultaneously via independent channels in rats swimming in a water maze. The total cost is &lt; $30/unit and can be manufactured readily.

We present a low-cost method to design and construct a light headstage pre-amplifier system with simultaneous neural recording and stimulation capability. This device can be waterproofed for use in swimming animals.

同时神经刺激，并记录在水迷宫的设计和构建一个具有成本效益探头

Headstage preamplifiers and source followers are commonly used to study neural activity in behavioral neurophysiology experiments. Available commercial ...

C. elegans Positive Butanone Learning, Short-term, and Long-term Associative Memory Assays

The memory of experiences and learned information is critical for organisms to make choices that aid their survival. C. elegans navigates its environment through neuron-specific detection of food and chemical odors1, 2, and can associate nutritive states with chemical odors3, temperature4, and the pathogenicity of a food source5.
Here, we describe assays of C. elegans associative learning and short- and long-term associative memory. We modified an aversive olfactory learning paradigm6 to instead produce a positive response; the assay involves starving ~400 worms, then feeding the worms in the presence of the AWC neuron-sensed volatile chemoattractant butanone at a concentration that elicits a low chemotactic index (similar to Toroyama et al.7). A standard population chemotaxis assay1 tests the worms' attraction to the odorant immediately or minutes to hours after conditioning.
After conditioning, wild-type animals' chemotaxis to butanone increases ~0.6 Chemotaxis Index units, its "Learning Index". Associative learning is dependent on the presence of both food and butanone during training. Pairing food and butanone for a single conditioning period ("massed training") produces short-term associative memory that lasts ~2 hours. Multiple conditioning periods with rest periods between ("spaced training") yields long-term associative memory (&lt;40 hours), and is dependent on the cAMP Response Element Binding protein (CREB),6 a transcription factor required for long-term memory across species.8
Our protocol also includes image analysis methods for quick and accurate determination of chemotaxis indices. High-contrast images of animals on chemotaxis assay plates are captured and analyzed by worm counting software in MatLab. The software corrects for uneven background using a morphological tophat transformation.9 Otsu's method is then used to determine a threshold to separate worms from the background.10 Very small particles are removed automatically and larger non-worm regions (plate edges or agar punches) are removed by manual selection. The software then estimates the size of single worm by ignoring regions that are above a specified maximum size and taking the median size of the remaining regions. The number of worms is then estimated by dividing the total area identified as occupied by worms by the estimated size of a single worm.
We have found that learning and short- and long-term memory can be distinguished, and that these processes share similar key molecules with higher organisms.6,8 Our assays can quickly test novel candidate genes or molecules that affect learning and short- or long-term memory in C. elegans that are relevant across species.

C. elegans Positive Butanone Learning, Short-term, and Long-term Associative Memory Assays

Here we describe methods to test C. elegans associative learning and short- and long-term associative memory. These population assays employ the worms abilities to chemotax toward volatile odorants, and form positive associations upon pairing food with the chemoattractant butanone. Increasing the number of conditioning periods induces long-term memory.

线虫的积极丁酮学习，短期，长期联想记忆检测

The memory of experiences and learned information is critical for organisms to make choices that aid their survival. C. elegans navigates its environment ...

Assaying Locomotor, Learning, and Memory Deficits in Drosophila Models of Neurodegeneration

Advances in genetic methods have enabled the study of genes involved in human neurodegenerative diseases using Drosophila as a model system1. Most of these diseases, including Alzheimer's, Parkinson's and Huntington's disease are characterized by age-dependent deterioration in learning and memory functions and movement coordination2. Here we use behavioral assays, including the negative geotaxis assay3 and the aversive phototaxic suppression assay (APS assay)4,5, to show that some of the behavior characteristics associated with human neurodegeneration can be recapitulated in flies. In the negative geotaxis assay, the natural tendency of flies to move against gravity when agitated is utilized to study genes or conditions that may hinder locomotor capacities. In the APS assay, the learning and memory functions are tested in positively-phototactic flies trained to associate light with aversive bitter taste and hence avoid this otherwise natural tendency to move toward light. Testing these trained flies 6 hours post-training is used to assess memory functions. Using these assays, the contribution of any genetic or environmental factors toward developing neurodegeneration can be easily studied in flies.

Assaying Locomotor, Learning, and Memory Deficits in Drosophila Models of Neurodegeneration

Behavior assays for measuring locomotor functions, learning, and memory abilities in Drosophila.

含量的运动能力，学习和记忆的赤字果蝇</em神经退行性疾病>模型

Advances in genetic methods have enabled the study of genes involved in human neurodegenerative diseases using Drosophila as a model system1. Most of ...

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 ...

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

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

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

阈上的神经活动的单细胞和单穗的分辨率的光学的方法记录

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

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

The ability to probe defined neural circuits in awake, freely-moving animals with cell-type specificity, spatial precision, and high temporal resolution has been a long sought tool for neuroscientists in the systems-level search for the neural circuitry governing complex behavioral states. Optogenetics is a cutting-edge tool that is revolutionizing the field of neuroscience and represents one of the first systematic approaches to enable causal testing regarding the relation between neural signaling events and behavior. By combining optical and genetic approaches, neural signaling can be bi-directionally controlled through expression of light-sensitive ion channels (opsins) in mammalian cells. The current protocol describes delivery of specific wavelengths of light to opsin-expressing cells in deep brain structures of awake, freely-moving rodents for neural circuit modulation. Theoretical principles of light transmission as an experimental consideration are discussed in the context of performing in vivo optogenetic stimulation. The protocol details the design and construction of both simple and complex laser configurations and describes tethering strategies to permit simultaneous stimulation of multiple animals for high-throughput behavioral testing.

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

Optogenetics has become a powerful tool for use in behavioral neuroscience experiments. This protocol offers a step-by-step guide to the design and set-up of laser systems, and provides a full protocol for carrying out multiple and simultaneous in vivo optogenetic stimulations compatible with most rodent behavioral testing paradigms.

在啮齿动物中枢神经系统的体内光遗传学刺激

The ability to probe defined neural circuits in awake, freely-moving animals with cell-type specificity, spatial precision, and high temporal resolution ...

A Lateralized Odor Learning Model in Neonatal Rats for Dissecting Neural Circuitry Underpinning Memory Formation

Rat pups during a critical postnatal period (&#8804; 10 days) readily form a preference for an odor that is associated with stimuli mimicking maternal care. Such a preference memory can last from hours, to days, even life-long, depending on training parameters. Early odor preference learning provides us with a model in which the critical changes for a natural form of learning occur in the olfactory circuitry. An additional feature that makes it a powerful tool for the analysis of memory processes is that early odor preference learning can be lateralized via single naris occlusion within the critical period. This is due to the lack of mature anterior commissural connections of the olfactory hemispheres at this early age. This work outlines behavioral protocols for lateralized odor learning using nose plugs. Acute, reversible naris occlusion minimizes tissue and neuronal damages associated with long-term occlusion and more aggressive methods such as cauterization. The lateralized odor learning model permits within-animal comparison, therefore greatly reducing variance compared to between-animal designs. This method has been used successfully to probe the circuit changes in the olfactory system produced by training. Future directions include exploring molecular underpinnings of odor memory using this lateralized learning model; and correlating physiological change with memory strength and durations.

This protocol introduces lateralized early odor preference learning in rats using acute single naris occlusion. Lateralized learning permits the examination of behavioral outcomes and underpinning biological mechanisms within the same animals, reducing variance induced by between-animal designs. This protocol can be used to investigate molecular mechanisms underpinning early odor learning.

新生大鼠于A气味偏侧学习模型解剖神经回路托换记忆的形成

Rat pups during a critical postnatal period (&#8804; 10 days) readily form a preference for an odor that is associated with stimuli mimicking maternal ...

Intracerebroventricular Injection of Amyloid-&#946; Peptides in Normal Mice to Acutely Induce Alzheimer-like Cognitive Deficits

Amyloid-&#946; (A&#946;) is a major pathological mediator of both familial and sporadic Alzheimer's disease (AD). In the brains of AD patients, progressive accumulation of A&#946; oligomers and plaques is observed. Such A&#946; abnormalities are believed to block long-term potentiation, impair synaptic function, and induce cognitive deficits. Clinical and experimental evidences have revealed that the acute increase of A&#946; levels in the brain allows development of Alzheimer-like phenotypes. Hence, a detailed protocol describing how to acutely generate an AD mouse model via the intracerebroventricular (ICV) injection of A&#946; is necessary in many cases. In this protocol, the steps of the experiment with an A&#946;-injected mouse are included, from the preparation of peptides to the testing of behavioral abnormalities. The process of preparing the tools and animal subjects before the injection, of injecting the A&#946; into the mouse brain via ICV injection, and of assessing the degree of cognitive impairment are easily explained throughout the protocol, with an emphasis on tips for effective ICV injection of A&#946;. By mimicking certain aspects of AD with a designated injection of A&#946;, researchers can bypass the aging process and focus on the downstream pathology of A&#946; abnormalities.

Intracerebroventricular Injection of Amyloid-&amp;#946; Peptides in Normal Mice to Acutely Induce Alzheimer-like Cognitive Deficits

The amyloid-&#946; (A&#946;)-injected animal model enables the administration of a defined quantity and species of A&#946; fragments and reduces individual differences within each study group. This protocol describes the intracerebroventricular (ICV) injection of A&#946; without stereotactic instruments, enabling the production of Alzheimer-like behavioral abnormalities in normal mice.

正常小鼠β淀粉样肽侧脑室注射敏锐地诱导老年痴呆样认知缺陷

Amyloid-&#946; (A&#946;) is a major pathological mediator of both familial and sporadic Alzheimer's disease (AD). In the brains of AD patients, ...

Assessing Spatial Learning and Memory in Small Squamate Reptiles

Clinical research has leveraged a variety of paradigms to assess cognitive decline, commonly targeting spatial learning and memory abilities. However, interest in the cognitive processes of nonmodel species, typically within an ecological context, has also become an emerging field of study. In particular, interest in the cognitive processes in reptiles is growing although experimental studies on reptilian cognition are sparse. The few reptilian studies that have experimentally tested for spatial learning and memory have used rodent paradigms modified for use in reptiles. However, ecologically important aspects of the physiology and behavior of this taxonomic group must be taken into account when testing for spatially based cognition. Here, we describe modifications of the dry land Barnes maze and associated testing protocol that can improve performance when probing for spatial learning and memory ability in small squamate reptiles. The described paradigm and procedures were successfully used with male side-blotched lizards (Uta stansburiana), demonstrating that spatial learning and memory can be assessed in this taxonomic group with an ecologically relevant apparatus and protocol.

This paper describes a modification of the Barnes maze, a standard rodent paradigm used to assess spatial memory and learning, for use in small squamate reptiles.

评估空间学习记忆小鳞类爬行动物

Clinical research has leveraged a variety of paradigms to assess cognitive decline, commonly targeting spatial learning and memory abilities. However, ...