撰文人先前发表的工作得到了国家精神健康研究所 (MH088480) 的资助。作者想感谢她的前任导师 Dr. 在该项目中对他的支持。这份出版物得到了国家卫生研究院 (T32 DA007135) 的资助。

在这里执行的所有程序都是由动物保育和使用委员会提交并批准的, 并按照 NIH 指南进行. 
 1. 对象选择和实验设置 <ol> <li> 选择不同的对象, 它们很容易被小鼠区分, 但具有类似的复杂程度 (纹理、形状、颜色图案和亮度, 等 )为了最小化可能导致结果偏差的任何潜在的对象偏好 (请参见 Ennaceur 2010 以了解对象选择 7 的全面描述). 
	<ol> <li> 测试先天偏好和歧视 (参见步骤2和 #38; 3). </li>
		<li> 使用 mouse-sized 或仅略大的对象来鼓励探测 ( 图 1 )。为了减少诱导性偏好, 确保小鼠能够在两个物体上攀爬, 或者既不在物体上攀爬, 虽然爬上一个物体的能力可能会增加勘探的兴趣 (只是坐在一个物体上的时间并不算在勘探时间内). </li>
		<li> 每个对象至少有2份副本。然而, 为了尽量减少潜在的气味提示在测试期间, 有至少3每个对象, 重复为训练和一个为测试. </li>
		<li> 使用由 non-breakable 材料制成的物体, 由于试验过程中的损坏而造成的物体损失可能会干扰测试的连续性, 并可能对动物造成伤害或伤害. </li>
		<li> 在使用之前和之间彻底清理对象 (70% 卷/卷的乙醇是适当的) </li>
	</ol> </li> <li> 以最小化明亮照明的应力, 使用漫反射、低光照, 与迷宫的中心照明约20勒克斯。使用类似于正常住房条件的温度和湿度。如果老鼠被从一个房间搬到另一个房间进行实验, 那么在每天使用之前, 要让老鼠适应新房间至少1小时. </li>
	<li> 在训练前处理好小鼠。因为测试依赖于啮齿目动物的自然倾向探索新奇, 减少任何压力或焦虑从处理可能干扰他们的欲望探索竞技场, 并且随后, 对象。理想情况下, 在测试 2 , 15 之前, 处理鼠标 1-2 次/天至少1分钟, 1 到29周。
	<li> 对于主竞技场, 使用方形腔 (大约40厘米 x 40 cm x 40 厘米) 制成的白色或黑色, 无孔塑料, 对比的颜色的鼠标。或者, 使用一个圆形竞技场 ed. 具体地说, 当使用焦虑小鼠, 可能会坐在一个正方形竞技场的角落, 一个圆形竞技场可能会更好地鼓励探索行为 18 , 30 . 
	<ol> <li> 在训练日, 将2相同的对象放在对角线上 ( 即: 一个在西北角, 一个在 SE 角)。如果物体太轻, 可以被老鼠移动, 请把它们固定在地板上。可拆卸安装腻子工作良好. </li>
		<li> 对于测试日, 请使用每个对象的 3 rd 副本将一个熟悉的对象和一个新对象放置在与训练日相同的对角线上 ( 图 2 ). </li>
		<li> 平衡每组对象的使用, 使每个对象作为一个熟悉的对象和新对象被同等地使用。此外, 平衡的位置, 小说对象的每一个竞技场的4个角落。一定要注意哪种对角线用于哪种动物, 以便训练日使用的对角线与在测试日使用的每个鼠标相同. </li>
		<li> 彻底清洁设备和物件, 以消除气味提示之前和之间使用 (70% 卷/卷乙醇是适当的). </li>
	</ol> </li> <li> 将照相机直接置于设备的上方, 以获得最佳的探测视图。仅使用带有鼻点检测功能的软件进行精确分析。如果使用软件, 请在开始之前将背景图像与所处的对象进行捕获。在软件中, 将勘探区域设置为大约 2-3 厘米左右的对象. </li>
	<li> 以减少实验者的干扰, 记录试验并在以后进行评分。使射手对实验条件视而不见。如果在实验中手动评分, 请确保试验者从竞技场上超过1米, 而对鼠标不可见. 
	注意: 任何时候, 鼠标在对象上坐在没有主动触须扫或嗅探不算作勘探时间. </li>
	<li> 将探测定义为当鼠标和 #39 的噪音指向对象, 并在对象的 2-3 厘米内, 用主动触须清扫或嗅探。不要指望任何时间坐在物体上, 没有积极探索的迹象. </li>
</ol> 2。必要的试验性试验 <ol> <li> 测试诱导偏好 <ol> 用于习惯化, 将鼠标从其主笼中取出, 并将其放在开放竞技场的中间。允许鼠标自由浏览5分钟. </li>
		<li> 在训练日 (T1) 上, 将2不同的对象放置在设备的相对象限中 ( 即 、NW 和 SE 角)。将鼠标从其主笼子中取出, 并将其放在开放竞技场的中央。允许自由探测10分钟 </li>
		<li> 计算判别指数。如果没有诱导偏好, 则歧视指数应为或接近于零。任何显示首选项的对象都不应用于对. </li>
	</ol> </li> <li> 对辨别能力的测试 <ol> 为适应, 从它的主笼中移除鼠标并将其放置在开放竞技场的中央。允许鼠标自由浏览5分钟. </li>
		<li> 在训练会话 (T1) 上, 在相反的象限中放置2相同的对象。将鼠标放在竞技场的中心, 允许鼠标探索10分钟. </li>
		<li> 在测试会话 (T2) 上, 将 T1 中使用的对象之一和在相反象限中的一个新对象 (即 NW 和 SE 角) 放在一起。六十分钟后 T1, 将鼠标放在竞技场的中心, 允许自由探索10分钟. </li>
		<li> 计算判别指数。在60分钟的保留间隔, 歧视指数应高于 0.25. </li>
	</ol> </li> </ol> 3。实验过程 <ol> <li> 自适应 <ol> 将鼠标从其主笼中移除, 并将其放在打开的空竞技场中间。允许自由探索竞技场5分钟。一旦家庭笼是空的, 保存它作为一个保持笼的第二天. </li>
		<li> 在5分钟的末尾, 将鼠标移出并放置在一个固定的保持架中。不要将鼠标返回到原来的笼子, 否则可能会影响待测试的小鼠的行为. </li>
		<li> 使用70% 卷/卷乙醇彻底清洁小鼠之间的设备. 
		注意: 在习惯性中, 焦虑样的行为可以通过计算中心的时间来评估 (见普鲁特河和 #38; Belzung 2003 31 )。在考虑 T1 的时间长短时, 这是一个有用的度量。高焦虑小鼠可能需要10分钟的会议才能达到最低勘探标准. </li>
	</ol> </li> <li> 训练 (T1) <ol> <li> 将两个相同的对象置于竞技场的相反象限 ( 即 、NE 角和 SW 角). </li>
		<li> 自适应后24小时, 将鼠标从其主笼中取出, 并将其置于竞技场的中心, 与2相同的对象相等. </li>
		<li> 允许自由探测至少5分钟。如果使用一株已知具有低运动或探索活性的小鼠, ( 即 , 大多数小鼠在二十年代对两个物体的探索至少达不到5分钟, 如在实验或文献中所指出的那样), 将试验扩展到所有麦克风的10分钟队列中的 e. </li>
		<li> 在试验结束时, 取出鼠标并将其放在保持架中。一旦主笼为空, 将其保存以供使用作为在测试日的控股笼. </li>
		<li> 使用70% 卷/卷乙醇彻底清洁小鼠之间的器具和物体. </li>
	</ol> </li> <li> 测试 (T2) <ol> <li> 在 T1 ( 即 , 熟悉的对象) 中使用一个对象, 在竞技场的相反象限中放置一个新对象。使用在 T1 期间为每个鼠标所使用的相同位置. </li>
		<li> 在 T1 到 T2 间隔选择时, 将鼠标从其主笼中移除, 并将其置于中心舞台上, 与熟悉的对象和新对象相等. 
		注意: 在保留间隔24小时, 大多数小鼠将无法区分熟悉和新奇的对象 (通常是-0.2 和 #60; d2 和 #60; 0.2, 与正控制 32 相比)。如果测试促效果, 请使用此时间点来探测内存增强。要探测内存不足, 请使用较短的保留间隔, 在20分钟到4小时之间, 这取决于鼠标的应变. </li>
		<li> 允许自由探测10分钟。在试验结束时, 取出鼠标并将其放置在保持架中. </li>
	</ol> </li> <li> 对于 T1 和 T2, 评分前5分钟。如果这两个物体的最小勘探时间都不符合二十年代, 则继续得分超过5分钟, 直到总勘探超过二十年代. </li>
</ol> 4。数据分析 <ol> <li> 排除条件 <ol> 在培训 (T1) 和测试 (T2) 期间, 计算每个会话的两个对象的总探测时间 (e1 和 e2)。大多数小鼠应达到最小勘探总量为两个对象的二十年代5分钟. </li>
		<li> 将 T1 和 T2 时间延长到10分钟, 对于低勘探的小鼠, 不符合这一最低标准5分钟, 在试点试验中观察到. </li>
		<li> 评分行为为5分钟或超过5分钟, 直到他们达到二十年代的最低标准. </li>
		<li> 如果小鼠在10分钟内没有达到对 T1 或 T2 两个物体的勘探最小值, 则排除在分析之外, 因为它无法证实他们花了足够的时间去探索学习/辨别. </li>
	</ol> </li> <li> 绝对 vs 相对分析 注: 分析的不同公式及其彼此之间的关系可以在 表 1 中看到。
	<ol> <li> 将 e1 计算为2相同对象的训练期间的总探测时间, 其中 a1 和 a2 是相同的对象. 
		 e1 = a1 + a2 </li>
		<li> 将 e2 计算为熟悉对象 (a) 和新对象 (b) 测试期间的总探测时间. 
		 e2 = a + b </li>
		<li> 计算 d1 只是花在探索新对象上的时间减去探索熟悉对象所花费的时间。绝对歧视措施 (d1) 没有考虑到小鼠或治疗组在探索时间上的差异, 虽然在某些情况下, 它可能是一个更敏感的措施 2 . 
		 d1 = b-a </li>
		<li> 计算 d2 作为探索新对象所花费的时间减去探索熟悉对象所花费的时间除以总探测时间。最常用的措施是一个相对的歧视的价值通常被称为歧视指数 (d2), 这是不受不同的勘探时间的差异。这意味着所有值将介于-1 和 + 1 之间. 
		 d2 = d1/e2 </li>
		<li> 或者, 计算识别或首选项索引 (d3) 3 。这是花时间探索新对象除以总时间。这意味着所有值将介于0和1之间。它通常乘以 100, 并用作百分比值. 
		 d3 = b/e2*100 </li>
	</ol> </li> <li> 统计分析 <ol> <li> 使用每个组的平均判别值, 使用 one-way 方差计算确定内存性能。要进一步分析, 请与处理过的 vs 车辆条件和正/负控制组进行 two-way 临时 比较. </li>
	</ol>
	</li>
</ol>

<ol>
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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+anatomy,+physiology+and+functions+of+the+perirhinal+cortex.">The anatomy, physiology and functions of the perirhinal cortex.</a> Curr. Opin. Neurobiol. 6, 179-186 (1996).</li><li>Wan, H., Aggleton, J. P., Brown, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Different+contributions+of+the+hippocampus+and+perirhinal+cortex+to+recognition+memory.">Different contributions of the hippocampus and perirhinal cortex to recognition memory.</a> J. Neurosci. 19 (3), 1142-1148 (1999).</li><li>Warburton, E. C., Brown, M. 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作者没有什么要透露的。

对小鼠学习记忆的研究是一种有效、灵活的方法。在设置实验时, 重要的是要考虑一些可能影响结果的变量。正如在代表性的结果中所讨论的, 小鼠的应变会影响勘探时间和保留间隔。勘探时间的减少可能歪曲或掩盖结果在绝对歧视分析2,3,5,30,32。某些品系的小鼠可能有较低?...

对象识别测试 (也称为新的对象识别测试) 是一种相对快速、高效的测试小鼠学习记忆不同阶段的方法。它最初被描述了 Ennaceur 并且德拉库尔在1988年和主要使用在大鼠1;然而, 自那时以来, 它已经成功地适应了使用在小鼠2,3,4,5,6,7。测试仅依赖于三会话: 一个适应会话、一个培训会话和一个测试会话。训练只涉及对两个相同对象的可视化探索, 而测试会话则需要用一个新的对象替换以前探索的对象之一。因为啮齿目动物天生偏爱新奇, 记忆熟悉对象的啮齿目动物将花费更多时间探索新的对象7,8,9。
对其他啮齿动物记忆测试的主要好处是, 它依赖于啮齿动物的自然倾向, 探索新奇的8。因此, 不需要大量的训练课程或任何积极或消极的强化来激励行为。这意味着, 相对于其他测试10、11、12、13、14、15, 它的压力大大降低, 并且需要显著运行时间比其他常用的内存测试更少, 如莫里斯水迷宫或巴恩斯迷宫, 两者都可能长达一周或更长。因此, 该方法的条件更接近于那些用于研究人类认知, 增加了对许多其他啮齿动物记忆测试的生态有效性的测试。同样, 因为它是一个简单的视觉召回任务, 它已经成功地适应了许多物种, 包括人类和非人灵长类的使用, 以评估声明性内存的不同间方面2,16 ,17。最后, 可以很容易地修改, 以检查不同阶段的学习和记忆 (即, 获取, 整合, 或召回), 以评估不同类型的内存 (如如, 空间内存), 或评估不同的保留间隔 (即, 短期vs长期内存)。
它的通用性为无数的研究应用提供了一个平台。研究可以利用药理学的药物来扰乱或增强记忆力。改变药物管理的时间在训练之前或之后, 或在测试之前可以提示在潜在的神经机制, 导致中断或增强内存6,18,19, 20. 以类似的方式, optogenetic 技术可以在这些相同的不同时间点上使用, 以观察有助于学习和记忆不同阶段的神经激活/抑制。它也适用于评估转基因动物、病变研究或神经退行性模型或衰老研究中的差异21,22,23,24,25,26,27,28. 训练和测试之间的时间 (称为保留间隔) 可以更改为对短期和长期内存26中的任何这些更改进行评估。最终, 该药物可以被用来作为一个工具, 研究药理, 遗传, 和神经学的变化, 以学习和记忆, 或这些工具可以用来研究的基础, 学习和记忆的支撑。

<table><tbody><tr><td>Open Field Box</td><td>Panlab/Harvard Apparatus</td><td>LE800SC</td><td>Available in grey, white, or black</td></tr><tr><td>ANY-maze</td><td>Stoelting Co.</td><td>60000</td><td>Behavior tracking system</td></tr><tr><td>EthoVisionXT 12</td><td>Noldus</td><td></td><td>Behavior tracking system; requires 3 point tracking</td></tr><tr><td>Video Camera</td><td>Any</td><td></td><td>Video camera should be mounted directly overhead of the apparatus</td></tr><tr><td>70% Ethanol&amp;nbsp;</td><td>Fisher Scientific</td><td>BP2818-4</td><td>Prior to starting testing and in between trials, each object should be carefully cleaned. The floor and walls of the apparatus should also be cleaned.&amp;nbsp;</td></tr></tbody></table>

novel object recognition test for the investigation of learning and memory in mice

物体识别试验是对小鼠学习记忆各方面进行研究的一种常用的行为检测方法。该运动是相当简单, 可以完成超过3天: 习惯日, 训练日, 和测试日。在训练过程中, 允许鼠标探索2相同的对象。在测试日, 一个训练对象被一个新的对象替换。因为老鼠天生偏爱新奇, 如果老鼠认出了熟悉的物体, 它就会把大部分时间花在新奇的物体上。由于这种先天的偏好, 不需要积极或消极的强化或长期的训练计划。此外, 还可以对许多应用程序进行修改。保留间隔可以缩短以检查短期内存, 或延长以探测长期内存。药理干预可以在训练前、训练后的不同时间使用, 也可在召回前研究不同的学习阶段 (即、习得、早或晚合并或召回)。总的来说, 这是一个相对低, 有效的测试记忆的小鼠, 并适合于检测神经心理学的变化后, 药理, 生物, 或基因的操纵。

在图 2中显示了一个通用的实验性设置。在习惯日 (T0) 老鼠被安置在空的竞技场为 5 min. 二十四小时后, 老鼠被放回房间里, 有2相同的物体, 允许自由探索多达10分钟 (T1)。在测试日 (T2), 老鼠再被安置在竞技场, 但与一个熟悉的对象和一个新颖的对象, 并且允许探索为 10 min。保留间隔, T1 和 T2 之间的时间, 可以改变, 这取决于实验的最终目标。在有代表性...

Watch this Scientific Journal Video about Novel Object Recognition Test for the Investigation of Learning and Memory in Mice at JoVE.com

Novel Object Recognition Test for the Investigation of Learning and Memory in Mice

目标识别试验是评价小鼠学习记忆的一种简单有效的方法。方法如下所述。

小鼠学习记忆研究的新对象识别实验

The object recognition test (ORT) is a commonly used behavioral assay for the investigation of various aspects of learning and memory in mice. The ORT is ...

novel-object-recognition-test-for-investigation-learning-memory

Research

JoVE Journal

Behavior

72.5K 次观看.  Icahn School of Medicine at Mount Sinai. 目标识别试验是评价小鼠学习记忆的一种简单有效的方法。方法如下所述。

视频: Novel Object Recognition Test for the Investigation of Learning and Memory in Mice

Novel Object Exploration as a Potential Assay for Higher Order Repetitive Behaviors in Mice

Restricted, repetitive behaviors (RRBs) are a core feature of autism spectrum disorder (ASD) and disrupt the lives of affected individuals. RRBs are commonly split into lower-order and higher-order components, with lower order RRBs consisting of motor stereotypies and higher order RRBs consisting of perseverative and sequencing behaviors. Higher order RRBs are challenging to model in mice. Current assays for RRBs in mice focus primarily on the lower order components, making basic biomedical research into potential treatments or interventions for higher-order RRBs difficult. Here we describe a new assay, novel object exploration. This assay uses a basic open-field arena with four novel objects placed around the perimeter. The test mouse is allowed to freely explore the arena and the order in which the mouse investigates the novel objects is recorded. From these data, patterned sequences of exploration can be identified, as can the most preferred object for each mouse. The representative data shared here and past results using the novel object exploration assay illustrate that inbred mouse strains do demonstrate different behavior in this assay and that strains with elevated lower order RRBs also show elevated patterned behavior. As such, the novel object exploration assay appears to possess good face validity for higher order RRBs in humans and may be a valuable assay for future studies investigating novel therapeutics for ASD.

Higher order restricted, repetitive behaviors (RRBs) disrupt the lives of affected individuals. These behaviors are challenging to model in rodents, making basic biomedical research into potential treatments or interventions for RRBs difficult. Here we describe novel object exploration as a potential assay for higher order RRBs in mice.

新对象的探索在小鼠电位测定高阶重复的行为

Restricted, repetitive behaviors (RRBs) are a core feature of autism spectrum disorder (ASD) and disrupt the lives of affected individuals.  RRBs are ...

科研

行为学

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 ...

神经科学

Novel Object Recognition and Object Location Behavioral Testing in Mice on a Budget

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various physiological or pathological changes. The object location task (OLT) and the novel object recognition task (NORT) are two effective behavioral tasks commonly used to reveal the function and relative health of specific brain regions involved in memory. While both of these tests exploit the inherent preference of mice for the novelty to reveal memory for previously encountered objects, the OLT primarily evaluates spatial learning, which relies heavily on hippocampal activity. The NORT, in contrast, evaluates non-spatial learning of object identity, which relies on multiple brain regions. Both tasks require an open-field-testing arena, objects with equivalent intrinsic value to mice, appropriate environmental cues, and video recording equipment and the software. Commercially available systems, while convenient, can be costly. This manuscript details a simple, cost-effective method for building the arenas and setting up the equipment necessary to perform the OLT and NORT. Furthermore, the manuscript describes an efficient testing protocol that incorporates both OLT and NORT and provides typical methods for data acquisition and analysis, as well as representative results. Successful completion of these tests can provide valuable insight into the memory function of various mouse model systems and appraise the underlying neural regions that support these functions.

Here we provide a protocol which includes comprehensive instructions for the economical establishment of murine object location and novel object recognition behavioral testing, including the design, cost, and construction of required equipment as well as execution of behavioral testing, data collection, and analysis.

一种新的小鼠目标识别与目标定位行为测试

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various ...

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 ...

Assessment of Memory Function in Pilocarpine-induced Epileptic Mice

Cognitive impairment is one of the most common comorbidities in temporal lobe epilepsy. To recapitulate epilepsy-associated cognitive decline in an animal model of epilepsy, we generated pilocarpine-treated chronic epileptic mice. We present a protocol for three different behavioral tests using these epileptic mice: novel object location (NL), novel object recognition (NO), and pattern separation (PS) tests to evaluate learning and memory for places, objects, and contexts, respectively. We explain how to set the behavioral apparatus and provide experimental procedures for the NL, NO, and PS tests following an open field test that measures the animals&#8217; basal locomotor activities. We also describe the technical advantages of the NL, NO, and PS tests with respect to other behavioral tests for assessing memory function in epileptic mice. Finally, we discuss possible causes and solutions for epileptic mice failing to make 30 s of good contact with the objects during the familiarization sessions, which is a critical step for successful memory tests. Thus, this protocol provides detailed information about how to assess epilepsy-associated memory impairments using mice. The NL, NO, and PS tests are simple, efficient assays that are appropriate for the evaluation of different kinds of memory in epileptic mice.

This article presents experimental procedures for assessing memory impairments in pilocarpine-induced epileptic mice. This protocol can be used to study the pathophysiologic mechanisms of epilepsy-associated cognitive decline, which is one of the most common comorbidities in epilepsy.

皮洛卡平诱导癫痫小鼠记忆功能评估

Cognitive impairment is one of the most common comorbidities in temporal lobe epilepsy. To recapitulate epilepsy-associated cognitive decline in an animal ...

Simultaneous Monitoring of Wireless Electrophysiology and Memory Behavioral Test as a Tool to Study Hippocampal Neurogenesis

Brainwaves amplitude obtained from electroencephalography (EEG) has been well-recognized as a basis for cognitive capacity, memory, and learning on animals and humans. Adult neurogenesis mechanism is also linked to memory and learning improvement. Traditionally, researchers used to assess learning and memory parameters in rodent models by behavioral tasks. Therefore, the simultaneous monitoring of behavioral changes and EEG is particularly interesting in correlating data between brain activity and task-related behaviors. However, most of the equipment required to perform both studies are either complex, expensive, or uses a wired setup network that hinders the natural animals&#8217; movement. In this study, EEG was recorded with a wireless electrophysiology device along with the execution of a novel object recognition task (NORT). The animal&#8217;s behavior was monitored simultaneously by a video tracking system. Both recordings were analyzed offline by their timestamps which were synched to link EEG signals with the animal&#8217;s actions. Subjects consist of adult Wistar rats after medium-term environmental enrichment treatment. Six skull screw electrodes were fixed in pairs on both hemispheres over frontal, central, and parietal regions and were referenced to an electrode located posterior of the nasal bone. NORT protocol consists of exposing the animal to two identical objects for 10 min. After 2 h and 24 h, one of the objects was replaced with a novel one. Exploration time for each object was monitored by a behavioral tracking software (BTS) and EEG data recording. The analysis of the EEG synced with behavioral data consists of estimations of alpha and beta relative band power and comparisons between novel object recognition versus familiar object exploration, between three experimental stages. In this manuscript, we have discussed electrodes manufacturing process, epidural electrodes implantation surgery, environmental enrichment protocol, NORT protocol, BTS setup, EEG &#8211; BTS coupling for simultaneous monitoring in real-time, and EEG data analysis based on automatic events detection.

The protocol presented here provides information on the simultaneous electroencephalography (EEG) and behavioral assessment in real-time. We have discussed all steps involved in this protocol as an attractive solution for researchers in many fields of neuroscience, particularly in learning and memory areas.

同时监测无线电生理学和记忆行为测试作为研究海马神经发生的工具

Brainwaves amplitude obtained from electroencephalography (EEG) has been well-recognized as a basis for cognitive capacity, memory, and learning on ...

A Within-Subject Experimental Design using an Object Location Task in Rats

Object place recognition is a prominent method used to investigate spatial memory in rodents. This object place recognition memory forms the basis of the object location task. This paper provides an extensive protocol to guide the establishment of an object location task with the option of up to four repetitions using the same cohort of rats. Both weak and strong encoding protocols can be used to study short- and long-term spatial memories of varying strength and to enable the implementation of relevant memory-inhibiting or -enhancing manipulations. In addition, repetition of the test with the counterbalancing presented here allows the combination of results from two or more tests for within-subject comparison to reduce variability between rats. This method helps to increase statistical power and is strongly recommended, particularly when running experiments that produce high variation in individual behavior. This directly refines the study by increasing the data obtained from each animal and reducing the overall number of animals needed.&#160;Finally, implementation of the repeated object location task increases the efficiency of studies that involve surgical procedures by saving time and labor.

This protocol provides detailed steps for an object location task with four repetitions using the same cohort of rats. Weak and strong encoding can produce short- and long-term memories. The flexibility of the protocol with repetition can be beneficial for studies involving surgical operations by saving time and labor.

使用大鼠对象定位任务的主体内实验设计

Object place recognition is a prominent method used to investigate spatial memory in rodents. This object place recognition memory forms the basis of the ...

Assessment of Spontaneous Alternation, Novel Object Recognition and Limb Clasping in Transgenic Mouse Models of Amyloid-&#946; and Tau Neuropathology

Here we describe a staged, behavioral testing approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional motor behaviors in transgenic mouse models of &#946;-amyloidosis and tauopathy. The paradigm includes tests for spontaneous alternation in a Y-maze, novel object recognition, and limb clasping. These tests were selected because they: 1) interrogate function of cognitive or motor domains and the correlate neural circuitry relevant to the human disease state, 2) have clearly defined endpoints, 3) have easily implementable quality control checks, 4) can be run in a moderate throughput format, and 5) require little intervention by the investigator. These methods are designed for investigators looking to screen compounds for activity in short-term and working memory tasks, or functional motor behaviors associated with Alzheimer's disease mouse models. The methods described here use behavioral tests that engage a number of different brain regions including hippocampus and various cortical areas. Investigators that desire cognitive tests that specifically assess cognition mediated by a single brain region could use these techniques to supplement other behavioral tests.

Described here is a staged, behavioral screening approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional motor behaviors in transgenic mouse models of &#946;-amyloidosis and tauopathy. These methods are optimized to screen compounds for activity in short-term and working memory tasks.

自发交替评估，淀粉样蛋白β和脑神经病理学转基因小鼠模型中的新物体识别和肢体紧扣

Here we describe a staged, behavioral testing approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional ...

医学

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.