我们感谢丽贝卡伯韦尔使用她的实验室在录影录音和罗伯特维拉为帮助与早先版本这本手稿。

这里描述的所有方法都已被普罗维登斯学院的机构动物护理和使用委员会 (IACUC) 批准。
1. 制造测试仪器
<ol><li>使用社会互动室 (76.2 x 114.3 厘米), 分为3个独立的房间。<ol>
<li>将中心房间分成一个样本, 用一个纸板插入器来延迟曲面, 可以通过滑动通过附着在侧面墙上的金属轨道来解除。</li>
	</ol>
</li>
	<li>使用不透明的接触纸覆盖清晰的墙壁, 以防止视觉进入其他房间。</li>
	<li>右侧房间的一侧 (测试室; 见图 1), 用沙子颗粒和红色接触纸盖住地板。这将成为视觉和触觉的歧视提示的下降选项。</li>
	<li>在测试室的另一侧, 用凸起的条纹覆盖地板, 使用带屏蔽胶带作为选择测试的视觉和触觉的判别提示。用黑色接触纸遮住条纹。保持房间的其余中间第三不变。</li>
</ol>2. 制定首选的食品奖励等级/偏好测试
<ol><li>为测试选择可能的食物奖励。确保一个奖励是一个更大的奖励, 因此 "首选" 的其他。<ol>
<li>用一整片麦片作为更大的 "首选" 奖赏和一¼的谷物, 作为小的 "不太喜欢" 的奖励。 注:谷物可以包括甜麦片或脱水棉花糖。它也可以使用两种不同的奖励 (如谷物与标准啮齿动物颗粒 (TestDiet, 45 毫克)), 但很难确定一个可靠的偏好排名内和跨个人时, 质量, 而不是数量是不同奖励的变化来源。</li>
	</ol>
</li>
	<li>在测试开始之前, 把老鼠 (成年男性长埃文斯) 带到黑暗的测试室30分钟。使用红色照明照亮测试室, 如果老鼠是在他们的一天的黑暗阶段的周期。如果没有, 在正常光照条件下测试, 如视频中所示。</li>
	<li>把这两个奖励, 整个谷物和¼一块麦片, 在房间的右室的相对角落 (见图 1)。</li>
	<li>把老鼠放在房间的另一侧, 面对远离的奖赏, 并允许老鼠吃的奖励之一。 注:为了通过偏好测试, 并确保更大的谷物是首选的奖励, 该主题必须吃 4 5 试验 (每届5次试验) 的更大的奖励2个连续的会话。每天运行两个会话。<ol>
<li>如果在10次疗程后, 老鼠没有表现出对大片谷物的偏爱, 那就引入不同的奖赏。</li>
	</ol>
</li>
	<li>在整个测试中, 为了确保老鼠的喜好保持一致, 每周进行一次偏好检查。<ol>
<li>以与首选项测试相同的方式运行首选项检查, 并将整个和1/4 块谷物放在房间的相对角上。 注: 2 项试验包括首选检查。</li>
		<li>如果大鼠选择对两种试验都不太喜欢的奖励, 则在5项试验的完全偏爱试验中试验大鼠。<ol>
<li>如果老鼠通过, 它可能继续测试。</li>
			<li>如果老鼠失败了, 停止测试并引入不同类型或数量的奖励。</li>
		</ol>
</li>
	</ol>
</li>
</ol>3. 一般方法
<ol><li>在测试过程中使用一致的实验, 因此老鼠完全习惯于实验者的气味和处理。 注:据推测, 在试验之前, 老鼠已经被人类至少处理了1周, 因此他们习惯于被实验者拾起和携带。</li>
	<li>在测试开始使他们适应新的房间前30分钟, 把老鼠放在鞋盒笼子里 (家庭笼子) 里。</li>
	<li>如果测试是在大鼠周期的黑暗阶段进行的, 保持测试环境的黑暗, 除了红灯照明设备。</li>
	<li>确保有香味的选择杯按适当的顺序排列, 由包括制衡的随机脚本决定。用整块麦片诱饵正确的杯子。</li>
	<li>只允许大鼠在一杯中挖, 并立即从仪器中取出的老鼠, 一旦一杯已被选中, 通过挖掘表明。把老鼠放在家里的笼子里, 老鼠可以在那里吃完赏金。</li>
	<li>试验间隔时间固定为4分钟, 一旦老鼠在笼子里, 就开始计时秒表。<ol>
<li>在使用过程中, 用70% 异丙醇清除所有溢出的沙子并擦拭地板, 清洁设备。如果在上一次试验中没有被吃掉, 就放弃奖励。另外, 每周一次, 用废弃的旧沙子和用异丙醇擦拭杯子来清洁陶瓷杯。</li>
		<li>准备下一次试验的砂杯, 根据随机脚本重新排列顺序, 并引诱正确的杯子。</li>
	</ol>
</li>
</ol>4. 挖掘砂杯和适应测试仪器的培训
<ol><li>倒入大约½杯的打砂成陶瓷杯 (直径6.99 厘米, 8.81 厘米高)。</li>
	<li>把满是沙子的杯子放在每个老鼠家笼子里过夜。 注:老鼠被要求成功地在杯子里挖, 并且连续两个晚上吃奖励过夜。</li>
	<li>习惯鼠在仪器中放置5分钟以检测仪器。<ol>
<li>在整个仪器中传播食物奖励, 鼓励仪器的探索和通过敞开的门旅行。确保大鼠不连续两天在室内小便、大便或冷冻。</li>
	</ol>
</li>
	<li>转移无嗅到的沙子填充杯子与食物报偿到测试仪器。确保老鼠成功地在沙子中挖掘, 并至少在连续两天内获得奖励, 不结冰的行为, 大便, 或排尿。如有必要, 取下陶瓷杯, 将谷物放进室内。</li>
</ol>5. 匹配样本 (MTS) 培训
<ol><li>用¼茶匙的香料 (肉桂、咖啡、辣椒粉和百里香) 混合大约3杯沙子, 创建4种不同的香味砂桩。为了控制气味线索, 在不正确的选择杯中粉碎一条奖励。<ol>
<li>每星期再混合一次有香味的沙子, 并储存在玻璃容器中, 这样香味强度在测试日是一致的。</li>
	</ol>
</li>
	<li>将每½杯倒入4杯, 并在黑色接触纸 (MTS 任务区) 的半圆上排列杯子;图 1A)。确保所有的杯子都与门的距离相等。<ol>
<li>根据编程软件 "R" 上的输出, 在没有替换功能的情况下, 将杯子放在指定的位置。</li>
	</ol>
</li>
	<li>出于对主题的看法, 在正确的香味杯中埋葬一整片奖赏。埋葬½的奖励在一个相同的香味样品杯。埋葬所有的奖励至少一个½的方式下来的沙子的高度。<ol>
<li>伪随机选择正确的香味杯, 唯一的限制是, 没有人的气味是样品的香味在一排2倍以上。</li>
	</ol>
</li>
	<li>将主题放在样品室中, 用弯曲的刮刀 (90 度角的手柄托盘) 将试样杯降低。允许在样品杯中挖掘, 并检索½谷物奖励。当主题开始吃奖励的时候, 把杯子取出来。</li>
	<li>一旦主题已经结束了吃的奖励, 提起纸板室分流器, 并打开门的测试室作为主体进入延迟室。</li>
	<li>允许主题通过挖掘来 "选择" 其中一个杯子。一旦主题作出选择, 并获得奖励, 如果存在, 删除该主题从会议厅和地方在转移笼。老鼠可能嗅到每个杯子, 但他们只允许一个选择, 通过开掘表明。<ol>
<li>在4分钟内, 用70% 异丙醇清洗房间。真空的任何沙子, 可能已经溢出和补充砂杯, 如果需要的话。</li>
		<li>获得下一个样品杯和诱饵的样品和测试杯。如果主题选择不正确的杯子在早先试验, 去除和放弃未吃的报偿。</li>
	</ol>
</li>
	<li>确保受试者每天接受一个6试验的 MTS 会话。为了通过 MTS 训练, 受试者必须在连续两个疗程中选择正确的三杯六次试验。 注:这可以确保性能高于机会, 但低于4选择测试的上限。</li>
</ol>6. 延迟滴定法在延迟匹配样品 (DMTS)
<ol><li>继续与以前的 MTS 进行培训 (从0秒的延迟开始)。 注:性能必须保持在40和70% 之间正确, 以确保性能远远高于机会 (25%), 但低于上限。在这个范围内的性能允许受试者在记忆相同的情况下, 当他们忘记样本时, 会遇到实例。这创造了一个机会, 利用下降测试的反应, 有利于这个主题, 当选择试验被引入。</li>
	<li>如果连续 2 6 次试验期的性能超过 70%, 则将延迟时间从0增加到三十年代。<ol>
<li>在延迟不是0秒的试验中, 用秒表计时每次延迟。在主体消耗完样品后, 提起纸板隔板墙, 允许进入延迟室。关闭纸板隔板, 开始计时秒表的延迟时间。</li>
		<li>三十年代后, 打开测试室的门, 门一旦进入测试室就关上门。</li>
		<li>像以前一样进行审判的其余部分。</li>
	</ol>
</li>
	<li>如果连续 2 6 次试验会话的性能继续保持在70% 以上, 则将随后的延迟增加1.5 倍于上一次延迟。</li>
	<li>如果连续2个会话的性能低于 40%, 则延迟将返回到以前使用的时间。</li>
	<li>如果连续2个会话的性能保持在40% 和70% 之间, 则将该延迟设置为特定于主题的标准延迟。</li>
</ol>7. 谢绝使用培训
<ol><li>每届 DMTS 训练后, 再放一杯不同质地和颜色的沙子, 在香味的沙杯对面。用不太喜欢的奖励, ¼的谷物, 这将是下降杯诱饵。从房间里取下香味的沙子杯。</li>
	<li>在 DMTS 训练期间进行试验, 允许受试者取样随机的香味杯, 并在延时室等待特定的延迟。</li>
	<li>允许在无味的沙杯中挖掘并取回奖励。然后从会议厅中删除该主题。这将被认为是 "被迫下降" 的审判。</li>
	<li>在每次 DMTS 训练后进行两次强制下降试验。</li>
</ol>8. 测试: 强迫vs选择试验
<ol><li>使用标准延迟测试 DMTS 的对象。每天对每一个12审判的课程进行测试。<ol>
<li>随机选择8的试验是 "选择" 的试验, 并把下降杯对面的香味杯。使用随机化脚本来确定试验类型的顺序, 约束条件是不超过3个选择或强制试验连续发生。 注:余下的4项试验将被 "强迫", 而下降的杯子从会议厅中移除。</li>
		<li>根据受试者的标准延迟, 可以在一只老鼠的过程中测试2或3个主语。在1主题的过程中, 运行第二个主题的审判。每次试验后确保用异丙醇清洗室。</li>
		<li>测试主题十次。</li>
	</ol>
</li>
	<li>在每次试验中, 一旦老鼠进入延迟室, 确保他要么向左转, 要么拒绝测试, 要么向右转以进行测试 (见图 1A; 自上而下的照片)。记录如果老鼠从房间的一侧移动到另一个 (例如"决定接受测试", 然后再遍历到房间的下降测试一侧)。 注:因此, 在遇到四种选择气味之前, 老鼠会做出一个前瞻性的青少年元记忆选择, 因为他选择接受或拒绝测试 (见结果纸19 , 以便彻底讨论并发与前瞻性青少年元记忆测试)。</li>
	<li>在两次测试和监测下降试验使用后连续计算强迫试验的准确性。<ol>
<li>如果在两个疗程的过程中准确度提高了70% 以上, 则停止测试并返回延迟滴定, 延迟时间更长。</li>
		<li>如果拒绝测试使用的方法始终是0% 或 100% (超过2个会话), 则暂停测试并考虑增加或降低下降测试响应的值, 以确保下降率介于10% 和40% 之间。</li>
		<li>如果2次会议的准确度低于 40%, 则停止测试并返回延迟滴定, 延迟时间缩短。</li>
		<li>在40% 和70% 之间, 对于连续两次的实验, 在强制试验的准确性上仍然保持正确。</li>
		<li>在整个测试过程中, 为了确保老鼠的喜好保持不变, 每周执行偏好检查 (2 次试验)。上面描述了首选项检查。</li>
	</ol>
</li>
</ol>9. 建议的综合测试
<ol><li>泛化测试 (如结果纸19中所描述的) 可以用来确定元认知响应是否在没有与外部刺激的单个关联可能控制响应的条件下传输。<ol>
<li>在无样本探针试验中试验大鼠, 未出现样本阶段, 但试验的 RI 和试验阶段与正常试验相同。随机混合4探针试验, 8 例正常样本试验, 构成12试验期。测试主题10次。将无抽样试验的下降率与正常试验的下降率进行比较。</li>
		<li>对大鼠进行双样本试验, 在该实验中, 实验对象与原样品香味两次呈现, 以增加记忆强度。如上文所述, 随机混合探针试验与正常样品试验。比较双样本试验的下降率和正常试验的下降率。</li>
		<li>用三种不同的 RIs 测试大鼠: 标准延迟, 一个延迟 (如0 秒) 比标准延迟, 一个更长 (如4 分钟)。在一届会议的每一次延误中, 有四项选择试验和两次强迫试验, 这些会议由18项试验构成。在十疗程中随机混合所有试验和实验鼠。比较试验的下降率在三延迟, 其中记忆强度应该减少和增加, 分别。</li>
	</ol>
</li>
</ol>10. 数据分析
<ol><li>测试完成后, 为每个科目的强制和选择的试验计算平均比例正确。<ol>
<li>为了找出正确的强迫试验的比例, 将正确的强迫试验数量除以强迫试验总数。</li>
		<li>为了找出正确选择的试验的比例, 首先从选择试验的总数中减去试验的次数。将正确的选择试验的数量除以选择试验的总数减去每只老鼠的拒绝试验。</li>
		<li>对组数据进行 t 检验, 将正确的强制试验比例与正确选择的试验比例进行比较 (见图 2)。</li>
		<li>如果运行泛化测试, 则对正常试验与探针试验中的下降率进行 t 检验, 如9所述。对于混合延迟试验, 如9.1.3 中所述, 将强迫和选择的测试与反复测量方差分析与延迟和试验类型作为一个内标因子进行比较。此外, 对下降使用的单向方差分析, 以确定是否准确性随 RI 而变化。</li>
	</ol>
</li>
</ol>

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没有要报告的披露。

本文提出了一种用气味为基础的 DMTS 任务来评估大鼠元认知应答的新方法。由于嗅觉作为啮齿类动物的主要感觉, 使用气味是更好的视觉线索在老鼠和老鼠20,21。在沙地上挖掘是大鼠自然觅食的行为, 使其与啮齿动物的生态相关。
该议定书的一个关键步骤是确保有一个粮食奖励等级, 其中一个显然比另一种更可取。在研究的早期阶段, 老鼠被要求在假定为首选奖赏, 一块甜麦片和小标准食品颗粒之间进行选择。然而, 偏好测试产生了不分青红皂白的选择行为, 导致了数量的使用作为奖励变异的来源。大鼠选择了一整片谷物和一¼的谷物, 并可靠地青睐较大的数量。如果没有偏好, 无论是食品质量还是数量, 都没有动力去测试它的下降, 所以这一步是至关重要的。
该协议的另一个重要方面是建立适当的 RIs, 使 MTS 精度保持在40到70% 的范围内。如果延迟太短, 并且干扰是极小的-由于长的, 老鼠从未体验 "忘记" 样品。反之, 如果 RIs 太长, 并且太短, 测试变得太困难, 老鼠永远不记得样本。这种反应不仅可以通过对强迫试验的准确性来检测, 而且还可通过过低或过高的下降率来探测。受试者应减少10-50% 的选择试验: 如果一个主题下降每一个试验或从来没有下降, 数据不能解释为积极或消极的存在青少年元记忆和下降测试反应的相对价值需要重新评估.在整个实验中, 强迫试验的平均性能在两天的测试结束时一直被计算出来。当老鼠通过更多的 DMTS 试验取得进展时, 它们的性能可能会提高, 这将启动一个返回延迟滴定 (见图 1B), 这样可以适当地增加延迟, 从而使精确度保持在70% 以下正确。如果样本的内存一直保持在70% 以上, 请考虑减少。如果样品的记忆保持在40% 以下, 考虑增加和/或增加气味样品大小从四到10-20 气味减少记忆干扰。
根据使用的大鼠的应变, 住房环境和光黑暗的循环动物被安置在, 记忆可能是微弱或更强的。与其他实验室大鼠菌株, 如 Dawleys 或斯科斯, 长埃文斯被称为获得认知任务, 相对较少的试验22,23。因此, 它可能会采取更多的适应阶段, 以及更多的培训, 其他啮齿动物学习的任务。目前研究中的大鼠被安置在高度丰富的环境中, 有锻炼的机会, 所以他们的记忆可能相对强24,25,26。这里使用的老鼠也被安置在一个反向的光黑暗周期, 这可能会增加认知性能, 因为他们在他们的黑暗周期测试。
自适应使用的下降测试反应, 表明在选择的性能显著高于强制测试表明元认知反应。然而, 元认知反应可能是依赖于内部记忆线索或外部线索8。为了确定是否实际上自适应地使用下降测试响应是内部记忆线索的结果, 实验者应该设法消除尽可能多的外部线索, 尽可能8,14。外部线索被认为是任何公共可用的测试特定的刺激或线索, 受试者可以用来指导下降测试使用8。此任务中潜在的外部提示包括单个气味的相对显著性、样本持续时间、响应延迟和 RIs 的持续时间。概括测试如这里所述, 更详细地说, 结果纸19有助于排除基于外部线索 (如环境提示关联) 的行为响应, 因为实验中任务参数的提示不一致。
在这个范式中, 老鼠需要作出决定, 以采取或拒绝测试之前的记忆测试提出 (见 8.2), 基本上消除了潜在的反应竞争控制使用的下降测试反应。响应竞争是在元认知选择与主记忆测试8同时出现的情况下, 根据正确测试选项的视觉或气味进行测试的倾向。要求老鼠在遇到记忆测试之前选择接受或拒绝测试本身会显著降低响应竞争控制使用下降测试响应的可能性。然而, 这种设计的一个改进是让老鼠选择采取或拒绝测试, 在一个单独的房间与记忆测试本身的一个特定的反应。这将进一步增加这种范式的前瞻性, 这可能使消除外部线索8、19更加成功。未来研究的另一项改进是, 如果可能的话, 在老鼠进入测试室的时候记录反应滞后时间, 以使主语向左或向右转 (分别下降或进行测试)。这将使研究人员能够确定响应延迟是否作为一个外部提示对象而被学会与特定的响应超时 (见8) 相关联。如果和当受试者改变他们的反应 (见 8.2; 例如从该房间的 MTS 选择一侧到下降测试区或反之亦然), 也会有启发性。正如结果文件中所讨论的, 我们只观察了一次这种行为, 这就增加了一个结论, 即大鼠一旦进入测试室, 就决定采取或拒绝测试, 而在他们遇到记忆测试本身之前, 他们做了系统取样每种选择气味。
因为在这个范式中测试的老鼠转移了元认知反应跨多个泛化测试, 它的结论是, 老鼠有能力青少年元记忆-他们可以监测他们的记忆样本19。对于其他研究小组来说, 测试这种范式或类似的适应性是很重要的, 因为需要从不同实验室得到确凿的证据, 才能断定老鼠有能力进行元认知应答, 并且行为由内部而不是外部提示控制。测试这一重要的记忆能力, 这与年龄下降, 可能证明是有用的翻译方法研究记忆功能障碍, 如转基因小鼠和阿尔茨海默病模型。这将是特别卓有成效的研究人员使用这种行为范式结合脑损害, 暂时性脑灭活或活化 (如光遗传学, chemogenetics), 并与电生理记录, 作为这些调查可以阐明元认知过程的神经机制。

青少年元记忆是评估2,3,4的记忆强度的能力。虽然人类一直表现出这种认知能力, 但在非人类动物中存在青少年元记忆的争论,例如5,6。测试青少年元记忆可能性的一个常用方法是用一个下降测试选项来呈现非人类动物, 如果选择的结果是有保证的, 但报酬比对测试的正确反应的奖励要小。如果受试者能够监控他们的记忆状态, 那么他们应该在记忆薄弱时拒绝测试 (查看:7,8,9,10)。
在过去的几十年中, 为人类和非人灵长类11和鸽子12开发的程序已经适应了几种方法, 以进一步测试在猴子中存在的青少年元记忆1,2,13.也许最重要的研究青少年元记忆的猴子是汉普顿的1延迟配对样本测试 (DMTS)。在 MTS 任务的训练之后, 样品和测试之间的延迟滴定, 使困难水平是中间的, 因此猴子有机会记住和忘记样品。延迟后, 猴子被提出了拒绝测试的选项。结果表明, 受试者比强迫试验更准确地进行了试验, 提示猴子监测其1样本的记忆强度。
迄今为止, 大多数青少年元记忆的实验范式都利用视觉或听觉刺激, 这并不奇怪, 因为最受考验的物种是高度视觉的非人类灵长类动物,例如11、14。在以前对大鼠元认知的有限研究中, 使用的感官模式也有视觉或听觉15,16,17,18。鉴于最近在使用基于气味的任务时对元认知反应的有力证据,19, 可能是由于缺乏标准化的范例而导致老鼠先前模棱两可的结果。我们设计了一个范式19 , 它的主要意义, 嗅觉, 通过适应 DMTS 范式类似的猴子在1以上描述。这里描述的任务包括在沙子填充的杯子里挖, 以获得食物奖励, 利用啮齿动物的自然挖掘行为。
青少年元记忆可以评估在大鼠使用气味为基础的 DMTS 任务在这里提出。首先, 我们建立了一个食物奖励的等级体系, 因此受试者表现出对更有价值的奖赏的明确偏好。然后, 老鼠被训练去挖一杯香沙来获得奖赏。在展示香味的样品杯后, 受试者必须选择四个明显的香味杯之一。奖励被埋葬在杯子的底部与样品相同的气味。在大鼠精通基本 MTS 测试后, 保留间隔 (RIs) 为每只老鼠滴定, 使性能在40% 和70% 成功率之间。在2/3 的试验中, 老鼠被给予选择, 以减少记忆测试, 为一个不太喜欢的奖励。对大鼠的元认知能力进行了比较, 评估了试验中的实验结果 (选择试验) 对大鼠被迫接受试验的试验 (强迫试验)。
虽然只有长埃文斯大鼠在这种模式下测试, 这项任务可以用来测试其他菌株的实验室大鼠和小鼠进一步研究青少年元记忆的啮齿动物物种。这也将有利于测试老龄啮齿动物或转基因小鼠的阿尔茨海默氏病的病理, 以更好地了解记忆障碍如何影响元认知过程。

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			<td height="42" style="height:42px;width:216px;">Colored Sand&#160;</td>
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testing for metacognitive responding using an odor-based delayed match-to-sample test in rats

青少年元记忆包括认知能力来评估自己记忆的力量。为了探讨青少年元记忆在非人类动物中的可能性, 已经创建了许多行为任务, 其中许多都利用一个选项来拒绝记忆测试。为了评估大鼠的青少年元记忆, 我们利用了这一下降试验选项范式, 将以前的视觉延迟匹配测试 (DMTS)1,2开发为灵长类动物的气味为基础的测试, 适合啮齿目动物。首先, 通过在一杯有香味的沙子中挖掘, 给老鼠一个样品来记住。延迟后, 大鼠有四个明显的香味杯, 其中一个包含相同的气味在样品中体验;如果这个匹配的杯子被选择, 则老鼠获得一个优选, 更大的报偿。选择任何其他三不匹配的充满沙子的香水杯, 结果没有奖励。保留间隔是单独滴定的, 这样, 主题执行40和70% 之间的正确, 因此确保老鼠有时记得, 有时忘记的样本。在这里, 青少年元记忆的操作定义是通过行为响应区分内存存在和缺失的能力。为此, 在2/3 的试验中, 除了四选择杯 (选择试验) 外, 还提出了减少的选择。如果下降测试选项-一个无味的彩色砂杯, 被选中, 主题收到一个较小的首选奖励, 并避免记忆测试。在余下的第三次试验中, 下降试验选项是不可用的 (强迫试验), 导致被试者猜测正确的杯子时, 样本被遗忘。在选择测试中, 知道何时记住的科目应该在内存不足时选择 "谢绝" 选项, 而不是进行测试并选择错误。因此, 与强制记忆测试相比, 所选测试的性能显著提高, 这表明了对衰落测试响应和元认知响应的自适应使用。

为了研究该学科的元认知应答, 正确的强迫试验 (基线记忆表现) 的比例与正确选择的试验的比例, 或有下降选择的试验相比较。与强迫试验相比, 所选试验的准确度明显较高 (图 2A) 显示阳性结果。这些结果表明, 大鼠可能是监测他们的记忆状态和下降的任务时, 他们的记忆是薄弱的, 导致提高准确性时, 选择了下降的选择是在选定的试验。
如果与强迫试验相比, 在选择的试验中性能没有显著提高, 结果是无效的。这种假设缺乏证据的青少年元记忆在大鼠 (图 2B) 表明, 如果没有使用下降测试选项时, 它本来可以适应这样做: 在试验中, 该样本被遗忘。
如果进行了泛化测试, 如果受试者减少无样本试验, 显著地减少了样本试验, 并显著降低了双样本试验, 则会显示元认知性能。拒绝使用应显示与内存准确性的反向关系, 如对强迫试验的准确性所示。如果强迫测试的准确性不随试验条件而变化, 则更改参数 (如增加 RI), 因为无法确定对象是否 metacognitively 或不行为。在混合延迟测试中, 对所选测试的准确性应显著高于强制测试的准确性, 而受试者更容易降低测试时间, 并且最不可能在较短的 ris 下降低测试。有关泛化测试和相应解释19的详细信息, 请参阅结果文件。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/57489/57489fig1v2.jpg"> 图 1.实验仪器和工艺的照片。A)从19改编的器具的照片, 允许使用。B)实验过程。第一步的习惯, 教老鼠挖砂灌杯, 发生在两个晚上。接下来, 大鼠习惯于实验仪器, 并被训练在那里挖砂杯。然后, 受试者可以选择两个奖励, 代表一个绿色的三角形和红新月 (整个谷物与一季度谷物的一块)。在老鼠表现出一个奖励的显著偏好后, 他们过渡到 MTS 训练, 其中杯子充满了香味的沙子, 代表了肉桂, 辣椒粉, 百里香和咖啡的图像。如果两届会议的成功率低于 50%, 则主题继续培训。如果大鼠的成功率为50% 或以上, 则研究对象会延迟滴定。在延迟滴定过程中, 大鼠继续进行 MTS 训练, 但样品和试验之间的延迟 (从0秒开始)。如果大鼠的记忆精度高于 70%, 则 RI 增加。如果准确度低于 40%, RI 减少, MTS 培训继续进行。这个循环一直持续到老鼠的精确度在40% 和70% 之间连续两个会话。在这一点上, 老鼠的实验测试, 包括随机混合强迫和选择试验。<a href="/files/ftp_upload/57489/57489fig1v2large.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/57489/57489fig2.jpg"> 图 2.积极的结果表明元认知反应和假设无效结果表明缺乏元认知反应。A)积极的结果从19适应了与允许。强迫和选择试验的平均比例正确来自9只大鼠在120次试验中的表现。* = p < 05 配对双尾 t 检验。与强迫试验相比, 在选择的试验中正确回答的试验比例明显较高, 表明了对下降试验反应的适当使用。B)在强迫和选择的试验中正确回答的同等比例的试验表明, 在记忆强度的基础上没有下降的试验。机会, 0.25, 由虚线指示。误差线表示方法的标准错误。<a href="/files/ftp_upload/57489/57489fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Testing for Metacognitive Responding Using an Odor-based Delayed Match-to-Sample Test in Rats at JoVE.com

Testing for Metacognitive Responding Using an Odor-based Delayed Match-to-Sample Test in Rats

该协议描述了一种调查啮齿类动物青少年元记忆或记忆感知的可能性的方法。基于气味的延迟匹配样本范式是一种新的、与生态相关的行为测试, 用于确定啮齿类动物在认知监测其记忆状态强度的基础上自适应地反应的程度。

用基于气味的延迟配对试验对大鼠元认知应答的测试

Metamemory involves the cognitive ability to assess the strength of one&#39;s memories. To explore the possibility of metamemory in non-human animals, ...

testing-for-metacognitive-responding-using-an-odor-based-delayed

Nanyang Technological University

Research

JoVE Journal

Behavior

7.1K 次观看.  Providence College. 该协议描述了一种调查啮齿类动物青少年元记忆或记忆感知的可能性的方法。基于气味的延迟匹配样本范式是一种新的、与生态相关的行为测试, 用于确定啮齿类动物在认知监测其记忆状态强度的基础上自适应地反应的程度。

视频: Testing for Metacognitive Responding Using an Odor-based Delayed Match-to-Sample Test in Rats

The Successive Alleys Test of Anxiety in Mice and Rats

The plus-maze was derived from the early work of Montgomery. He observed that rats tended to avoid the open arms of a maze, preferring the enclosed ones. Handley, Mithani and File et al. performed the first studies on the plus-maze design we use today, and in 1987 Lister published a design for use with mice. 
Time spent on, and entries into, the open arms are an index of anxiety; the lower these indices, the more anxious the mouse is. Alternatively, a mouse that spends most of its time in the closed arms is classed as anxious. 
One of the problems of the plus-maze is that, while time spent on, and entries into, the open arms is a fairly unambiguous measure of anxiety, time in the central area is more difficult to interpret, although time spent here has been classified as &#x201C;decision making&#x201D;. In many tests central area time is a considerable part of the total test time.
Shepherd et al. produced an ingenious design to eliminate the central area, which they called the &#x201C;zero maze&#x201D;. However, although used by several groups, it has never been as widely adopted as the plus-maze. 
In the present article I describe a modification of the plus-maze design that not only eliminates the central area but also incorporates elements from other anxiety tests, such as the light-dark box and emergence tests. It is a linear series of four alleys, each having increasing anxiogenic properties. It has given similar results to the plus-maze in general. Although it may not be more sensitive than the plus-maze (more data is needed before a firm conclusion can be reached on this point), it provides a useful confirmation of plus-maze results which would be useful when, for example, only a single example of a mutant mouse was available, as, for example, in ENU-based mutagenesis programs.

The plus-maze measures anxiety-like behaviour in rodents. There are two opposite closed and two opposite open arms; anxious rodents avoid the open arms. The central area is neither completely open nor closed, so time spent here is ambiguous and difficult to interpret. Here a modification of the plus-maze protocol eliminating this area is described.

在小鼠和大鼠连续的小巷测试焦虑

The plus-maze was derived from the early work of Montgomery. He observed that rats tended to avoid the open arms of a maze, preferring the enclosed ones.  ...

科研

行为学

A Procedure to Study the Effect of Prolonged Food Restriction on Heroin Seeking in Abstinent Rats

In human drug addicts, exposure to drug-associated cues or environments that were previously associated with drug taking can trigger relapse during abstinence. Moreover, various environmental challenges can exacerbate this effect, as well as increase ongoing drug intake.
The procedure we describe here highlights the impact of a common environmental challenge, food restriction, on drug craving that is expressed as an augmentation of drug seeking in abstinent rats.
Rats are implanted with chronic intravenous i.v. catheters, and then trained to press a lever for i.v. heroin over a period of 10-12 days. Following the heroin self-administration phase the rats are removed from the operant conditioning chambers and housed in the animal care facility for a period of at least 14 days. While one group is maintained under unrestricted access to food (sated group), a second group (FDR group) is exposed to a mild food restriction regimen that results in their body weights maintained at 90% of their nonrestricted body weight. On day 14 of food restriction the rats are transferred back to the drug-training environment, and a drug-seeking test is run under extinction conditions (i.e.&#160;lever presses do not result in heroin delivery).
The procedure presented here results in a highly robust augmentation of heroin seeking on test day in the food restricted rats. In addition, compared to the acute food deprivation manipulations we have used before, the current procedure is a more clinically relevant model for the impact of caloric restriction on drug seeking. Moreover, it might be closer to the human condition as the rats are not required to go through an extinction-training phase before the drug-seeking test, which is an integral component of the popular reinstatement procedure.

A procedure that allows a demonstration of robust augmentation of drug seeking in food-restricted rats is described. Following heroin self-administration training, rats go through an abstinence period, in a drug-free environment, during which they are mildly food restricted. Drug seeking is then tested in the drug-associated environment.

一个过程来研究延长食品的限制对海洛因寻求戒断大鼠的影响

In human drug addicts, exposure to drug-associated cues or environments that were previously associated with drug taking can trigger relapse during ...

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 ...

A Procedure to Observe Context-induced Renewal of Pavlovian-conditioned Alcohol-seeking Behavior in Rats

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate drug-seeking behavior and prompt relapse in abstinent drug users. We have developed a procedure to study the behavioral and neural processes that mediate the impact of context on alcohol-seeking behavior in rats. Following acclimation to the taste and pharmacological effects of 15% ethanol in the home cage, male Long-Evans rats receive Pavlovian discrimination training (PDT) in conditioning chambers. In each daily (Mon-Fri) PDT session, 16 trials each of two different 10 sec auditory conditioned stimuli occur. During one stimulus, the CS+, 0.2 ml of 15% ethanol is delivered into a fluid port for oral consumption. The second stimulus, the CS-, is not paired with ethanol. Across sessions, entries into the fluid port during the CS+ increase, whereas entries during the CS- stabilize at a lower level, indicating that a predictive association between the CS+ and ethanol is acquired. During PDT each chamber is equipped with a specific configuration of visual, olfactory and tactile contextual stimuli. Following PDT, extinction training is conducted in the same chamber that is now equipped with a different configuration of contextual stimuli. The CS+ and CS- are presented as before, but ethanol is withheld, which causes a gradual decline in port entries during the CS+. At test, rats are placed back into the PDT context and presented with the CS+ and CS- as before, but without ethanol. This manipulation triggers a robust and selective increase in the number of port entries made during the alcohol predictive CS+, with no change in responding during the CS-. This effect, referred to as context-induced renewal, illustrates the powerful capacity of contexts associated with alcohol consumption to stimulate alcohol-seeking behavior in response to Pavlovian alcohol cues.

A procedure to study the capacity of an alcohol associated environmental context to trigger the renewal of alcohol-seeking behavior in rats is described.

一个过程来观察大鼠巴甫洛夫空调酒精寻求行为的背景下引发的重建

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate ...

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 ...

Testing for Odor Discrimination and Habituation in Mice

This video demonstrates a technique to establish the presence of a normally functioning olfactory system in a mouse. The test helps determine whether the mouse can discriminate between non-social odors and social odors, whether the mouse habituates to a repeatedly presented odor, and whether the mouse demonstrates dishabituation when presented with a novel odor. Since many social behavior tests measure the experimental animal&#8217;s response to a familiar or novel mouse, false positives can be avoided by establishing that the animals can detect and discriminate between social odors. There are similar considerations in learning tests such as fear conditioning that use odor to create a novel environment or olfactory cues as an associative stimulus. Deficits in the olfactory system would impair the ability to distinguish between contexts and to form an association with an olfactory cue during fear conditioning. 
In the odor habitation/dishabituation test, the mouse is repeatedly presented with several odors. Each odor is presented three times for two minutes. The investigator records the sniffing time directed towards the odor as the measurement of olfactory responsiveness. A typical mouse shows a decrease in response to the odor over repeated presentations (habituation). The experimenter then presents a novel odor that elicits increased sniffing towards the new odor (dishabituation). After repeated presentation of the novel odor the animal again shows habituation. This protocol involves the presentation of water, two or more non-social odors, and two social odors. In addition to reducing experimental confounds, this test can provide information on the function of the olfactory systems of new knockout, knock-in, and conditional knockout mouse lines.

This manuscript describes a protocol to examine the olfactory system of rodents. The olfactory habituation/dishabituation test will allow investigators to determine whether a mouse habituates to a repeatedly presented odor and whether the mouse demonstrates dishabituation when presented a novel odor.

在小鼠试验的气味辨别和习惯化

This video demonstrates a technique to establish the presence of a normally functioning olfactory system in a mouse. The test helps determine whether the ...

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, ...

The Social Dimension of Stress: Experimental Manipulations of Social Support and Social Identity in the Trier Social Stress Test

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations supportive behavior of other people as well as positive social relationships can act as powerful resources to cope with stress. In order to study the interplay between these variables, this protocol describes two effective experimental manipulations of social relationships and supportive behavior in the laboratory. In the present article, these two manipulations are implemented in the Trier Social Stress Test (TSST)&#8212;a standard stress induction paradigm in which participants are subjected to a simulated job interview. More precisely, we propose (a) a manipulation of the relationship between different protagonists in the TSST by making a shared social identity salient and (b) a manipulation of the behavior of the TSST-selection committee, which acts either supportively or unsupportively. These two experimental manipulations are designed in a modular fashion and can be applied independently of each other but can also be combined. Moreover, these two manipulations can also be integrated into other stress protocols and into other standardized social interactions such as trust games, negotiation tasks, or other group tasks.

Previous research on the social dimension of stress has focused on two important variables: social identity and social support. This protocol introduces an effective experimental manipulation of these two social variables and describes their implementation in a standard stress induction paradigm (Trier Social Stress Test).

应力的社会问题:社会支持和社会认同的特里尔社会的实验操作压力测试

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations ...

The Rodent Psychomotor Vigilance Test (rPVT): A Method for Assessing Neurobehavioral Performance in Rats and Mice

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article describes a rodent version of the PVT&#8212;termed the &#34;rPVT&#34;&#8212;that measures similar aspects of attention (i.e., performance accuracy, motor speed, premature responding, and lapses in attention). Data are presented that demonstrate both the short- and long-term usefulness of the rPVT when employed with laboratory rats. Rats easily learn the rPVT, and learning to perform the basic procedure takes less than two weeks of training. Once acquired, rat performances in the rPVT show a high degree of similarity to these same performance measures in the human PVT, including similarities in, lapses in attention, reaction times, vigilance decrements across session time (i.e., the human &#34;time-on-task&#34; effects), and the response-stimulus interval (RSI) effect described for humans. Thus the rPVT can be an extremely valuable tool for assessing the effects of a wide range of variables on sustained attention quite similar to human PVT performances, and thus can be useful for developing novel treatments for neurobehavioral dysfunctions.

The Rodent Psychomotor Vigilance Test rPVT: A Method for Assessing Neurobehavioral Performance in Rats and Mice

A rat version of the human Psychomotor Vigilance Test (PVT) is described that measures aspects of attention similar to those measured with the human PVT, including aspects of human vigilance such as performance accuracy, motor speed, premature responding, and lapses in attention.

鼠类精神运动警觉测试（rPVT）:一个在大鼠和小鼠的神经行为评估性能的方法

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article ...

Testing Animal Anxiety in Rats: Effects of Open Arm Ledges and Closed Arm Wall Transparency in Elevated Plus Maze Test

The elevated plus maze test is a behavioral test for assessing animal anxiety in rodents. Although this test is widely applied in the field of behavioral science, conflicting outcomes are often provided from different laboratories. To identify reasons for the different outcomes, we previously focused on arm features, which differ between laboratories, most notably the presence/absence of ledges at the sides of open arms and the transparency/opaqueness of closed arm walls. In a previous report, we used a custom designed container to compare rat behavior on different combinations of open and closed arm designs under otherwise identical experimental conditions, and showed that differences in arm features interfere with experimental outcomes. In brief, open arm ledges significantly increased anxiety-like behavior in rats, while transparent arms may also have decreased this behavior. Furthermore, we verified a higher detection sensitivity of the effect of an anxiolytic drug in a combination of no-ledged open arm + opaque walled closed arm compared with a combination of ledged open arm + transparent walled closed arm. In this report, we introduce our protocol for the elevated plus maze test, together with discussion of the key results from the previous report and our experimental experience. We believe this report will provide useful information for researchers who have employed or who plan to use the elevated plus maze in their studies.

Rats' anxiety-like behavior tested by the elevated plus maze varies highly depending on the testing environment. Here we show how apparatus settings, especially arm structure, affect behavior on the maze, and we explain a reliable protocol to detect effects of anxiolytic drugs. This report will contribute to anxiety research.

大鼠动物焦虑症的实验研究: 开放臂壁和闭合臂墙透明度对高加迷宫试验的影响

The elevated plus maze test is a behavioral test for assessing animal anxiety in rodents. Although this test is widely applied in the field of behavioral ...