多亏了 Rassam 的样本收集, 伊迪丝 Klobetz--对 FCM 的分析, PD 博士. 兽医。habil。萝丝维塔梅尔协助统计分析, 并 Wiebke Gentner 校对手稿。这项研究是柏林-勃兰登堡研究平台 BB3R (www.bb3r.de) 的一部分, 由德国联邦教育部和研究部资助 (赠款号: 031A262A) (www.bmbf.de/en/index.html)。

这项研究是根据《德国动物福利法》规定的准则进行的, 并得到了柏林国家当局 ("Landesamt 并且一厢情愿和 Soziales") 的批准, 允许号码: G0053/15)。
注: 本议定书的主要目的是研究反复麻醉对糖皮质激素代谢物的影响。为了确定要使用的动物数量, 进行了一个样本大小计算: n ≥ 2 x (s/µ1-µ2)2 x (zα + zβ)2。µ1-µ2是指在其中进行功率和样本大小计算 (α = 5%, β = 80%) 的填充方式之间的差异; zα = 1.96 和 zβ = 0.84 是标准正态分布的分位数。图 1说明了此协议的时间线。如果协议的参数显示与控制级别的差异, 应密切监视动物, 并在适当的时间后再测量参数。例如, 如果增加了特质焦虑相关行为, 则应在一周后再次测试此行为, 以帮助确定期间直到完全恢复。本协议中定义的时间点和周期可用于其他过程。在更改时间点时, 应按照协议中的说明来保留适应期。为了减少可能影响小鼠行为的因素, 需要进行更多操作的测试应在不干扰小鼠正常行为的试验后进行。图 2使用摘要计分单汇总了协议的所有测试。图 3提供了福利等级的简化刻度, 它概述了如何解释测试结果。
1. Habituating 老鼠处理的实验者
<ol><li>允许老鼠习惯到动物设施至少2周后, 他们已经从其他设施或供应商获得。</li>
	<li>小鼠在小组和维护他们在标准条件下 (室温 22 2 °c; 相对湿度 55 * 10%) 在光: 黑暗的周期 12:12 h。</li>
	<li>为所有组提供隧道和棉花 nestlets 作为标准浓缩, 并提供食物和水的ad 随意。</li>
	<li>在测试12之前, 将所有鼠标习惯到隧道和/或杯处理至少一周。 注意: 通过尾部拾取鼠标可以诱发压力或焦虑, 进而影响福祉, 也会对此协议12的结果产生影响。</li>
</ol>2. 准备行为测试室和仪器
注意: 提供一个单独的测试空间, 最好是在保存动物的房间附近。在手术前至少60分钟将老鼠送到试验室。如果可能, 在执行该过程的同一测试室中进行本协议的所有测试。
<ol><li>准备一个观察笼来测试穴居行为8 , 并拍摄照片, 以用于在 mg13 (图 4)。<ol>
<li>使用一个玻璃盒, 建筑面积约220毫米 x 290 毫米, 高度为390毫米。</li>
		<li>用大约0.5 厘米的寝具材料盖住这个盒子的地板。</li>
		<li>在新的床上用品材料的顶部, 将一小部分使用过的床上用品从家庭笼子里撒出来, 以减少新环境造成的痛苦。</li>
		<li>提供食物, 通常作为饮食和水供应的同类食品。 注: 如有可能, 使用水瓶, 因为老鼠可以用床上用品填充水碗。</li>
	</ol>
</li>
	<li>为24小时观察期间准备一个保持架 (III 型: 420 毫米 x 260 毫米 x 150 毫米), 为其单独存放鼠标 (图 5)。 注: 为了最大限度地减少个体住房的持续时间, 在这一时期收集巢楼行为、家庭笼活动、食物摄取和粪便皮质酮代谢物 (FCM) 的数据。<ol>
<li>将新的床上用品材料放在笼中 (大约0.5 厘米深), 并在新材料的顶部分散一小部分使用过的寝具材料, 而不粪便从主笼上, 以减少遇险。</li>
		<li>提供标准的方形棉 nestlet 的定义的重量, 作为环境浓缩仅 (参见材料表)14。 注意: 商业 nestlets 的重量可能不同。因此, 我们修改了执事描述的 nestlet 的重量, 使用了 2.0 g 而不是 2.7 g14。</li>
		<li>在保持架顶部安装红外线传感器时, 使用红外线传感器测量主笼活动 (请参阅材料表)。</li>
		<li>提供食物, 通常作为饮食供应的同类, 和水ad 随意。</li>
	</ol>
</li>
</ol>3. 鼠标鬼脸秤
注: 在观察笼中拍摄的三个时间点的照片: (i) 在程序前2天记录基线的标准, (ii) 30 分钟后的程序, 和 (iii) 150 分钟后的程序。当福利被削弱时, 他的分数就会增加。如果在150分钟后仍能观察到增加的。
<ol><li>使用高清相机拍摄照片。</li>
	<li>将鼠标轻轻地转移到观察笼中, 让鼠标习惯到新的环境中至少30分钟。</li>
	<li>在 1-2 分钟内连续拍摄每段时间的 30-40 张照片。</li>
	<li>对所有照片进行排序, 选择尖锐的正面或侧面照片, 丢弃模糊的照片或照片, 显示鼠标面对从其他角度比正面或侧面的看法。</li>
	<li>从每个时间点随机选择一张照片, (即过程前2天, 程序后30分钟, 以及过程后150分钟) 为每个鼠标。</li>
	<li>裁剪照片以仅显示鼠标的头部, 以使正文位置不可见13。</li>
	<li>为每张照片创建一个电子表格文件, 并在每张纸上添加一个表, 包括五面部动作单元。 注: 该文件包含基线照片和照片后的过程。</li>
	<li>随机化工作表的顺序。</li>
	<li>将计算机屏幕上的文件显示给三名独立人员, 这些人以前受过训练, 使用由兰福德et . 开发的 "特里", 并让他们用3点的比例 (0 = 不存在, 1 = 适度存在, 2 = 明显) 得分面部动作单位。目前)。 注: 评分依据的是以下参数13: 轨道收紧 ("眶面积缩小, 眼睑紧密闭合或眼挤压");鼻子隆起 ("在鼻子的桥梁可看见的被圆的延伸皮肤);面颊凸起 ("面颊肌肉的凸的出现);耳朵位置 ("耳朵从基线位置分离和向后, 或具有垂直脊, 形成由于耳朵被拉回的提示");晶须变化 ("晶须从基线位置向后、正面或向前移动, 仿佛站在末端;胡须也可能聚集在一起 ")。</li>
	<li>分析分数, 如下所示 (从兰福德et 。13)。<ol>
<li>平均所有的面部动作单位为每张照片, 以产生的 "中" 得分。 注: 如果一个面部动作单位不能得分, 平均其余的面部动作单位。</li>
		<li>减去平均值的基线照片从平均值的照片后的过程, 以获得一个不同的分数为每一个鼠标。</li>
		<li>测试不同的人之间的差异评分 (非参数测试的相关样本)。 注: 如果有显著差异 (p < 0.05), 确定是否所有照片的分数或只有几张照片的分数不同的人之间。如果后者是真的, 重复评分这些照片。否则, 这些人应该重复训练, 然后再拍照片。</li>
		<li>如果所有的人的结果没有显著的差异, 平均每只老鼠的不同得分者的差异评分。</li>
		<li>使用非参数统计检验, 比较研究组之间平均差异评分。</li>
	</ol>
</li>
</ol>4. 穴居行为8,15,16
<ol><li>在标准不透明的塑料水瓶 (250 毫升, 150 毫米长度, 55 毫米直径, 45 毫米直径的瓶子颈) 上放置 140 "2 克" 的食物颗粒, 以此来准备洞穴8。 注意: 小鼠喜欢宽管, 有68毫米直径的洞穴可以按照执事16的描述使用。</li>
	<li>在驯化的过程前5天, 将洞穴填满食物颗粒在笼子里。 注意: 笼子里的常规食品分配单位不应该被清空, 但也应该留满食物颗粒, 因为老鼠已经习惯了。</li>
	<li>进行两次测试, 2 天前的程序 (基线);执行最后30分钟的开机自检程序。<ol>
<li>让鼠标习惯至少30分钟的观察笼, 在那里拍摄的照片为的。</li>
		<li>将塑料水壶装满与观察笼的后壁平行的食物小球。</li>
		<li>在2小时后的洞穴中, 称食物颗粒 (g)。</li>
	</ol>
</li>
	<li>计算相对于初始重量的小鼠从洞穴中取出的食物颗粒的重量 (%)。</li>
</ol>5. 24 小时观察期
注意: 老鼠单独安置, 如2.2 所述。(图 5), 为期24小时, 以测量食物摄取量、家庭笼活动、巢建筑行为和 FCM 水平。24 h 的观察发生两次: (一) 在程序开始之日前2天 (二)。
<ol><li>
食物摄取量
	<ol>
<li>定期对老鼠进行称量 (例如麻醉前2天, 麻醉前、麻醉后2天、麻醉后的每周), 以评估体重的变化 (评分表的一部分)。 注意: 体重需要计算每克体重的食物摄取量。在24小时观察期间也可以测量进水量。如果食物摄入量减少, 福利可能会受损。</li>
		<li>确定笼中食物单位 (约100克) 所提供的标准食物饮食 (克) 的初始重量。</li>
		<li>在24小时的观察期结束时确定标准饮食的重量。</li>
		<li>仔细扫描食物单位下方的笼侧, 以便食物溢出, 并添加任何额外的食物颗粒, 发现食品颗粒剩余的食物单位的重量。</li>
		<li>计算每单位体重的食物摄取量。</li>
	</ol>
</li>
	<li>
家庭笼活动 注: 以下说明是指使用红外线传感器 (请参阅材料表), 但家庭笼子活动也可以用其他程序评估。家庭笼活动从控制级别的偏差 (例如hypoactivity、多动症) 可能是幸福度受损的标志。<ol>
<li>启动程序。</li>
		<li>选择一个采样间隔为1分钟和一个采集时间为24小时, 这意味着脉冲记录每分钟24小时。 注: 如果实验者在录制开始后几次进入房间, 则只使用句点中的数据, 当鼠标未受到干扰时 (即在黑暗期间)。</li>
		<li>总结10分钟的脉冲间隔。</li>
		<li>计算时间曲线下的区域 (脉冲 x min)。</li>
	</ol>
</li>
	<li>
巢大厦行为 注意: 复杂和高巢可以作为福祉的指标。<ol>
<li>放置一个方形棉花 nestlet (参见材料表), 其定义的重量 (例如2.0 g) 在笼子中间。</li>
		<li>根据执事14在第二天早晨, 大约2小时在光打开以后, 在5点刻度上评分巢 (如下所示)。权衡任何 untorn nestlet 件, 至少是最初 nestlet 重量的5%。将巢分如下14<ol>
<li>如果90% 的 nestlet 完好无损, 则分配 "1" 分。</li>
			<li>指定分数为 "2", 如果它是 50-90% 完好无损。</li>
			<li>分配分数 "3", 如果 50-90% 的 nestlet 被粉碎。</li>
			<li>分配评分 "4", 如果超过90% 被粉碎, 但巢是平的, 少于50% 的周长比鼠标身高时, 蜷缩起来。</li>
			<li>分配评分 "5", 如果超过 90% nestlet 被撕碎和巢是高, 超过50% 的周长高于身高的一个卷曲的鼠标。</li>
		</ol>
</li>
	</ol>
</li>
	<li>
粪皮质酮代谢物 注意: 在控制水平以上的 FCM 的增加反映了24小时 postanesthetic 期间的急性应激水平。<ol>
<li>在24小时观察期结束时使用镊子从笼中收集所有干粪丸, 并清除尿液中的湿颗粒。</li>
		<li>提取 FCM 根据棕榈, et 等。17, 如下所示。<ol>
<li>干燥粪样品在温度 60-70 °c。</li>
			<li>用灰浆融汇粪便样品。</li>
			<li>将0.05 克的整除与1毫升80% 甲醇在离心管中摇动, 在多涡上进行30分钟。</li>
			<li>离心机样品在 2500 x g 15 分钟。</li>
			<li>将0.5 毫升的上清液注入另一离心管。</li>
			<li>储存粪便样品 (和提取物) 在至少-18 摄氏度。</li>
			<li>用 5α-pregnane-3 b、11 b、21-三元醇-20-一酶免疫分析法 (EIA) 18、19或另一次完全验证的环境影响评估 FCM.</li>
		</ol>
</li>
		<li>计算 fcm 浓度相对于基线 fcm 浓度的百分比变化。</li>
	</ol>
</li>
</ol>6. 自由探索范式
<ol><li>把笼子从架子上拿出来, 在24小时观察期结束时把它放在桌面上。</li>
	<li>将网格化笼顶 (不带食物或水瓶) 放在位于45°角的笼中, 置于笼的较长一侧。 注意: 不要破坏巢, 它充当老鼠的藏身之处, 但将笼顶对角置于巢之上。</li>
	<li>显示器或视频-记录小鼠10分钟从距离约1.5 米。<ol>
<li>启动计时器。</li>
		<li>注意当鼠标爬到笼顶 (所有四只爪子放在笼顶) 或离开笼顶 (用一个或更多的爪子在笼底) 的所有时间。 注意: 一些老鼠可能爬上笼顶, 离开它, 以便沿着笼子的边缘走。一些老鼠也在笼子顶上。把这些病例当作老鼠仍然放在笼子顶上。</li>
	</ol>
</li>
	<li>根据伯特et计算参数。20。<ol>
<li>分析第一次探测的滞后时间 (秒)。</li>
		<li>分析探索的次数。</li>
		<li>分析勘探的总持续时间 (秒)。 注意: 第一次勘探的潜伏期很高, 探索的次数少, 总的勘探时间也很低, 这表明有较高的特质焦虑水平。</li>
	</ol>
</li>
</ol>

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作者没有什么可透露的。

该议定书最初是为了评估接受单一麻醉或重复异氟醚麻醉的 C57BL/6JRj 小鼠的福祉而制定的。研究结果证实, 对奢侈品行为的测试以及其他措施 (例如自由探索范式、mg、穴居食物摄取) 都是评估幸福感的敏感方法。反复异氟醚麻醉对特质焦虑相关行为、特征和穴居行为产生短期影响。此外, 反复异氟醚麻醉似乎影响食物摄取量10。
自由探索范式表明, 减少...

欧盟指令 2010/631规定, 每当需要动物试验时, 由罗素和2开发的3R 原理 (替换、还原、细化) 将被应用.欧盟指令的最终目标是逐步淘汰所有动物试验, 但该指令承认, 目前还需要一些动物实验来进行研究, 以保护人类和动物的健康。因此, 如果动物实验不能被任何替代方法所取代, 只有最小数量的实验动物被用来获得可靠的结果。此外, 使用适当的细化措施, 应尽量减少伴随实验程序的疼痛、痛苦和苦恼的数量。欧盟指令2010/63 规定, 过程的严重性必须前瞻性归类为非恢复、温和、适度或严重的1。由于严重性分类是根据个案来决定的, 因此有科学的声音工具来估计给定过程的严重性是很重要的。
根据莫顿和格里菲斯的建议, 评分表3是检测任何偏离正常状态的重要工具, 包括对福祉4的负面影响。评分表用于回顾性地确定由实验引起的疼痛、痛苦和苦恼, 并着重于个体动物 (例如、体重、皮毛、步态) 的物理状态的可见变化。虽然, 欧盟指令2010/63 的附件八提供了每个严重性类别的例子, 但研究人员仍然缺乏利用科学基础的数据来估计给定程序严重性程度的工具。
缺乏显示消极福祉的指标并不是确定动物状况的唯一方法;指向正福利的指示器的存在也很重要5、6、7、8。例如, 动物只在满足所有基本需求时, 才会表现出诸如挖洞和筑巢等奢侈行为。如果减少了福利, 奢侈品行为首先会下降5,7。用于评估福祉的议定书应包括指示动物生理、生理/生物化学和心理状态的指标, 以便以详细和全面的方式对其福祉进行评估9。
在改进的范围内, 制定了一项议定书, 以满足这些要求, 并评估一项涉及全麻对小鼠福祉的程序的影响10。同时, 目标是尽量减少任何额外的压力, 使协议易于集成到给定的实验中。该协议考虑穴居行为, 家庭笼行为, 如活动, 食物摄取, 和巢, 和特质焦虑相关的行为。此外, 还包括小鼠的鬼脸量表和粪便中皮质酮代谢物的无创性分析。该议定书的目的是促进以科学和动物为中心的方式评估福祉, 并提供支持严重程度分类的福祉信息。除了评分表之外, 它还可以为过程的严重性分类提供有用的信息。由于该协议易于执行且不需要广泛的设备, 因此可以将其集成到一个正在进行的实验中, 而不影响研究结果。应该指出的是, 动物研究: 报告的活体实验 (到达) 指南11将在所有涉及动物实验的研究中观察到, 目的是改进设计、分析和报告。

<table border="0" cellpadding="0" cellspacing="0" width="603">
	<tbody>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Isofluran</td>
			<td style="width:101px;">CP-Pharma Handelsgesellschaft mbH</td>
			<td style="width:119px;">1214</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">InfraMot - Sensore Units</td>
			<td style="width:101px;">TSE Systems</td>
			<td>302015-SENS</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">InfraMot - Control Units</td>
			<td style="width:101px;">TSE Systems</td>
			<td style="width:119px;">302015-C/16</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">InfraMot - Software</td>
			<td style="width:101px;">TSE Systems</td>
			<td style="width:119px;">302015-S</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Nestlet N</td>
			<td style="width:101px;">Ancare - Plexx</td>
			<td style="width:119px;">NES3600</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Camera EOS 350D</td>
			<td style="width:101px;">Canon</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

systematic assessment of well-being in mice for procedures using general anesthesia

根据罗素和科里开发的3R 原理 (替换、还原、细化), 科学研究应尽可能使用动物实验的替代品。如果不能替代动物实验, 那么所使用的实验动物总数应该是获取有价值数据所需的最低数量。此外, 应采用适当的细化措施, 以尽量减少伴随实验程序的痛苦、痛苦和痛苦。用于分类疼痛、痛苦和痛苦程度的类别是不恢复的、轻度的、中等的或严重的 (欧盟指令 2010/63)。要确定哪些类别适用于个别案例, 使用科学的声音工具是至关重要的。
这里提出的幸福评估协议是为使用全麻的程序设计的。该协议的重点是家庭笼活动, 鼠标鬼脸规模, 和奢侈行为, 如穴居和巢建筑行为作为福祉的指标。它也使用自由探索范式的特质焦虑相关行为。粪便皮质酮代谢物作为急性应激指标的测量在24小时后麻醉期间。
该议定书提供了有关全麻后小鼠的福祉的科学可靠的信息。由于其简单性, 该协议可以很容易地被修改和集成在一个计划的研究。虽然它没有提供一个规模, 根据欧盟指令2010/63 分类的苦恼, 它可以帮助研究人员估计一个程序的严重性程度, 使用科学的声音数据。它提供了一种以科学、动物为中心的方式来改进对幸福的评估的方法。

本议定书最初是为了评估 C57BL/6JRj 小鼠在单异氟醚麻醉 (一次45分钟麻醉疗程, n = 13 女性) 或重复异氟醚麻醉 (六45分钟麻醉疗程 3-4 天) 后的福祉而制定的。在麻醉疗程之间, n = 13 女性) 与控制小鼠 (n = 6 位女性) 的福利相比,10, 没有接受麻醉, 但根据相同的措施测试。我们评估了单一经验的异氟醚麻醉和重复异氟醚麻醉对 C57BL/6JRj 小鼠的幸福与未经治疗的控?...

Watch this Scientific Journal Video about Systematic Assessment of Well-Being in Mice for Procedures Using General Anesthesia at JoVE.com

Systematic Assessment of Well-Being in Mice for Procedures Using General Anesthesia

我们制定了一个协议, 以评估小鼠的福祉在程序中使用全麻。分析了一系列的行为参数, 说明了幸福水平和糖皮质激素代谢产物。该议定书可作为一般援助, 以科学的、以动物为中心的方式估计严重程度。

全身麻醉下小鼠幸福度的系统评价

In keeping with the 3R Principle (Replacement, Reduction, Refinement) developed by Russel and Burch, scientific research should use alternatives to animal ...

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JoVE Journal

Behavior

12.0K 次观看.  Freie Universität Berlin. 我们制定了一个协议, 以评估小鼠的福祉在程序中使用全麻。分析了一系列的行为参数, 说明了幸福水平和糖皮质激素代谢产物。该议定书可作为一般援助, 以科学的、以动物为中心的方式估计严重程度。

视频: Systematic Assessment of Well-Being in Mice for Procedures Using General Anesthesia

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 ...

科研

行为学

Assessment of Age-related Changes in Cognitive Functions Using EmoCogMeter, a Novel Tablet-computer Based Approach

The main goal of this study was to assess the usability of a tablet-computer-based application (EmoCogMeter) in investigating the effects of age on cognitive functions across the lifespan in a sample of 378 healthy subjects (age range 18-89 years). Consistent with previous findings we found an age-related cognitive decline across a wide range of neuropsychological domains (memory, attention, executive functions), thereby proving the usability of our tablet-based application. Regardless of prior computer experience, subjects of all age groups were able to perform the tasks without instruction or feedback from an experimenter. Increased motivation and compliance proved to be beneficial for task performance, thereby potentially increasing the validity of the results. Our promising findings underline the great clinical and practical potential of a tablet-based application for detection and monitoring of cognitive dysfunction.

We tested the usability of a tablet-computer-based application (EmoCogMeter) in investigating the effects of age on cognition. Results show an age-related cognitive decline, thereby proving the usability of our application. Findings underline the great clinical and practical potential of a tablet-based application for detection and monitoring of cognitive dysfunction.

与年龄相关的变化认知功能使用EmoCogMeter，新型平板计算机基础法评估

The main goal of this study was to assess the usability of a tablet-computer-based application (EmoCogMeter) in investigating the effects of age on ...

Operant Procedures for Assessing Behavioral Flexibility in Rats

Executive functions consist of multiple high-level cognitive processes that drive rule generation and behavioral selection. An emergent property of these processes is the ability to adjust behavior in response to changes in one&#8217;s environment (i.e., behavioral flexibility). These processes are essential to normal human behavior, and may be disrupted in diverse neuropsychiatric conditions, including schizophrenia, alcoholism, depression, stroke, and Alzheimer&#8217;s disease. Understanding of the neurobiology of executive functions has been greatly advanced by the availability of animal tasks for assessing discrete components of behavioral flexibility, particularly strategy shifting and reversal learning. While several types of tasks have been developed, most are non-automated, labor intensive, and allow testing of only one animal at a time. The recent development of automated, operant-based tasks for assessing behavioral flexibility streamlines testing, standardizes stimulus presentation and data recording, and dramatically improves throughput. Here, we describe automated strategy shifting and reversal tasks, using operant chambers controlled by custom written software programs. Using these tasks, we have shown that the medial prefrontal cortex governs strategy shifting but not reversal learning in the rat, similar to the dissociation observed in humans. Moreover, animals with a neonatal hippocampal lesion, a neurodevelopmental model of schizophrenia, are selectively impaired on the strategy shifting task but not the reversal task. The strategy shifting task also allows the identification of separate types of performance errors, each of which is attributable to distinct neural substrates. The availability of these automated tasks, and the evidence supporting the dissociable contributions of separate prefrontal areas, makes them particularly well-suited assays for the investigation of basic neurobiological processes as well as drug discovery and screening in disease models.

The ability to assess executive functions such as behavioral flexibility in rats is useful for investigating the neurobiology of cognition in both intact animals and disease models. Here we describe automated tasks for assessing strategy shifting and reversal learning, which are particularly sensitive to disruptions in prefrontal cortical networks.

操作性程序，以评估大鼠的行为灵活性

Executive functions consist of multiple high-level cognitive processes that drive rule generation and behavioral selection. An emergent property of these ...

Experimental Assessment of Mouse Sociability Using an Automated Image Processing Approach

Mouse is the preferred model organism for testing drugs designed to increase sociability. We present a method to quantify mouse sociability in which the test mouse is placed in a standardized apparatus and relevant behaviors are assessed in three different sessions (called session I, II, and III).
The apparatus has three compartments (see Figure 1), the left and right compartments contain an inverted cup which can house a mouse (called &#34;stimulus mouse&#34;).
In session I, the test mouse is placed in the cage and its mobility is characterized by the number of transitions made between compartments. In session II, a stimulus mouse is placed under one of the inverted cups and the sociability of the test mouse is quantified by the amounts of time it spends near the cup containing the enclosed stimulus mouse vs. the empty inverted cup. In session III, the inverted cups are removed and both mice interact freely. The sociability of the test mouse in session III is quantified by the number of social approaches it makes toward the stimulus mouse and by the number of times it avoids a social approach by the stimulus mouse.
The automated evaluation of the movie detects the nose of the test mouse, which allows the determination of all described sociability measures in session I and II (in session III, approaches are identified automatically but classified manually). To find the nose, the image of an empty cage is digitally subtracted from each frame of the movie and the resulting image is binarized to identify the mouse pixels. The mouse tail is automatically removed and the two most distant points of the remaining mouse are determined; these are close to nose and base of tail. By analyzing the motion of the mouse and using continuity arguments, the nose is identified.
<img alt="Figure 1" src="/files/ftp_upload/52508/52508fig1.jpg" /> 
Figure 1. Assessment of Sociability During 3 sessions. Session I (top): Acclimation of test mouse to the cage. Session II (middle): Test mouse moving freely in the cage while the stimulus mouse is enclosed in an inverted cup. Session III (bottom): Both test mouse and stimulus mouse are allowed to move freely and interact with each other.

This protocol describes a method to quantify mouse sociability. Mice are videotaped as they move and interact in a special cage. Movie processing allows for the automated quantification of sociability with excellent accuracy and reliability.

使用自动图像处理方法鼠标社交性的实验评估

Mouse is the preferred model organism for testing drugs designed to increase sociability. We present a method to quantify mouse sociability in which the ...

Assessment of Cocaine-induced Behavioral Sensitization and Conditioned Place Preference in Mice

It is thought that rewarding experiences with drugs create strong contextual associations and encourage repeated intake. In turn, repeated exposures to drugs of abuse make lasting alterations in the brain function of vulnerable individuals, and these persistent alterations likely serve to maintain the maladaptive drug seeking and taking behaviors characteristic of addiction/dependence2. In rodents, reward experience and contextual associations are frequently measured using the conditioned place preference assay, or CPP, wherein preference for a previously drug-paired context is measured. Behavioral sensitization, on the other hand, is an increase in a drug-induced behavior that develops progressively over repeated exposures. Since sensitized behaviors can often be measured after several months of drug abstinence, depending on the dose and length of initial exposure, they are considered observable correlates of lasting drug-induced plasticity. Researchers have found these assays useful in determining the neurobiological substrates mediating aspects of addiction as well as assessing the potential of different interventions in disrupting these behaviors. This manuscript describes basic, effective protocols for mouse CPP and locomotor behavioral sensitization to cocaine.

This protocol is intended to enable researchers to conduct experiments designed to test these aspects of addiction using the conditioned place preference and locomotor behavioral sensitization assays.

可卡因诱导行为敏和小鼠位置偏爱评估

It is thought that rewarding experiences with drugs create strong contextual associations and encourage repeated intake. In turn, repeated exposures to ...

Battery of Behavioral Tests Assessing General Locomotion, Muscular Strength, and Coordination in Mice

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects and outcomes that a particular disease or treatment has on the animal&#39;s wellbeing and health. There are numerous behavioral tests that may be applied. We selected a test for general locomotion, the open field test (OFT); a test for muscular strength, the mesh test (MT); and a test for coordination, the rotarod test (RR). Testing can be accomplished on a weekly or monthly basis. As a test for general locomotion, the OFT works by objectively monitoring movement parameters while the mouse is in an open field apparatus. The field is generally a 2&#39; x 2&#39; box, and the movements are recorded through laser sensing or through video capture. The mouse is placed in the center of the open field and allowed to move freely for the test. The MT uses the latency for a mouse to fall off an inverted screen as a measure of muscular strength. A mouse is placed on a screen, which is inverted over a clear box, and is timed for their latency to fall. Three trials are performed, with the best of the three trials scored for that day. A score of 60 s is the maximum time a mouse is left inverted. Mice are given a 5-min rest period between mesh test trials. Lastly, an accelerated protocol on the RR assesses motor coordination and endurance. During a trial, a mouse walks on a rotating rod as it increases in speed from 4 rpm to 40 rpm over 5 min. The trial ends when the mouse touches the magnetized pressure sensor upon falling. Each mouse undergoes three trials, and the best trial is scored for that day. This combined behavioral data allows for the global assessment of mobility, coordination, strength, and movement of the test animals. At least two out of the three behavioral testing measures must show improvement for an animal to qualify as having overall improved motor function.

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects of a disease or treatment on an animal's wellbeing. In order to globally assess motor functioning, we selected tests for general locomotion, muscular strength, and coordination: the open field test, the mesh test, and the rotarod test, respectively.

用于评估一般运动、肌肉力量和小鼠协调性的行为测试电池

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects and outcomes that a particular disease or treatment has on the ...

Tickling, a Technique for Inducing Positive Affect When Handling Rats

Handling small animals such as rats can lead to several adverse effects. These include the fear of humans, resistance to handling, increased injury risk for both the animals and the hands of their handlers, decreased animal welfare, and less valid research data. To minimize negative effects on experimental results and human-animal relationships, research animals are often habituated to being handled. However, the methods of habituation are highly variable and often of limited effectiveness. More potently, it is possible for humans to mimic aspects of the animals&#39; playful rough-and-tumble behavior during handling. When applied to laboratory rats in a systematic manner, this playful handling, referred to as tickling, consistently gives rise to positive behavioral responses. This article provides a detailed description of a standardized rat tickling technique. This method can contribute to future investigations into positive affective states in animals, make it easier to handle rats for common husbandry activities such as cage changing or medical/research procedures such as injection, and be implemented as a source of social enrichment. It is concluded that this method can be used to efficiently and practicably reduce rats&#39; fearfulness of humans and improve their welfare, as well as reliably model positive affective states.

This article demonstrates the standardized application of playful handling, a tickling technique designed to mimic rat rough-and-tumble play. This technique is effective at reducing fearful reactions to humans and generating positive affect when rats are handled for common husbandry activities and medical and research procedures such as injection.

痒, 一种诱发大鼠正向影响的技术

Handling small animals such as rats can lead to several adverse effects. These include the fear of humans, resistance to handling, increased injury risk ...

Supramaximal Intensity Hypoxic Exercise and Vascular Function Assessment in Mice

Exercise training is an important strategy for maintaining health and preventing many chronic diseases. It is the first line of treatment recommended by international guidelines for patients suffering from cardiovascular diseases, more specifically, lower extremity artery diseases, where the patients' walking capacity is considerably altered, affecting their quality of life. 
Traditionally, both low continuous exercise and interval training have been used. Recently, supramaximal training has also been shown to improve athletes' performances via vascular adaptations, amongst other mechanisms. The combination of this type of training with hypoxia could bring an additional and/or synergic effect, which could be of interest for certain pathologies. Here, we describe how to perform supramaximal intensity training sessions in hypoxia on healthy mice at 150% of their maximal speed, using a motorized treadmill and a hypoxic box. We also show how to dissect the mouse in order to retrieve organs of interest, particularly the pulmonary artery, the abdominal aorta, and the iliac artery. Finally, we show how to perform ex vivo vascular function assessment on the retrieved vessels, using isometric tension studies.

High-intensity training in hypoxia is a protocol that has been proven to induce vascular adaptations potentially beneficial in some patients and to improve athletes&#39; repeated sprint ability. Here, we test the feasibility of training mice using that protocol and identify those vascular adaptations using ex vivo vascular function assessment.

小鼠低血压运动的下位强度及血管功能评估

Exercise training is an important strategy for maintaining health and preventing many chronic diseases. It is the first line of treatment recommended by ...

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 ...

Three Laboratory Procedures for Assessing Different Manifestations of Impulsivity in Rats

The present article provides a guide for the conduction and analysis of three conditioning-based protocols to evaluate impulsivity in rats. Impulsivity is a meaningful concept because it is associated with psychiatric conditions in humans and with maladaptive behavior in non-human animals. It is believed that impulsivity is composed of separate factors. There are laboratory protocols devised to assess each of these factors using standardized automated equipment. Delay discounting is associated with the incapacity to be motivated by delayed outcomes. This factor is evaluated through intertemporal choice protocols, which consist&#160;of presenting the individual with a choice situation involving an immediate reward and a larger but delayed reward. Response inhibition deficit is associated with the incapacity to withhold prepotent responses. Differential reinforcement of low rates (DLR) and feature-negative discrimination protocols assess the response inhibition deficit factor of impulsivity. The former imposes a condition to a motivated individual in which most wait a minimum period of time for a response to be rewarded. The latter evaluates the capacity of individuals to refrain from food seeking responses when a signal of the absence of food is presented. The purpose of these protocols is to construct an objective quantitative measure of impulsivity, which serves to make cross-species comparisons, allowing the possibility of translational research. The advantages of these particular protocols include their easy set-up and application, which stems from the relatively small amount of equipment needed and the automated nature of these protocols.

We present three protocols that assess different forms of impulsivity in rats and other small mammals. Intertemporal choice procedures evaluate the tendency to discount the value of delayed outcomes. Differential reinforcement of low rates and feature-negative discrimination evaluate response inhibition capacity with and without punishment for inappropriate responses, respectively.

评估大鼠冲动不同表现的三个实验室程序

The present article provides a guide for the conduction and analysis of three conditioning-based protocols to evaluate impulsivity in rats. Impulsivity is ...