我们要感谢马修 F S 拉什沃思博士 (牛津大学实验心理学系) 和 Sakagami 博士正道 (脑科学研究中心, 玉川大学), 他们对项目的启动和细节的宝贵意见的协议。我们感谢百合博士的批评和编辑手稿。该项目由山本理奖励研究项目 (100226201701100443) 支持 q. z, 脑科学项目, 新科学倡议中心, 国立自然科学研究院 (BS291003) 到 q. Z, 山本理老化项目 (10026-201701100263-340120) 到 Q Z 和 jsp Kakenhi 资助为年轻科学家 (B) (17841749) 到 Q.Z。
作者贡献: Q. Z 构思并启动该项目, 进行了实验和数据分析, 协调了实验室与奥哈拉 & 有限公司的工作, q. z 和 Y 写的手稿, S. 我监督了这个项目。

所有实验性的协议都由山本理脑科学研究所的动物护理和使用委员会批准。
1. 动物制剂
<ol><li>根据实验目的, 选择实验小鼠 (或大鼠) 的性别、年龄、基因型和药理治疗。 注: 在这里我们展示了四只雄性 C57B/6 小鼠2月大的表现。</li>
	<li>把老鼠放在标准条件下的房间里 (12 小时 light/12 h 黑暗循环, 上午8:00 至下午8:00 之间的灯, 22 1°C)。 注: 如果目的是比较两种基因型之间的差异, 每笼4小鼠, 并包括每个基因型的2小鼠。</li>
	<li>处理鼠标2分钟/天5天, 使他们熟悉人类接触。喂养他们与测量的口粮, 使他们的体重大致维持在80–85% 的自由喂养重量在整个实验。提供水上广告随意。</li>
	<li>习惯老鼠到实验室, 把所有老鼠从老鼠屋里转移到实验室, 每天实验前30分钟。</li>
	<li>每天在同一时间开始实验, 以避免昼夜节律对动物性能的影响。</li>
</ol>2. 动物适应迷宫
<ol><li>与鼠标处理同时开始适应迷宫 (2 分钟/天)。在这个阶段保持所有的门打开。在5天内进行习惯性的适应。</li>
	<li>1天, 在迷宫中分散食物颗粒。</li>
	<li>在2和3天, 将小球分散在两个目标臂上。</li>
	<li>在4和5天, 把球团只在两个目标框。</li>
	<li>每天, 在放置小球后, 将小鼠放在 T 迷宫的起始箱中四组, 并允许老鼠探索迷宫10分钟。 注: Habituating 四组的老鼠会帮助他们互相学习, 加快训练。</li>
</ol>3. 对上帝抵抗军的 HRA 的动物歧视
注意: 此协议包括基于延迟和基于工作的决策测试。然而, 根据目的, 研究人员只能测试其中之一, 或两者。控制软件 (材料表) 用于在以下步骤中自动控制 T 迷宫设置。如果要对基于努力的决策进行测试, 在强行进入阶段给 HRA 和上帝抵抗军带来障碍。然后动物将被训练为歧视和障碍攀登同时。饥饿的老鼠积极攀登障碍, 在这个阶段之后, 他们都能熟练地攀爬。因此, 没有必要从较低的屏障开始使用此协议。
<ol><li>
强制臂进入阶段
	<ol>
<li>打开控制软件的参数注册窗口, 然后按如下方式设置参数 (图 3)。<ol>
<li>选择 "阶段选项"。将 "试用号" 设置为 10, 以便每只动物每天进行10次试验, 持续5天。 注: 在他/她自己的实验中, 你可以选择一个不同的试验编号。</li>
			<li>将 "持续时间" 设置为 900s, 以便每天对一只鼠标的训练不会超过 900s. 将 "默认启动延迟时间" 设置为3秒, 以便在开始区域检测到动物后, 启动门将打开3秒。</li>
			<li>为 hra 和上帝抵抗军设置 "颗粒数", 以便在 hra 中总自动分配4个小球, 并在上帝抵抗军中配发1粒。 注: 在我们的实验中, 我们发现1:4 是最佳的比率时, 10 毫克糖丸使用。如果我们使用6到10粒, 老鼠不能吃完所有的, 并会有遗漏发生。</li>
			<li>将 "延迟时间" 设置为0秒, 以便在这一阶段不会延迟 HRA 和上帝抵抗军。</li>
		</ol>
</li>
		<li>打开控制软件的 "ID 注册" 窗口。根据 HRA 的位置, 无论是左侧还是右侧, 将每个鼠标的 ID 注册到软件。(图 4)。 注意: 位置应与基因型组相对应。对于每个基因组中的 50%, HRA 总是在左边, 而上帝抵抗军总是在右边。对于其他 50%, HRA 总是在右边, 上帝抵抗军总是在左边。</li>
		<li>打开软件的应用程序窗口, 从下拉 "任务" 列表中选择 "决策"。输入主题 ID 并从 "阶段" 下拉列表中选择 "阶段 2"。从 "日" 下拉列表中选择日编号。按 "确定" 按钮进入实验界面窗口。</li>
		<li>在实验界面窗口中, 按 "GetBG" 来注册迷宫的背景信息, 以便在不考虑环境背景的情况下准确跟踪动物。按 "会话开始" 按钮 (图 5)。</li>
		<li>将鼠标放在 "开始" 框中, 然后按遥控器的 "开始" 按钮开始训练。<ol>
<li>注意, 启动门, 一个连接门 (左或右) 将自动打开后3秒;一旦鼠标进入连接区, 起始门就会自动关闭。</li>
			<li>观察, 一旦鼠标进入延迟区域 (无论是左或右), 连接门将自动关闭, 目标门将自动打开。</li>
			<li>观察到一旦鼠标拿起小球, 后门和预启动门就会自动打开。一旦鼠标进入后区, 后门就会自动关闭。</li>
			<li>请注意, 一旦鼠标进入 "开始" 框, 预启动门将自动关闭, 新的试用将开始。 注: 在本训练阶段每天10次的试验中, 该软件将自动确保每只老鼠访问 HRA 5 试验和上帝抵抗军5项试验。</li>
		</ol>
</li>
		<li>每天彻底清扫迷宫。</li>
	</ol>
</li>
	<li>
自由臂进入阶段
	<ol>
<li>注册参数和主题 ID 的方式与在强制进入阶段 (步骤3.1.1 和 3.1.2) 中完成的方法相同。选择"阶段选项"。将 "试用号" 设置为 20, 使每只动物每天进行20次试验, 每日持续7天。</li>
		<li>在 "应用程序" 窗口中, 从 "阶段" 下拉列表中选择 "阶段 3"。根据步骤3.1.3 设置其他参数。</li>
		<li>在 "实验界面" 窗口中, 将 "成功率" 值设置为 80%, 以使训练自动继续, 直到鼠标在80% 的试验中选择 HRA, 或者当鼠标每天完成20次试验 (如在参数设置中注册)。按步骤3.1.4 应用其他操作。</li>
		<li>允许鼠标自由选择一只手臂, 无论是 HRA 还是上帝抵抗军。<ol>
<li>注意, 起始门、两个接线门将在3秒后自动打开;一旦鼠标进入连接区, 起始门就会自动关闭。</li>
			<li>观察, 一旦鼠标选择一只手臂进入延迟区域 (无论是左或右), 连接门将自动关闭, 目标门将自动打开。</li>
			<li>观察到一旦鼠标拿起小球, 后门和预启动门就会自动打开。一旦鼠标进入后区, 后门就会自动关闭。</li>
			<li>请注意, 一旦鼠标进入 "开始" 框, 预启动门将自动关闭, 新的试用将开始。</li>
		</ol>
</li>
	</ol>
</li>
</ol>4. 基于延迟的决策测试
<ol><li>注册参数和主题 ID 与在自由臂进入阶段 (步骤 3.2.1) 中完成的方法相同。将 "延迟时间" 设置为5、1015天1、2天和3天, 以便在5天的 hra 上有1秒的延迟, 十年代延迟为 hra 的 2, 十五年代为 hra 的延迟为3天。</li>
	<li>在 "应用程序" 窗口中, 从 "阶段" 下拉列表中选择 "阶段 4"。以与3.2.2 相同的方式设置其他参数。</li>
	<li>在 "实验界面" 窗口中, 按步骤3.2.3 应用所有操作。</li>
	<li>
允许鼠标自由选择一只手臂, 无论是 HRA 还是上帝抵抗军。
	<ol>
<li>注意, 起始门、两个接线门将在3秒后自动打开;一旦鼠标进入连接区, 起始门就会自动关闭。</li>
		<li>观察到一旦鼠标选择一只手臂进入延迟区域 (无论是左还是右), 接合门就会自动关闭。 注: 如果鼠标选择上帝抵抗军, 球门门将立即自动打开。但是, 如果鼠标选择 HRA, 目标门将在5秒、十年代和十五年代之后自动打开, 分别在1、2、3天。</li>
		<li>观察到一旦鼠标拿起小球, 后门和预启动门就会自动打开。一旦鼠标进入后区, 后门就会自动关闭。</li>
		<li>请注意, 一旦鼠标进入 "开始" 框, 预启动门将自动关闭, 新的试用将开始。 注意: 在这里, 我们训练的老鼠5–7天的每一个延迟条件。然而, 根据我们对转基因或突变小鼠多行检测的经验, 1 天 (20 项试验) 绝对足以看出不同基因型小鼠之间的差异, 并且没有意义延长训练时间 (见图 6为一个例子)。因此, 目前我们只适用于每一个延迟时间1天, 它的工作很好。如果研究人员想根据自己的目的延长训练日, 就没有问题了。</li>
	</ol>
</li>
	<li>可选: 执行与 HRA 反转的测试。要测试鼠标的选择是否是方向首选项的结果, 请切换 HRA 和上帝抵抗军的左/右位置 (可以由软件自动完成), 并允许鼠标自由选择一只手臂, 如4.4。</li>
	<li>可选: 执行延迟控制测试。测试是否有任何赤字是由于空间记忆或奖赏敏感性的改变, 而不是决策的变化造成的结果, 给上帝抵抗军和 HRA 引入十五年代的延迟, 并允许老鼠自由选择一只手臂, 就像在4.4。</li>
</ol>5. 基于努力的决策测试
<ol><li>将屏障引入到 HRA 中, 如图所示 (图 1)。</li>
	<li>设置所有的参数, 并应用所有的操作, 按步骤 3.2-自由臂进入阶段, 并测试动物连续3天。</li>
	<li>允许老鼠自由选择一只手臂, 无论是 HRA 还是上帝抵抗军。 注意: 在这里, 我们训练了老鼠14天。然而, 根据我们对转基因或突变小鼠多行检测的经验, 3 天绝对足以看出不同基因型小鼠之间的差异, 而且没有意义延长训练时间 (见图 6为例).因此, 目前我们只应用3天的努力为基础的测试, 它的工作很好。如果研究人员想根据自己的目的延长训练日, 就没有问题了。</li>
	<li>可选: 执行与 HRA 反转的测试。要测试鼠标的选择是否是方向首选项的结果, 请切换 HRA 和上帝抵抗军的左/右位置 (可以由软件自动完成), 并允许鼠标自由选择一只手臂, 如步骤5.3 所做的那样。</li>
	<li>可选: 执行努力控制测试。测试是否有任何赤字是由于空间记忆或奖赏敏感性的改变, 而不是决策的变化所造成的, 给上帝抵抗军和 HRA 带来障碍, 并允许老鼠自由地选择一只手臂, 就像在步骤5.3 中一样。</li>
</ol>6. 数据分析
<ol><li>
直接从控制软件获取数据和结果。
	<ol>
<li>请注意, 该软件将自动记录实验日期、开始和完成时间、工期、试用编号、hra 的位置、hra 和上帝抵抗军中的颗粒数、位置 (X、Y) 和移动跟踪等,这些数据文件夹中的每个鼠标.</li>
		<li>检查软件是否自动分析下列项目, 并在每个动物 ID 下的 "结果" 文件夹中记录它们: 持续时间、试用编号、hra 选择号、上帝抵抗军选择号码、hra 选择百分比、上帝抵抗军选择百分比、总移动距离、和总连接时间。</li>
	</ol>
</li>
	<li>对来自所有实验的数据进行统计分析 (分块变异因子), 以天/期为内标因子和群体因素 (基因组或不同实验条件的群体) 作为主题因子。</li>
	<li>分析了群体因素的主要影响, 如果白天/会话与群体因素之间没有交互作用。如果日/会话与组因子之间有显著的交互作用, 则应用后向比较。</li>
</ol>

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作者没有什么可透露的。

决策是在进化15中高度保守的认知过程。人类和动物可以评估相对于潜在报酬的竞争行动选择的成本, 然后做出选择。患有多种神经系统疾病和心理疾病的患者表现出16不同形式的决策赤字。因此, 重要的是要调查的神经生物学和病理生理机制的基础上的决策过程。在过去的几年中, 基于延迟和努力的决策正在吸引越来越多的研究兴趣。此外, 啮齿类动物, 特别是大鼠已广泛用于研究这两种形式的决策17。
许多研究导致了有趣的发现使用一个行为任务涉及一个 T 迷宫设备与 HRA 和上帝抵抗军2,6,7,8,9,10,18,19,20,21,22. 在任务中, HRA 将大量奖励与时间延迟或努力劳累联系在一起。在上帝抵抗军, 动物可以立即获得一个小的奖励, 没有任何时间延迟和体力的努力。传统的方法依赖于人类实验者的人工干预。在每次试验中, 实验者都需要数出小球并将它们放在 hra 和上帝抵抗军的食物托盘中, 将目标门放在 hra 和上帝抵抗军上, 然后将动物放在开始臂的末端。当动物进入任何一个武器, 需要放置一个连接门, 以限制动物的目标手臂。根据协议, 实验者需要计算时间并在设定的延迟后打开目标门。在动物进入目标区并获得球团后, 实验者需要将其返回笼子, 记录动物的手臂选择和行为。然后实验者需要为下一次试验准备 T 迷宫门和小球。整个训练和测试过程是极大的时间和劳力密集。此外, 在不同的实验室中缺乏标准化是另一个值得关注的问题。
本文提出了一种基于视频跟踪系统 (图 7) 的改进的自动 T 迷宫装置的协议, 以解决传统协议的问题。通过引入一个 "后门" 和 "后廊" 到传统的 T 迷宫, 我们得到了一个 "一分为二等腰三角形三角形" 形状的迷宫。此设置的优点是 (1) 全面自动化的行为训练和测试。这就消除了实验者主体性的影响, 最大限度地减少了人类的时间和劳动承诺。我们在实验室有四个设置, 这样四只老鼠可以由一个实验者同时训练或测试, 这是不可能用传统的协议来完成的。(2) 软件具有灵活性, 因为控制软件允许实验者自由地设置多个参数, 包括颗粒数、延迟时间、门开和闭合、试数、持续时间和跟踪模式。因此, 该系统可以满足不同类型的实验需要。(3) 有广泛的兼容性, 因为 T 迷宫的所有滑动门都被设计成在迷宫的基础上储存时, 它们是开放的。因此, 该装置可以很容易地集成到不同的生理系统, 包括 optogenetic/光操作,在体内电生理记录和微透析。此外, 为了排除小鼠由于位置偏好而选择 HRA 的可能性, 我们建议对延迟和努力为基础的检测应用一个控制测试。通过均衡两个目标武器的成本, 动物们有机会以同样的成本体验两种奖励结果。选择可以简单地根据奖励差异, 从而消除了在决定之前兼顾成本和效益的需要。这也检验了动物的选择是否有任何改变是由于无法扩大成本或报酬, 或记忆力不足, 而不是他们评估决定的方式发生了变化。
在我们的实验室, 我们已经分析了大约10株小鼠与此设置。有一个例子显示在代表性数据中, 缰: DTA 小鼠在延迟和基于努力的决策中表现出强健的表型。即, 在缰的时间和努力中, 奖励价值被强烈的贴现: DTA 小鼠。结果揭示了缰在冲动控制中的重要作用。此外, 我们在决策过程中对自由移动小鼠应用了硅探针记录 (未发布的数据)。所有的实验都为自动化安装的能力提供了验证基准。因此, 基于 T 迷宫的自动化设备决策标准化协议适用于检测遗传效应、药理效应和神经回路对啮齿动物延迟和努力贴现的影响。总之, 该设置具有许多优点, 可以作为基于延迟和努力的决策检测的理想系统。

人类和其他动物评估成本 (包括延迟, 努力和风险) 得到奖励, 然后作出决定选择某种行动的过程。决策赤字出现在许多精神疾病, 包括精神分裂症 (深圳), 注意缺陷多动障碍 (ADHD), 强迫症 (强迫), 帕金森病 (PD) 和成瘾1。对人类和猴子的研究显示, 几个关键的大脑区域参与决策2,3,4。虽然灵长类动物从事更复杂的决策, 但据报道, 啮齿类动物能够做出自适应的决定, 在一个奖励意外变化频繁的环境中生存。此外, 由于 chemogenetic 工具、optogenetic 工具和基因工程小鼠的可用性, 可以对小鼠模型中的神经回路机制和分子机制进行深入研究。在评价啮齿动物决策行为时, 有多种任务, 包括注意设置移位任务、刻苦或延迟型 T 迷宫任务、爱荷华州赌博任务、视觉识别反转学习任务5等。类似的 T 迷宫成本效益协议最初是由皮埃尔集团6开发的, 并被用来检查两种类型的决策成本 (延迟和努力) 对自由移动啮齿目动物7,8的影响, 9,10。这项任务的特殊好处是, 动物不需要被训练, 以按杠杆或挖碗。相反, 动物在一只手臂 (HRA) 中选择高奖赏高成本选项, 或者在另一只手臂 (上帝抵抗军) 中进行低报酬低成本选择。因此, 此任务更易于执行。
在基于延迟的范式中, 一旦实验动物进入目标臂, 就会引入一个结门, 使动物保持在目标臂中。如果动物选择了上帝抵抗军, 上帝抵抗军的目标门立即被收回, 少量的食物交付。如果动物选择 hra, hra 上的目标门在所需的延迟后被收回, 并且大量的食物颗粒被交付 (图 1A)。在基于努力的范式中, HRA 被屏障阻挡, 动物必须爬过它才能获得大量的小球 (图 1B)。一般而言, 基于延迟的范式对于测试动物模型的冲动性非常有用, 并且基于努力的方法可以帮助找出冷漠的动物2,4,11,12,13. 迄今为止, 研究人员一直在进行这种化验, 方法是手动计算延时、插入和撤门、操纵努力屏障、计算颗粒数、将小球放入位置、放置和返回动物。, 记录动物对每项试验的选择。这些劳动和时间成本给研究人员造成了严重的实验瓶颈, 阻碍了这种行为分析的广泛应用。在目前的工作中, 我们开发了一个基于 T 迷宫的设置来评估啮齿类动物的延迟或基于努力的决策, 具有完全自动化、标准化和高通量能力。
装置
与商业制造商 (见材料表) 合作, 我们开发了一种使用基于软件的仪器控制的改进的自动 T 迷宫设备 (图 2)。特别是, 我们引入了一个 "后门" 和 "后退方式" 相比, 传统的 T 迷宫 (图 2), 使动物可以回到起点自己和开始一个新的试验。T 型迷宫为哑光灰色, 当实验条件和软件设置得当时, 可以检测出黑白小鼠。它由三胳膊组成: 一开始胳膊和二个目标胳膊, 每410毫米长度以 V 形墙壁155毫米在高度, 一个30毫米宽度的基地和一个开放顶部155毫米宽度。V 形走廊可以有效地防止老鼠跳跃。此外, V 型走廊更容易在体内使用电缆进行录音。开始框连接到开始臂的末尾。目标框附加到每个目标臂的末尾。每个目标框中都安装了自动食品分配器, 以提供预定义数量的甜食品颗粒。通过红外传感器检测颗粒进气量, 并由计算机自动记录。每个目标框都通过直线通道连接到 "开始" 框。动物一旦完成试验, 就可以通过走廊自主返回到起始箱。有滑动门在高度155毫米在起点和目标箱子的入口和出口。另外, 滑动门位于每个目标臂的入口处, 以防止动物在做出选择后向后移动 (图 2A)。所有滑动门均由计算机控制, 可自动开启和关闭。一个高灵敏度 1/2 "电荷耦合器件 (CCD) 戴高相机设置在仪器之上, 以跟踪动物的行为。透镜的焦距为2.8–12毫米。相机的位置大约是1.9 米高。由于迷宫的高度是 0.5 m 从地板, 照相机和迷宫之间的距离是大约 1.4 m (图 2B)。从 CCD 相机获得的跟踪数据被用来控制 T 迷宫, 当动物进入某些感兴趣的区域时, 打开和关闭特定的门 (ROIs)。用于基于努力的范式的障碍是三维直角三角形的形状 (图 2C), 它完全适合于 V 形墙体, 身高约为155毫米。动物必须缩放垂直一侧, 但能够下降45°斜坡。在实验中, 该仪器在100勒克斯照明。实验中使用的糖丸 (见材料表) 和硅胶 (见材料表) 用于保持颗粒干燥。

<table><tbody><tr><td>automated t-maze for decion making testing</td><td>O&amp;rsquo;HARA &amp;amp; Co.,ltd</td><td>no catalog number, customorized</td><td>Address requested by the reviewer: 4-28-16 Ekoda, Nakano-ku, Tokyo 165-0022&lt;br/&gt; TEL: 81-3-3389-2451 FAX:81-3-3389-2453</td></tr><tr><td>slica gel&amp;nbsp;</td><td>Nacalai Tesque</td><td>1709155</td><td></td></tr><tr><td>AIN-76A Rodent Tablet 10mg</td><td>Test Diet</td><td>1811213(5TUL)</td><td>Manufactured for Japan,SLC</td></tr><tr><td>Time TM1 software&amp;nbsp;</td><td>O&amp;rsquo;HARA &amp;amp; Co.,ltd</td><td>no catalog number</td><td></td></tr><tr><td>SPSS statistics V21.0</td><td>IBM</td><td></td><td></td></tr></tbody></table>

an automated t-maze based apparatus and protocol for analyzing delay- and effort-based decision making in  free moving rodents

许多神经和精神病患者在决策中表现出困难和/或缺陷。啮齿类动物模型有助于加深对决策问题背后的神经生物学原因的理解。基于成本效益的 T 迷宫任务用于测量决策, 在这种情况下, 啮齿动物可以在高奖赏臂 (HRA) 和低奖赏臂 (上帝军) 之间进行选择。T 迷宫决策任务有两种范式, 即成本是时间延迟, 另一种是体力工作。这两种范式都需要对实验动物、多门、球团奖励和手臂选择记录进行繁琐和劳动密集型的管理。在目前的工作中, 我们发明了一种基于传统的 T 形迷宫的设备, 用于颗粒输送, 门管理和选择记录的全自动。这种自动设置可用于评估啮齿类动物的延迟和基于努力的决策。通过本文所述的协议, 我们的实验室研究了多基因修饰小鼠的决策表型。在代表性数据中, 我们发现, 消融内侧缰的小鼠表现出厌恶的延迟和努力, 并倾向于选择立即和毫不费力的奖励。该协议有助于减少实验者干预引起的变异性, 提高试验效率。此外, 使用此处描述的设置, 可以很容易地在决策任务中应用慢性硅探针或微电极记录、光纤成像和/或操作神经活动。

图 6显示了缰消融小鼠 (缰: DTA 小鼠)14与野生型 littermate 控制小鼠 (CT 小鼠) 进行延迟和努力的决策任务的例子。两个缰: DTA 小鼠和两个 CT 小鼠在断奶后在一个笼子里共同居住。
在基于延迟的决策测试中 (图 6A), 在任何阶段, 基因型和会话之间没有显著的交互作用, 包括歧视训练阶段 (HRA 延迟时间为 0) 和基于延迟的决策测试阶段 (当 HRA 的延迟时间分别为5秒、十年代和十五年代) 时。当延迟时间为5时, 基因型的主要作用不显著。然而, 当延迟时间延长到十年代和十五年代, 缰: DTA 小鼠显示, 与 CT 小鼠相比, HRA 访问的百分比显著降低。这些结果表明, 缰的消融降低了小鼠的偏爱, 等待更大的奖赏, 相反, 当等待时间是10秒甚至更长时, 就会立即选择一个小的奖赏。数据表明, 缰可能是控制冲动和/或时间成本/效益评估的一个重要的大脑结构, 使动物更容易容忍延迟获得大奖励。
在基于努力的决策测试 (图 6B) 中, 缰中 hra 访问的百分比显著降低: DTA 小鼠在 hra 中放置屏障时, 不管 hra 的左/右定位 (1x 屏障相位和反转相位)).这意味着缰的表型: DTA 小鼠不是由于空间偏好和记忆的缺陷。在努力控制试验中, 在两个目标武器 (2×障碍阶段) 都设置了障碍, 而上帝抵抗军和 HRA 都与高的努力联系在一起。因此, 对于选择低奖赏或高奖赏的动物来说, 付出的代价是相同的。缰: DTA 小鼠比上帝抵抗军更频繁地访问了 hra, 并在最后一届会议上达到了可比的 hra 访问号码 (会议 5)。这一结果表明, 缰的奖励敏感性和空间记忆: DTA 小鼠是完整的。数据阐明, 缰可能在努力成本/效益评估中发挥重要作用, 允许动物投入更多的工作获得更大的回报。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/57895/57895fig1.jpg"> 图 1: 传统的用于决策测试的 T 迷宫设备示意图.(A) 基于延迟的决策测试装置。动物被放置在开始臂, 并选择两个目标武器, HRA 和上帝抵抗军。当动物选择了 HRA, 他们必须等待 (取决于延迟时间秒) 得到一个更大的奖励。研究人员需要手动处理每项试验的动物、小球和门。(B) 基于努力的决策测试装置。动物被放置在开始臂, 并选择两个目标武器, HRA 和上帝抵抗军。当动物选择了 HRA 时, 他们不得不爬过一个三角形的屏障来获得更大的奖赏。研究人员需要手工处理动物, 小球, 门和障碍的每一个试验。<a href="https://www.jove.com/files/ftp_upload/57895/57895fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/57895/57895fig2.jpg"> 图 2: 用于决策测试的自动化 T 迷宫设置.(A) 自动安装的顶级视图。(B) 自动设置的侧面视图。(C) 3D 直角三角形屏障, 用于基于努力的决策测试, 从左到右, 分别是侧面视图、相对侧面视图和斜边侧面视图。在商业制造商的许可下编辑的原始技术照片。GBL: 目标箱子 (左), 英国: 目标箱子 (正确), GDL: 目标门 (左), GDR: 目标门 (右), DAL: 延迟区域 (左), DAR: 延迟区域 (右), JDL: 接线门 (左), JDR: 接线门 (右), BDL: 后门 (左), BDR: 后门 (右), CCD: 电荷耦合设备摄像头)。<a href="https://www.jove.com/files/ftp_upload/57895/57895fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/57895/57895fig3v2.jpg"> 图 3: 参数注册窗口.<a href="https://www.jove.com/files/ftp_upload/57895/57895fig3v2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/57895/57895fig4v2.jpg"> 图 4: 主题 ID 注册窗口.<a href="https://www.jove.com/files/ftp_upload/57895/57895fig4v2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/57895/57895fig5v2.jpg"> 图 5: 实验界面窗口.<a href="https://www.jove.com/files/ftp_upload/57895/57895fig5v2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/57895/57895fig6v2.jpg"> 图 6: 缰中基于延迟和基于努力的决策: DTA 小鼠.(A) 缰的基于延迟的决策测试: DTA 小鼠 (小鼠12–14月大, n = 8/基因型)。当延迟时间为0和5秒时, HRA 选择的百分比与基因型比较, 但缰: DTA 小鼠的延迟时间为十年代和十五年代。当延迟时间为5秒时, genotype×day 相互作用: F (114) = 0.594, p = 0.236;基因型的影响: F (114) = 0.61, p = 0.45;当延迟时间是十年代: genotype×day 相互作用: F (114) = 37.5, p = 0.346;基因型的影响: F (114) = 32.4, p < 0.0001;当延迟时间是十五年代: F (114) = 38.7, p = 0.243;基因型的影响: F (114) = 31.6, p ≤0.0001。(B) 缰的基于努力的决策测试: DTA 小鼠 (小鼠12-14 月大, n = 9/基因型)。在1x 屏障阶段, 基因型与会话之间有显著的交互作用 (genotype×day 相互作用: F (116) = 2.11, p = 0.015),后组配对比较显示缰的 HRA%: DTA 小鼠显著下降会话。在逆转阶段, 基因型与会话之间没有显著的交互作用 (genotype×day 相互作用: F (116) = 1.61, p = 0.08)。缰: DTA 小鼠对 HRA 的访问量明显低于 CT 小鼠 (主要基因型效应: F (116) = 8.18, p = 0.01)。在2×barriers 阶段, 基因型和会话之间存在着显著的交互作用, 在3和会话 4 (2x 屏障阶段: genotype×day 交互作用: F (116) = 3.9, p = 0.0067) 中有显著差异。缰: DTA 小鼠在最后一届会议5达到了与 CT 小鼠相比的 HRA 访问数字。数据代表的是 "电子扫描电镜" ** < 0.01;p < 0.001。<a href="https://www.jove.com/files/ftp_upload/57895/57895fig6v2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/57895/57895fig7v2.jpg"> 图 7: 决策测试流程图 (延迟或基于工作量)。

Watch this Scientific Journal Video about An Automated T-maze Based Apparatus and Protocol for Analyzing Delay- and Effort-based Decision Making in  Free Moving Rodents at JoVE.com

An Automated T-maze Based Apparatus and Protocol for Analyzing Delay- and Effort-based Decision Making in  Free Moving Rodents

本文介绍了我们发明的一种自动化的 T 迷宫装置, 并在此基础上对自由移动啮齿动物的基于时延的决策和基于努力的决策进行了分析。

基于自动 T 迷宫的自由移动啮齿动物延迟和努力决策分析装置及协议

Many neurological and psychiatric patients demonstrate difficulties and/or deficits in decision making. Rodent models are helpful to produce a deeper ...

an-automated-t-maze-based-apparatus-protocol-for-analyzing-delay

Research

JoVE Journal

Behavior

13.5K 次观看.  RIKEN. 本文介绍了我们发明的一种自动化的 T 迷宫装置, 并在此基础上对自由移动啮齿动物的基于时延的决策和基于努力的决策进行了分析。

视频: An Automated T-maze Based Apparatus and Protocol for Analyzing Delay- and Effort-based Decision Making in  Free Moving Rodents

A Fully Automated Rodent Conditioning Protocol for Sensorimotor Integration and Cognitive Control Experiments

Rodents have been traditionally used as a standard animal model in laboratory experiments involving a myriad of sensory, cognitive, and motor tasks. Higher cognitive functions that require precise control over sensorimotor responses such as decision-making and attentional modulation, however, are typically assessed in nonhuman primates. Despite the richness of primate behavior that allows multiple variants of these functions to be studied, the rodent model remains an attractive, cost-effective alternative to primate models. Furthermore, the ability to fully automate operant conditioning in rodents adds unique advantages over the labor intensive training of nonhuman primates while studying a broad range of these complex functions.
Here, we introduce a protocol for operantly conditioning rats on performing working memory tasks. During critical epochs of the task, the protocol ensures that the animal's overt movement is minimized by requiring the animal to 'fixate' until a Go cue is delivered, akin to nonhuman primate experimental design. A simple two alternative forced choice task is implemented to demonstrate the performance. We discuss the application of this paradigm to other tasks.

A fully automated protocol for rodent operant conditioning is proposed. The protocol relies on precise temporal control of behavioral events to investigate the extent to which this control influences neural activity underlying sensorimotor integration and cognitive control experiments.

为感觉运动整合与认知控制实验完全自动化的啮齿动物调理协议

Rodents have been traditionally used as a standard animal model in laboratory experiments involving a myriad of sensory, cognitive, and motor tasks. ...

科研

行为学

Barnes Maze Testing Strategies with Small and Large Rodent Models

Spatial learning and memory of laboratory rodents is often assessed via navigational ability in mazes, most popular of which are the water and dry-land (Barnes) mazes. Improved performance over sessions or trials is thought to reflect learning and memory of the escape cage/platform location. Considered less stressful than water mazes, the Barnes maze is a relatively simple design of a circular platform top with several holes equally spaced around the perimeter edge. All but one of the holes are false-bottomed or blind-ending, while one leads to an escape cage. Mildly aversive stimuli (e.g.&#160;bright overhead lights) provide motivation to locate the escape cage. Latency to locate the escape cage can be measured during the session; however, additional endpoints typically require video recording. From those video recordings, use of automated tracking software can generate a variety of endpoints that are similar to those produced in water mazes (e.g.&#160;distance traveled, velocity/speed, time spent in the correct quadrant, time spent moving/resting, and confirmation of latency). Type of search strategy (i.e.&#160;random, serial, or direct) can be categorized as well. Barnes maze construction and testing methodologies can differ for small rodents, such as mice, and large rodents, such as rats. For example, while extra-maze cues are effective for rats, smaller wild rodents may require intra-maze cues with a visual barrier around the maze. Appropriate stimuli must be identified which motivate the rodent to locate the escape cage. Both Barnes and water mazes can be time consuming as 4-7 test trials are typically required to detect improved learning and memory performance (e.g.&#160;shorter latencies or path lengths to locate the escape platform or cage) and/or differences between experimental groups. Even so, the Barnes maze is a widely employed behavioral assessment measuring spatial navigational abilities and their potential disruption by genetic, neurobehavioral manipulations, or drug/ toxicant exposure.

The dry-land Barnes maze is widely used to measure spatial navigation ability in response to mildly aversive stimuli. Over consecutive days, performance (e.g.&#160;latency to locate escape cage) of control subjects improves, indicative of normal learning and memory. Differences between rats and mice necessitate apparatus and methodology changes that are detailed here.

巴恩斯迷宫测试策略与小型和大型啮齿类动物模型

Spatial learning and memory of laboratory rodents is often assessed via navigational ability in mazes, most popular of which are the water and dry-land ...

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 ...

The Double-H Maze: A Robust Behavioral Test for Learning and Memory in Rodents

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their behavioral flexibility and required memory duration, the number of goals and pathways, and also the overall task complexity. A confounding feature in many of these tasks is the lack of control over the strategy employed by the rodents to reach the goal, e.g., allocentric (declarative-like) or egocentric (procedural) based strategies. The double-H maze is a novel water-escape memory task that addresses this issue, by allowing the experimenter to direct the type of strategy learned during the training period. The double-H maze is a transparent device, which consists of a central alleyway with three arms protruding on both sides, along with an escape platform submerged at the extremity of one of these arms.
Rats can be trained using an allocentric strategy by alternating the start position in the maze in an unpredictable manner (see protocol 1; &#167;4.7), thus requiring them to learn the location of the platform based on the available allothetic cues. Alternatively, an egocentric learning strategy (protocol 2; &#167;4.8) can be employed by releasing the rats from the same position during each trial, until they learn the procedural pattern required to reach the goal. This task has been proven to allow for the formation of stable memory traces.
Memory can be probed following the training period in a misleading probe trial, in which the starting position for the rats alternates. Following an egocentric learning paradigm, rats typically resort to an allocentric-based strategy, but only when their initial view on the extra-maze cues differs markedly from their original position. This task is ideally suited to explore the effects of drugs/perturbations on allocentric/egocentric memory performance, as well as the interactions between these two memory systems.

The goal of this protocol is to investigate spatial cognition in rodents. The double-H water maze is a novel test, which is particularly useful to elucidate the different components of learning, consolidation and memory, as well as the interplay of memory systems.

双-H迷宫:一个强大的行为测试学习和记忆的鼠害

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their ...

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

A Fully Automated and Highly Versatile System for Testing Multi-cognitive Functions and Recording Neuronal Activities in Rodents

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be investigated in a single task sequence. The unique feature of this system is a custom-made, acoustically transparent chamber that eliminates many of the issues associated with auditory cue control in most commercially available chambers. The ease with which operant devices can be added or replaced makes this system quite versatile, allowing for the implementation of a variety of auditory, visual, and olfactory behavioral tasks. Automation of the system allows fine temporal (10 ms) control and precise time-stamping of each event in a predesigned behavioral sequence. When combined with a multi-channel electrophysiology recording system, multiple cognitive brain functions, such as motivation, attention, decision-making, patience, and rewards, can be examined sequentially or independently.

In this report, we present a fully automated and highly versatile system capable of simultaneously testing multi-cognitive behaviors and recording neuronal activities for rodents.

一个多认知功能测试，并记录在啮齿动物的神经元活动的完全自动化和高度灵活的系统

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be ...

A Behavioral Assay for Investigating the Role of Spatial Memory During Instinctive Defense in Mice

Evolution has selected a repertoire of defensive behaviors that are essential for survival across all animal species. These behaviors are often stereotyped actions elicited in response to innately aversive sensory stimuli, but their success requires enough flexibility for adapting to different spatial environments, which can change rapidly. Here, we describe a behavioral assay to evaluate the influence of learned spatial knowledge on defensive behaviors in mice. We have adapted the widely used Barnes maze spatial memory assay to investigate how mice navigate to a shelter during escape responses to innately aversive sensory stimuli in a novel environment, and how they adapt to acute changes in the environment. This new assay is an ethological paradigm that does not require training and exploits the natural exploration patterns and navigation strategies in mice. We propose that the set of protocols described here are a powerful means of studying goal-directed behaviors and stimulus-triggered navigation, which should be of interest to both the fields of instinctive behaviors and spatial memory.

This protocol describes a behavioral assay, based on the Barnes maze test, to study how instinctive defensive actions are modified by the knowledge of spatial environment.

一种研究空间记忆在小鼠本能防御中作用的行为分析法

Evolution has selected a repertoire of defensive behaviors that are essential for survival across all animal species. These behaviors are often ...

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 ...

Utilizing a Reconfigurable Maze System to Enhance the Reproducibility of Spatial Navigation Tests in Rodents

Several maze shapes are used to test spatial navigation performance and behavioral phenotypes. Traditionally, each experiment requires a unique maze shape, thus requiring several separate mazes in different configurations. The maze geometry cannot be reconfigured in a single environment to accommodate scalability and reproducibility. The reconfigurable maze is a unique approach to address the limitations, allowing quick and flexible configurations of maze pathways in a repeatable manner. It consists of interlocking pathways and includes feeders, treadmills, movable walls, and shut-off sensors. The current protocol describes how the reconfigurable maze can replicate existing mazes, including the T-shaped, plus-shaped, W-shaped, and figure-eight mazes. Initially, the T-shaped maze was constructed inside a single experimental room, followed by modifications. The rapid and scalable protocol outlined herein demonstrates the flexibility of the reconfigurable maze, achieved through the addition of components and behavioral training phases in a stepwise manner. The reconfigurable maze systematically and precisely assesses the performance of multiple aspects of spatial navigation behavior.

The present protocol describes a reconfigurable maze, a unique system for testing spatial navigation and behavioral phenotypes in rodents. The adaptability of this maze system enables the execution of various experiments in a single physical environment. The ease of structural rearrangement generates reliable and reproducible experimental results.

利用可重构的迷宫系统提高啮齿动物空间导航测试的可重复性

Several maze shapes are used to test spatial navigation performance and behavioral phenotypes. Traditionally, each experiment requires a unique maze ...

Behavioral Training Procedures for Head-fixed Virtual Reality in Mice

Virtual reality (VR) combined with head-fixation is increasingly being utilized in behavioral neuroscience studies as it allows complex behavioral assays to be performed in head-fixed mice. This enables precise behavioral recordings while incorporating various neurophysiological techniques that require head-fixation to minimize movement-related signal noise during neural recordings. However, despite the growing use of VR, there is little published data on the detailed methodology of how to implement it. In this study, a training protocol is developed whereby male and female C57B16/J mice are trained to run down a virtual linear corridor, the length of which is increased from 1-3 m over multiple training sessions. Building upon this foundation, this study investigated the feasibility of mice performing complex behaviors within VR using a Y-maze paradigm. The task required navigating to the arm with black walls from the choice point in the Y-maze. After reaching a criterion of two consecutive days equal to or greater than 70% correct, the mice progressed to increasingly difficult sensory discrimination. The findings provide important details on the methodologies useful for the successful training of mice in VR and demonstrate that mice exhibit learning capabilities in navigating the Y-maze. The methodology presented not only offers insights into training duration in VR-based assays but also underscores the potential for probing intricate behaviors in mice, opening avenues for more comprehensive neuroscience investigations.

The article describes the experimental procedures for the commonly used linear track virtual reality (VR) paradigm in mice as well as determining the feasibility of running complex VR tasks by testing a Y-shaped signal discrimination task.

Virtual reality (VR) combined with head-fixation is increasingly being utilized in behavioral neuroscience studies as it allows complex behavioral assays ...