我们感谢约翰·韦尔先生和特里Upshall先生为他们的技术专长和援助的摄影和录像。这项工作是通过操作补助JLV从健康研究的加拿大学院和纽芬兰的研究与发展公司和心脏病及中风的加拿大加拿大合作基金会中风回收催化剂赠款的支持。 RBR是基思·格里菲斯纪念心脏及中风基金会研究生奖学金的获得者。

伦理声明：所有实验由纽芬兰的动物保健伦理委员会纪念大学根据加拿大议会关于动物饲养的指导方针批准。 1.小鼠<ol><li>用成年鼠。在这项研究中，成年雄性FVBN小鼠（n = 10）之间2-4个月的被使用。家鼠在12:12小时反向明暗周期，并提供标准啮齿动物食物和水自由采食 。 </li></ol> 2.材料所需圆柱测试<ol><li>获得一台具有透明的顶部拍摄的气瓶检验。表的尺寸是无关紧要的，顶部必须有机玻璃或玻璃，且必须有足够的空间来放置该台下面的反射镜。这允许鼠标从下面录像。使用下表镜子反射通过汽缸的图像。作为替代方案，使用倒置相机（如果可用）。该表在本协议中使用的尺寸为54 x 56×66.5厘米（wxlxh）与51 x 20英寸顶部（WXL）。 </li><li>得到的反射镜。镜子在本协议中所使用的尺寸为34 x58厘米（WXL）。 </li><li>得到透明/有机玻璃筒的鼠标来执行英寸圆柱体在这个协议中使用的尺寸为17.5厘米高，8.8厘米ID，9.5厘米外径与0.35厘米的壁厚。一个更高的汽缸可能需要更积极的小鼠品系。 </li><li>将气缸在桌面和电影在下面的镜面反射。 </li><li>一个摄影机和三脚架是必需的。记录的视频以大约650KB / s，这是每5分钟气缸视频的大约190MB。确保相机具有缩放功能，以确保在气缸涵盖的整个视野​​。 注：在本协议中使用的摄影机是索尼DCR-SR42，40X光学变焦，2000倍数码变焦，680kpix它采用标清，NTSC隔行扫描视频）。 </li><li>获得软件进行分析 - 媒体PL艾尔视频支持，播放速度调节。在这个协议中使用的媒体播放器是VLC媒体播放器V2.1.2。 </li><li>获得的计算机的操作系统能够运行的媒体播放器和监视器的。 </li><li>电子存储的视频是必需的。视频下载到外部硬盘驱动器或将其复制到DVD光盘进行长期存储。 注：在每部影片190MB，84会议将适合在一个16 GB的SD卡和168会议将适合在32GB的SD卡。由于SD介质的相对便宜，并且在一些小鼠中有多少时间需要完成20后轮的不确定性，建议使用32 GB的卡。在目前的研究中，影片是从相机复制到2TB外置硬盘驱动器，然后再复制到DVD作为备份。 </li></ol>气缸测试3.实验装置<ol><li>紧固在下表镜以45度角的桌面。别连接于桌腿此使用两个支架，以suppor吨反射镜的顶部和底部，分别。托架位置将记录在一个侧视图表（ 图1A）和表（ 图1B）的一个面上视图。 </li><li>放置气缸在桌子的中央。绘制四个等距线所在的气缸坐在桌子上用黑色标记（SHARPIE），使得筒可以解除并返回到相同的位置（ 图2）。绘制它们在透明桌面的下侧允许桌面到动物试验之间进行清洗而不溶解的标记油墨。 </li><li>连接相机和三脚架。将相机对准镜子使画面直接翻翻缸筒。确保气缸的充分内壁是可见的和通畅由基座（ 图3）。见设置的侧上视图，包括相机和镜设置的相对角度，从上方（ 图4A），并从表中的电平，包括一个鼠标饲养（ 图4B）。 </li><li>准备提示卡识别之前拍摄每个鼠标。确保卡通常包括用于每个小鼠的识别号码，时间点的测试（ 例如 ，治疗后3天），并拍摄会议日期。不包括在提示卡上的治疗组，以确保实验者被蒙蔽。 </li><li>电影标准的室内照明条件的光这个水平需要清楚地看到围绕气瓶的鼠标移动。 注：如果拍戏可以在一个红灯照相机可能就足够了黑暗，但是人们会首先需要测试的爪子触摸并拖动是否清晰定量可见。 </li></ol> 4.执行<ol><li>开始拍摄。显示适当的鼠标提示卡在镜头前。 </li><li>拍摄提示卡后，立即放下鼠标到从开顶缸。 </li><li>开始拍摄鼠标。在此期间尽量减少噪音，老鼠可能丢失，如果一震探索的兴趣。 </li><li>观察小鼠后探索缸。捕捉视频，直到鼠标执行最低二十后轮。 注意：当两个前爪失去与地面接触和鼠标站在其后腿，则发生后方。 </li><li>擦拭桌面和汽缸，每个鼠标之间取得适当的清洗液消毒和消除气味鼠标。 </li><li>冰点是当小鼠停止探索，并保持安静地坐在四肢着大约5分钟。如果小鼠冻结20后轮发生之前，就可能有必要恢复试验前从气缸移除它们10-20分钟。如果小鼠不执行20后轮，它们从研究中删除。 注：根据我们的经验，老鼠从未需要被排除由于未能探索缸。 </li></ol>气缸测试5.使用评估爪拖分析<ol><li>播放视频时的0.25倍之间和0.67X正常速度的速度取决于如何快速的鼠标探索气缸。使用媒体播放器，提供了更慢的播放速度。 </li><li>量化爪子触摸的总数。爪子触摸发生时，鼠标后轮（ 图5A），触摸气缸（ 图5B）的一侧，随后与两个爪同时（ 图5C）和平台（ 图5D）卸除。爪可以或可以不具有充分手掌接触汽缸壁，但与汽缸壁部分接触必须发生。通过与气缸壁的一个或两个前爪（无论多么短暂）计数的次数鼠标进行接触而站在其后肢的后期间评估爪子触摸。 注意与汽缸壁的接触只算作一个“爪子触摸”或“爪拖”如果鼠标在后方位置 - 站立它的后肢与两个前爪离台面。如果鼠标停留在3点的姿态 - 既后肢和一个前爪在桌面上，并进入与自由爪子触壁 - 这不算是一个爪子触摸。小鼠可后部并碰触所述汽缸壁具有单个前爪和这个被算作一个爪子触摸。注：鼠标也可后移动时它身体的圆筒周围，使两个以上的联系人。这些触点相符 - 每个左前爪touch和为每个右前爪联系。 </li><li>量化爪拖动的数量。 PAW-拖动行为是正常的爪子接触不同。 <ol><li>如果爪子接触到气缸壁采用了全开放式的手掌（ 图6B），它会慢慢地从墙上掉下去，经常有轻微的震颤。运动与拖靠在汽缸壁或者在内侧或向下的方向，（ 图6C）的数字开始落下远COMPLET前伊利（ 图6D）。鼠标会再登陆四肢着地（ 图6F）之前，其未受影响的爪子（ 图6E）下马。这被认为是一个爪子拖动，并应在理货计数。 </li><li>如果爪不与一个完全开放的手掌接触汽缸壁，它将从气缸壁落下离开之前轻擦气缸壁与它的位数。同样，鼠标可能拖累它的爪子在气缸壁上，但拆卸之前不释放完全。这些都被认为PAW-拖动以及触摸和理货应该算作两个。 </li><li>爪也可沿着汽缸壁拖动而鼠标探讨了圆柱体。在这种情况下，爪将跟随鼠标的躯干的扭转，因为它拆卸（ 图7E）之前探讨左侧的原来的位置（ 图7A-D）或右。这不被认为是一个爪拖动的，因为它依赖于小鼠随机选择一个方向探索和不依赖于它的皮质半球受损。 </li></ol></li><li>每个爪子触摸总数在会话期间PAW-拖动表示为百分比爪一拖再拖。表达爪拖动为每个单独前肢爪总接触的百分比数。 </li><li>导致一爪子拖动触摸算作一个爪子拖动，同时触摸。因此，如果一个鼠标每次拖动它的爪子其爪子所述汽缸壁接触，爪拖动百分比表示为100％。 </li></ol> 6.附加的实验设计建议<ol><li>为了尽量减少多余的变量： <ol><li>测试小鼠在同一时间上的每个测试日。在其唤醒周期测试小鼠。小鼠保持在12小时反光照周期有利于性能。 </li><li>小鼠可能不愿意探索如果因噪音或小说环境强调圆柱体。在他们的动物收容室或一室老鼠的测试他们bEEN熟悉减轻压力。这可能发生，如果房间吵，如果鼠标已经推挤进入气缸之前，或由于习惯。 注意：测试应一次实验操作前进行以用作基线读数。操纵后，测试天都在实验者的判断，尽管它是应避免过度的短时间内过多暴露到气缸。 </li><li>老鼠可能会变得不愿意后6-7后暴露于气缸。对于目前的研究中，将小鼠在汽缸中总共七次，缺血前和天1，3，7，14，21和28后的手术进行测试。 注：在本研究中，我们使用了FVBN小鼠品系。我们先前测试之C57B1 / 6小鼠的气缸和观察到的爪拖动行为以下内容的ET-1的缺血性损伤的（数据未显示）。饲养时C57BL / 6小鼠比FVBN小鼠更积极和频繁跳上筒的轮缘之前爬出来。如果老鼠试图通过跳跃逃跑更高的钢瓶应该被使用。 </li></ol></li></ol> 7.内皮素-1手术及脑梗塞体积的测量<ol><li>根据此前公布的协议执行4内皮素-1手术和梗死体积的测量。为目标的前部前肢运动皮层，将每只小鼠应该接受3 ET-1注射在以下坐标：（ⅰ）0.7 anteriorposterior（AP）/1.5内侧 - 外侧（毫升）/ - 1.2背腹（DV），（ ⅱ）+0.4 AP /1.25毫升/ - 1.2 DV和（ⅲ）0.1 AP /1.75毫升/ -1.2 DV 4。 </li></ol> 8.统计分析<ol><li>建议方差双向重复测量分析（ANOVA）来分析爪拖动跨不同的时间点的影响和未受影响的爪子的百分比。 </li></ol>

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量化爪拖动行为时，在气缸测试有以下要点建立：我）量化数PAW-拖动与总爪子触摸脑损伤之前，每个爪子建立一个基线;二）量化爪拖动与总爪子触摸每个爪子下面的缺血性损伤的数量;和iii）一个爪拖动并沿气缸壁的爪子的期间小鼠的躯干的横向转动的横向运动区分。 PAW-拖动是出现损伤后的前肢感觉运动皮层的新型行为。的爪式拖行为的外观因此可以用作该前肢感觉皮层已...

神经再生战略的目标是展示两个组织修复和功能的恢复。功能恢复通常评估与测量功能缺陷，在这种情况下，涉及了与损害特定脑区域相关联的运动技能行为测试。创伤性脑损伤或脑皮层的感觉运动前肢区局部缺血损伤可以由许多行为测试来证明。一个这样的试验中，在气缸试验被广泛使用的大鼠，以评估在前肢活动1的功能缺陷。测试具有低的建立费用，仅需要一个气缸，照相机和表用透明顶部。这是很容易，因为它是基于啮齿动物的自然探索行为来管理，所以训练前和食物剥夺或奖励不是必需的。尽管有这些众多的优势，在气缸测试下，利用下面的焦点受伤的前肢s到评估小鼠前肢赤字ensorimotor皮层，这是我们归因于对鼠标行为的气瓶检验分析。前肢不对称是经典测度分析气缸测试。当放置在气缸中，啮齿动物，自然由饲养到其后肢和接触汽缸壁与他们的前肢的爪子对姿势平衡探索汽缸的壁。爪接触与壁与每个前肢的数目很容易被这种探索气缸期间拍摄啮齿类定量。当受影响的前肢的爪子，使更少的接触与墙体比未受影响前肢的爪子，表明损坏对侧感觉皮层发生前肢不对称。在大鼠中，血管收缩剂的内部皮质注射剂，内皮素（ET-1），进前肢感觉皮层引起局灶性缺血损伤，这导致行为缺陷在对侧前肢。在对侧前肢使用赤字很容易检测到的变化f中在大鼠1-3缸测试orelimb不对称。在对比然而大鼠，改变前肢不对称是可变的，在小鼠中以下可比的ET-1注射4-6不太一致。在这里，我们展示了一个新的汽缸测试前肢行为分析 - 中爪拖行为分析。我们以前曾表明，爪拖动分析是损坏的小鼠比经典前肢不对称分析前肢感觉皮层的更灵敏的测量，因此是适用于各种局灶性皮质损伤模型。 检查如何前爪接触汽缸壁以下的前肢感觉皮层缺血损伤揭示了一种新的行为在小鼠-爪拖动4。当鼠标站在其后腿探索气缸壁，然后沿筒​​壁对其中线或向下的墙面上，同时拖动其受影响（禁忌病变）发生足一爪子拖它的前爪不受影响提供靠墙的姿势支持。 PAW-拖动很少发生在未受伤的老鼠因此爪子拖动的外观是受伤的前肢感觉运动皮层4正面指标。我们先前量化以下的前肢感觉皮层ET-1的局部缺血损伤的小鼠爪拖动行为和表现出持续爪拖动行为小鼠长达两周中风后4。在这里，我们表明，爪拖动行为是持续长达四周后中风。爪的拖行为分析提供了一种新的和敏感的工具，以评估在小鼠前肢感觉运动皮层局灶性缺血性脑损伤。以其物美价廉的设置，便于管理和得分使这个简单，但非常有用的工具，以快速评估在小鼠前肢行为缺陷。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Plexi-glass cylinder</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>17.5cm high, 9.5cm outer diameter, 8.8cm inner diameter, wall thickness 0.35cm (or 3.5 mm)</td>
		</tr>
		<tr>
			<td>viewing table</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>54x56x66.5cm (width x length x height), top of table is a 51x51cm sheet of plexiglass.</td>
		</tr>
		<tr>
			<td>mirror</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>34x58cm mirror</td>
		</tr>
		<tr>
			<td>video camera</td>
			<td>Sony</td>
			<td>DCR-SR42</td>
			<td>Video camera with onboard storage, SD functionality, 40x optical zoom</td>
		</tr>
		<tr>
			<td>computer</td>
			<td>Dell</td>
			<td>Optiplex 760</td>
			<td>Processor: Intel, 3.0 GHz, Memory 4.00GB (RAM)&#160;</td>
		</tr>
		<tr>
			<td>computer monitor</td>
			<td>Samsung</td>
			<td>S22C350H</td>
			<td></td>
		</tr>
		<tr>
			<td>Excel (Microsoft Office Professional Plus)</td>
			<td>Microsoft</td>
			<td>v14.0.7106.5003</td>
			<td></td>
		</tr>
		<tr>
			<td>VLC Media Player</td>
			<td>Video LAN</td>
			<td>v2.1.2</td>
			<td>Media player with playback speed modulation and video support</td>
		</tr>
		<tr>
			<td>External Hard Drive</td>
			<td>Western Digital</td>
			<td>WDBAAU0020HBK-01</td>
			<td>2 TB</td>
		</tr>
	</tbody>
</table>

paw-dragging: a novel, sensitive analysis of the mouse cylinder test

气缸试验常规地用于预测给前肢运动皮层在啮齿类动物局灶性缺血损伤。当放置在气缸中，啮齿动物探索由饲养和触摸气缸的壁与他们的前肢的爪子对姿势支承。缺血性损伤的前肢感觉运动皮层，老鼠更多地依赖于他们的前肢不受影响的爪子导致其受影响的爪子被称为前肢不对称较少触及姿势的支持。与此相反，在小鼠脑局灶性缺血损伤未能导致前肢不对称可比一致赤字。虽然前肢不对称赤字很少观察到，老鼠也表现出了一种新的行为中风后称为“爪拖”。 PAW-拖动是鼠标沿筒壁，而不是直接从墙壁从后方拆卸到一个四条腿的立场时，推下拖动其受影响的爪子的倾向。之前我们已经表明，PAW-拖动行为是小皮质缺血性损伤的前肢运动皮层高度敏感。在这里，我们为爪拖分析了详细的方案。我们定义什么是爪子拖就是和演示如何量化爪拖行为。气缸试验是一种简单且廉价的检验管理，并且不需要预训练或食物剥夺策略。在使用爪拖动分析与汽缸测试，它填补利基预测皮质缺血性损伤如photothrombosis和内皮素-1（ET-1）诱导的缺血 - 两个模型被不断增加的流行度，并产生更小的焦距受伤比大脑中动脉闭塞。最后，测量气缸测试爪拖动行为将使功能恢复使用的转基因小鼠品系，其中前面的前肢不对称分析未能检测到一致的赤字广泛队列皮质损伤后的研究。

之前我们已经证实看来，爪子拖行为遵循局灶性缺血损伤的前肢感觉运动皮层和损坏4的正面指标。 ET-1的内皮层注射入前肢感觉皮层被用来诱导局部缺血损伤（ 图8A，B）。本研究的爪子拖动行为是否延期超过14天受伤后其潜在用途，以评估功能恢复。小鼠在当日气缸试验前的ET-1注射的手术前的时间点，并在天1，3，7，14，21和28后的损伤进行测试。在每个时间点，爪子触摸和?...

Watch this Scientific Journal Video about Paw-Dragging: a Novel, Sensitive Analysis of the Mouse Cylinder Test at JoVE.com

Paw-Dragging: a Novel, Sensitive Analysis of the Mouse Cylinder Test

气缸测试的经典前肢不对称分析常规地用于评估脑损伤或中风大鼠行为缺陷;但是，它无法检测一致赤字小鼠。这项研究表明，量化爪拖动行为是脑损伤的小鼠更敏感的分析。

PAW-拖动：一本小说，鼠标缸试验敏感性分析

The cylinder test is routinely used to predict focal ischemic damage to the forelimb motor cortex in rodents. When placed in the cylinder, rodents explore ...

paw-dragging-a-novel-sensitive-analysis-of-the-mouse-cylinder-test

Research

JoVE Journal

Behavior

23.1K 次观看.  Memorial University of Newfoundland. 气缸测试的经典前肢不对称分析常规地用于评估脑损伤或中风大鼠行为缺陷;但是，它无法检测一致赤字小鼠。这项研究表明，量化爪拖动行为是脑损伤的小鼠更敏感的分析。 

视频: Paw-Dragging: a Novel, Sensitive Analysis of the Mouse Cylinder Test

Inchworming: A Novel Motor Stereotypy in the BTBR T+ Itpr3tf/J Mouse Model of Autism

Autism Spectrum Disorder (ASD) is a behaviorally defined neurodevelopmental disorder characterized by decreased reciprocal social interaction, abnormal communication, and repetitive behaviors with restricted interest. As diagnosis is based on clinical criteria, any potentially relevant rodent models of this heterogeneous disorder should ideally recapitulate these diverse behavioral traits. The BTBR T+ Itpr3tf/J (BTBR) mouse is an established animal model of ASD, displaying repetitive behaviors such as increased grooming, as well as cognitive inflexibility. With respect to social interaction and interest, the juvenile play test has been employed in multiple rodent models of ASD. Here, we show that when BTBR mice are tested in a juvenile social interaction enclosure containing sawdust bedding, they display a repetitive synchronous digging motion. This repetitive motor behavior, referred to as &#34;inchworming,&#34; was named because of the stereotypic nature of the movements exhibited by the mice while moving horizontally across the floor. Inchworming mice must use their fore- and hind-limbs in synchrony to displace the bedding, performing a minimum of one inward and one outward motion. Although both BTBR and C56BL/6J (B6) mice exhibit this behavior, BTBR mice demonstrate a significantly higher duration and frequency of inchworming and a decreased latency to initiate inchworming when placed in a bedded enclosure. We conclude that this newly described behavior provides a measure of a repetitive motor stereotypy that can be easily measured in animal models of ASD.

Inchworming: A Novel Motor Stereotypy in the BTBR T+ Itpr3tf/J Mouse Model of Autism

Inchworming is a highly repetitive synchronous digging motion displayed by BTBR T+ Itpr3tf/J (BTBR) mice when placed in a testing cage with sufficient sawdust bedding. The procedure is a modification of the juvenile social interaction protocol and is used here to assess repetitive motor stereotypies relevant to Autism Spectrum Disorder.

Inchworming：一种新型电机刻板的BTBR<em&gt; T<sup&gt; +</sup&gt; Itpr3<sup&gt; TF</sup</em自闭症&gt; /系小鼠模型

Autism Spectrum Disorder (ASD) is a behaviorally defined neurodevelopmental disorder characterized by decreased reciprocal social interaction, abnormal ...

科研

行为学

Contextual and Cued Fear Conditioning Test Using a Video Analyzing System in Mice

The contextual and cued fear conditioning test is one of the behavioral tests that assesses the ability of mice to learn and remember an association between environmental cues and aversive experiences. In this test, mice are placed into a conditioning chamber and are given parings of a conditioned stimulus (an auditory cue) and an aversive unconditioned stimulus (an electric footshock). After a delay time, the mice are exposed to the same conditioning chamber and a differently shaped chamber with presentation of the auditory cue. Freezing behavior during the test is measured as an index of fear memory. To analyze the behavior automatically, we have developed a video analyzing system using the ImageFZ application software program, which is available as a free download at http://www.mouse-phenotype.org/. Here, to show the details of our protocol, we demonstrate our procedure for the contextual and cued fear conditioning test in C57BL/6J mice using the ImageFZ system. In addition, we validated our protocol and the video analyzing system performance by comparing freezing time measured by the ImageFZ system or a photobeam-based computer measurement system with that scored by a human observer. As shown in our representative results, the data obtained by ImageFZ were similar to those analyzed by a human observer, indicating that the behavioral analysis using the ImageFZ system is highly reliable. The present movie article provides detailed information regarding the test procedures and will promote understanding of the experimental situation.

This article presents a protocol for a contextual and cued fear conditioning test using a video analyzing system to assess fear learning and memory in mice.

背景和线索恐惧制约试验小鼠中使用视频分析系统

The contextual and cued fear conditioning test is one of the behavioral tests that assesses the ability of mice to learn and remember an association ...

The Forced Swim Test as a Model of Depressive-like Behavior

The goal of the present protocol is to describe the forced swim test (FST), which is one of the most commonly used assays for the study of depressive-like behavior in rodents. The FST is based on the assumption that when placing an animal in a container filled with water, it will first make efforts to escape but eventually will exhibit immobility that may be considered to reflect a measure of behavioral despair. This test has been extensively used because it involves the exposure of the animals to stress, which was shown to have a role in the tendency for major depression. Additionally, the FST has been shown to share some of the factors that are influenced or altered by depression in humans, including changes in food consumption, sleep abnormalities and drug-withdrawal-induced anhedonia. The main advantages of this procedure are that it is relatively easy to perform and that its results are easily and quickly analyzed. Moreover, its sensitivity to a broad range of antidepressant drugs that makes it a suitable screening test is one of the most important features leading to its high predictive validity. Despite its appeal, this model has a number of disadvantages. First, the issue of chronic augmentation is problematic in this test because in real life patients need to be treated for at least several weeks before they experience any relief from their symptoms. Last, due to the aversiveness of the FST, it is important to take into account possible influences it might have on brain structure/function if brain analyses are to be carried out following this procedure.

This protocol describes the forced swim test, which is used for the study of depressive-like behavior in rodents. This procedure involves placing an animal in a container filled with water that eventually will lead to the exhibition of immobility behavior, which is considered to reflect behavioral despair.

在强迫游泳试验作为抑郁样行为的模型

The goal of the present protocol is to describe the forced swim test (FST), which is one of the most commonly used assays for the study of depressive-like ...

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.

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, ...

A Novel Approach to Assess Motor Outcome of Deep Brain Stimulation Effects in the Hemiparkinsonian Rat: Staircase and Cylinder Test

Deep brain stimulation of the subthalamic nucleus is an effective treatment option for Parkinson&#39;s disease. In our lab we established a protocol to screen different neurostimulation patterns in hemiparkinsonian (unilateral lesioned) rats. It consists of creating a unilateral Parkinson&#39;s lesion by injecting 6-hydroxydopamine (6-OHDA) into the right medial forebrain bundle, implanting chronic stimulation electrodes into the subthalamic nucleus and evaluating motor outcomes at the end of 24 hr periods of cable-bound external neurostimulation. The stimulation was conducted with constant current stimulation. The amplitude was set 20% below the individual threshold for side effects. The motor outcome evaluation was done by the assessment of spontaneous paw use in the cylinder test according to Shallert and by the assessment of skilled reaching in the staircase test according to Montoya. This protocol describes in detail the training in the staircase box, the cylinder test, as well as the use of both in hemiparkinsonian rats. The use of both tests is necessary, because the staircase test seems to be more sensitive for fine motor skill impairment and exhibits greater sensitivity to change during neurostimulation. The combination of the unilateral Parkinson model and the two behavioral tests allows the assessment of different stimulation parameters in a standardized way.

Deep brain stimulation (DBS) is an effective treatment option for Parkinson's disease. We established a study design to screen novel stimulation paradigms in rats. The protocol describes the use of the staircase test and cylinder test for motor outcome assessment in DBS treated hemiparkinsonian rats.

一种新的方法，以评估在帕金森病大鼠脑深部电刺激效应的运动结果：楼梯和缸测试

Deep brain stimulation of the subthalamic nucleus is an effective treatment option for Parkinson&#39;s disease. In our lab we established a protocol to ...

The Treadmill Fatigue Test: A Simple, High-throughput Assay of Fatigue-like Behavior for the Mouse

Fatigue is a prominent symptom in many diseases and disorders and reduces quality of life for many people. The lack of clear pathogenesis and failure of current interventions to adequately treat fatigue in all patients leaves a need for new treatment options. Despite the therapeutic need and importance of preclinical research in helping identify promising novel treatments, few preclinical assays of fatigue are available. Moreover, the most common preclinical assay used to assess fatigue-like behavior, voluntary wheel running, is not suitable for use with some strains of mice, may not be sensitive to drugs that reduce fatigue, and has relatively low throughput. The current protocol describes a novel, non-voluntary preclinical assay of fatigue-like behavior, the treadmill fatigue test, and provides evidence of its efficacy in detecting fatigue-like behavior in mice treated with a chemotherapy drug known to cause fatigue in humans and fatigue-like behavior in animals. This assay may be a beneficial alternative to wheel running, as fatigue-like behavior and potential interventions can be assessed in a greater number of mice over a shorter time frame, thus permitting faster discovery of new therapeutic options.

Fatigue is a common, undertreated and frequently poorly-understood symptom in many diseases and disorders. New preclinical assays of fatigue may help to improve current understanding and future treatment of fatigue. To that end, the current protocol provides a novel means of measuring fatigue-like behavior in the mouse.

跑步机疲劳试验：疲劳类似的行为，为鼠标操作简单，高通量检测

Fatigue is a prominent symptom in many diseases and disorders and reduces quality of life for many people. The lack of clear pathogenesis and failure of ...

The 4 Mountains Test: A Short Test of Spatial Memory with High Sensitivity for the Diagnosis of Pre-dementia Alzheimer's Disease

This protocol describes the administration of the 4 Mountains Test (4MT), a short test of spatial memory, in which memory for the topographical layout of four mountains within a computer-generated landscape is tested using a delayed match-to-sample paradigm. Allocentric spatial memory is assessed by altering the viewpoint, colors and textures between the initially presented and target images.
Allocentric spatial memory is a key function of the hippocampus, one of the earliest brain regions to be affected in Alzheimer&#39;s disease (AD) and impairment of hippocampal function predates the onset of dementia. It was hypothesized that performance on the 4MT would aid the diagnosis of predementia AD, which manifests clinically as Mild Cognitive Impairment (MCI).
The 4MT was applied to patients with MCI, stratified further based on cerebrospinal fluid (CSF) AD biomarker status (10 MCI biomarker positive, 9 MCI biomarker negative), and with mild AD dementia, as well as healthy controls. Comparator tests included tests of episodic memory and attention widely accepted as sensitive measures of early AD. Behavioral data were correlated with quantitative MRI measures of the hippocampus, precuneus and posterior cingulate gyrus.
4MT scores were significantly different between the two MCI groups (p = 0.001), with a test score of &#8804;8/15 associated with 100% sensitivity and 78% specificity for the classification of MCI with positive AD biomarkers, i.e., predementia AD. 4MT test scores correlated with hippocampal volume (r = 0.42) and cortical thickness of the precuneus (r = 0.55).
In conclusion, the 4MT is effective in identifying the early stages of AD. The short duration, easy application and scoring, and favorable psychometric properties of the 4MT fulfil the need for a simple but accurate diagnostic test for predementia AD.

The 4 Mountains Test: A Short Test of Spatial Memory with High Sensitivity for the Diagnosis of Pre-dementia Alzheimer&#039;s Disease

This article describes the 4 Mountains Test (4MT), a hippocampus-dependent test of working allocentric spatial memory. The hippocampus is affected early in Alzheimer's disease (AD) and this article outlines the 4MT methodology and results of patient testing, which demonstrates the value of the 4MT in the diagnosis of pre-dementia AD.

四山测试：空间记忆的一个简短的测试与高灵敏度的预痴呆诊断阿尔茨海默氏病

This protocol describes the administration of the 4 Mountains Test (4MT), a short test of spatial memory, in which memory for the topographical layout of ...

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

An individual's sexual drive is influenced by genotype, experience and environmental conditions. How these factors interact to modulate sexual behaviors remains poorly understood. In Drosophila melanogaster, environmental cues, such as food availability, affect mating activity offering a tractable system to investigate the mechanisms modulating sexual behavior. In D. melanogaster, environmental cues are often sensed via the chemosensory gustatory and olfactory systems. Here, we present a method to test the effect of environmental chemical cues on mating behavior. The assay consists of a small mating arena containing food medium and a mating couple. The mating frequency for each couple is continuously monitored for 24 h. Here we present the applicability of this assay to test environmental compounds from an external source through a pressurized air system as well as manipulation of the environmental components directly in the mating arena. The use of a pressurized air system is especially useful to test the effect of very volatile compounds, while manipulating components directly in the mating arena can be of value to ascertain a compound's presence. This assay can be adapted to answer questions about the influence of genetic and environmental cues on mating behavior and fecundity as well as other male and female reproductive behaviors.

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

We demonstrate an assay to analyze the environmental and genetic cues that influence mating behavior in the fruit fly Drosophila melanogaster.

测试环境线索对交配行为的影响的方法<em&gt;黑腹果蝇</em

An individual's sexual drive is influenced by genotype, experience and environmental conditions. How these factors interact to modulate sexual behaviors ...

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

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 fairly simple and can be completed over 3 days: habituation day, training day, and testing day. During training, the mouse is allowed to explore 2 identical objects. On test day, one of the training objects is replaced with a novel object. Because mice have an innate preference for novelty, if the mouse recognizes the familiar object, it will spend most of its time at the novel object. Due to this innate preference, there is no need for positive or negative reinforcement or long training schedules. Additionally, the ORT can also be modified for numerous applications. The retention interval can be shortened to examine short-term memory, or lengthened to probe long-term memory. Pharmacological intervention can be used at various times prior to training, after training, or prior to recall to investigate different phases of learning (i.e., acquisition, early or late consolidation, or recall). Overall, the ORT is a relatively low-stress, efficient test for memory in mice, and is appropriate for the detection of neuropsychological changes following pharmacological, biological, or genetic manipulations.

The object recognition test (ORT) is a simple and efficient assay for evaluating learning and memory in mice. The methodology is described below.

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 ...

An Instrumented Pull Test to Characterize Postural Responses

Impairment of postural reflexes, termed postural instability, is a common and disabling deficit in Parkinson's disease. To assess postural reflexes, clinicians typically employ the pull test to grade corrective responses to a backward perturbation at the shoulders. However, the pull test is prone to issues with reliability and scaling (score/4). Here, we present an instrumented version of the pull test to more precisely quantify postural responses. Akin to the clinical test, pulls are manually administered except pull force is also recorded. Displacements of the trunk and feet are captured by a semi-portable motion tracking system. Raw data represent distance traveled (in millimeter units), making subsequent interpretation and analysis intuitive. The instrumented pull test also detects variabilities influencing pull test administration, such as pull force, thereby identifying and quantifying potential confounds that can be accounted for by statistical techniques. The instrumented pull test could have application in studies seeking to capture early abnormalities in postural responses, track postural instability over time, and detect responses to therapy.

Impairment of postural reflexes, termed postural instability, is difficult to quantify. Clinical assessments such as the pull test suffer issues with reliability and scaling. Here, we present an instrumented version of the pull test to objectively characterize postural responses.

一种表征姿势反应的仪器拉力测试

Impairment of postural reflexes, termed postural instability, is a common and disabling deficit in Parkinson's disease. To assess postural reflexes, ...