这项工作全部由英国医学研究理事会授予的资助 (授予参考: MR/M007006/1) 授予 Sunjeev Kamboj 和拉维 Das。改进的记忆检索程序和线索图像的来源是由格雷斯大风。我们要感谢凯蒂. 沃尔什和何超仪 Hanniford 对试验程序的贡献。

所有程序均由伦敦大学学院研究伦理委员会批准, 并按照《赫尔辛基宣言》进行。在使用此方法之前获得所有必要的本地批准。
1. 味觉厌恶性无条件刺激的制备
<ol><li>使用微1毫升吸管, 添加80µL 铵苯甲酸溶液 (2.5% 在水中) 到一个混合玻璃。</li>
	<li>使用测量缸, 仔细测量120毫升的清洁水进入混合玻璃含有铵苯甲酸溶液。搅拌均匀。这产生120.08 毫升的0.067% 铵苯甲酸酯。</li>
	<li>均匀地把0.067% 铵苯甲酸溶液分成8干净, 完全相同的不透明杯。<ol>
<li>使用高精度电子秤将溶液分解成杯子。首先, 把一个空的杯子放到磅秤上, 把天平的重量调到零。慢慢地加入0.067% 铵苯甲酸溶液, 直到比例读 15 g. 重复这8杯。这将在每个杯子中提供等量的溶液。</li>
		<li>另外, 使用5毫升注射器, 添加15毫升的0.067% 铵苯甲酸酯溶液的塑料杯。这将在每个杯子中提供等量的溶液。</li>
	</ol>
</li>
	<li>在参加者到达之前, 在屏幕后面安排8杯苦涩饮料, 这样他们很容易被实验者接触到, 但是在参与者的视线之外。 注: 每个杯子的内容的消耗量担当味觉厌恶无条件的刺激 (UCS) 试验。</li>
</ol>2. 参加者的准备和饮水措施
<ol><li>在统计数据包中使用随机化命令 (例如, 使用 R47中的 "set. 种子" 和 "示例" 函数) 或适当的在线随机分配工具 (,如、random.org48, 将参与者随机化到其中一个以下干预组: (1) 检索 + pe (零售 + pe) (2) 检索 + 无 pe (不加网+ 无 pe) (3) 没有检索 + pe (不进行任何零售 + pe), 这样分配给每个组是相等的。</li>
	<li>确认危险饮酒与酒精使用障碍识别测试 (审核)49。本研究认为分数 > 10 是可以接受的。</li>
	<li>通过结构化临床访谈评估酒精依赖程度的诊断和统计 Manual-5 (无缺陷)50标准。考虑 < 4 项目编码为阈值水平可以接受 (即排除那些与当前酒精使用紊乱)。有关纳入和排除标准的更多信息, 请参阅9节。 注: 如果使用当前的刺激方案, 参与者必须首先自我报饮啤酒 (> 60% 总饮酒量)。确切的包含/排除标准将取决于研究的性质和目的。如果禁忌 counterconditioning 或接触酒精提示, 请确保参与者被排除在外。</li>
	<li>获得与会者的知情同意。</li>
	<li>使用酒精酒精测试装置, 以确保参与者不醉。如果读数超过0.001 毫克/升, 重新测试日或排除参与者。</li>
	<li>座椅的参与者在电脑前被用来显示刺激。</li>
	<li>评估状态 (在瞬间) 对酒精的渴求在基线和后再次接触的提示与酒精渴求调查表 (ACQ 现在)51和酒精消费量在过去一周的基线和后续 (天 8) 使用时间轴跟随返回酒精 (TLFB)52 。</li>
	<li>使用变化准备阶段和治疗渴望量表 (苏格拉底)53以确定饮酒的关注和准备变化, 厌恶倾向和灵敏度量表修订 (DPSS) 指数倾向和敏感性的厌恶54, 而负饮酒预期调查表 (NAEQ) 为负饮酒期望55。</li>
	<li>使用刺激演示软件来呈现所有的刺激, 在内存重新激活和 counterconditioning 过程 (步骤 3.2.2. 3.3.4 和 5.1. 到 6.4.)。</li>
</ol>3. 内存重新激活和控制非重新激活程序
注意: 请参见示意图的图 1 。
<ol><li>对于学员在零售 + pe 和零售没有 pe 组, 倒入150毫升的冷冻无酒精啤酒到一个品脱 (284 毫升) 玻璃, 并把这在表之间的参与者和显示屏。对于那些在没有零售 + PE 组, 倒入150毫升的冰镇橙汁到玻璃, 并把这个放在桌前的参与者。</li>
	<li>对于刺激评级, 目前的指示和刺激的相关检索/没有检索程序。 注意: 在发布的研究26中使用的确切说明可根据作者的要求提供。<ol>
<li>告诉与会者, 他们面前的玻璃杯中含有啤酒或橙汁, 适合他们的群体。通知他们, 他们将消耗这一饮料根据 on-screen 的指示后, 评级的一些图片的愉悦和影响 ' 冲动喝 ' 啤酒或果汁在他们面前。</li>
		<li>向参与者解释, 11 点的愉悦度从-5 (非常不愉快) 到 0 (既不愉快也不令人不快) 到 +5 (非常愉快) 运行。对于饮酒量的要求, 解释它从-5 (大大减少饮酒的冲动) 到 0 (对冲动没有影响) 到 +5 (大大增加饮酒的欲望)。指示参与者将所有评分大声。</li>
		<li>告诉参与者, 当他们喝啤酒 (或橙汁) 来指导他们的收视率时, 记得以前的例子。</li>
		<li>显示参与者的例子, 他们将看到当他们被要求消费的饮料在前面的 on-screen, 例如, "拿起饮料" 的黑色文本, "准备喝" 在蓝色的文本, "现在喝" 的绿色文本。指示参与者只在看到 "现在饮酒" 的时候才喝酒, 用绿色的文字写。</li>
		<li>以相关方式为正在使用的刺激演示软件部署重新激活/控制任务。计划的任务, 目前的条件刺激 (CS) 的图像为十年代每个 pseudo-randomized 的顺序。这项命令是由软件中的选项来决定的, 用于计划的任务, 并规定不超过两个连续的介绍啤酒 CS。只有当 "现在喝" 在绿色文本中时, 参与者才能饮用。</li>
		<li>记录参加者对四啤酒图像的愉悦/冲动评分 (酒精提示;cs + s) 和两个性奖励饮料的图像 (中性提示 CS s, 即咖啡和可乐) 的零售 + pe 和零售没有 pe 组, 或四橙汁的图像和两个 CS s 为无零售 + pe 组。</li>
	</ol>
</li>
	<li>
在体内评级和预测错误程序
	<ol>
<li>在所有的图像被评为, 直接参与者的注意, 在他们面前的饮料。要求他们对饮料本身的愉悦度 (看到) 进行评分, 以及它对他们目前的饮酒欲望有多大影响 (两者在以前使用的-5 到 +5 的比例)。<ol>
<li>让参与者想象一下, 他们会在-5 (非常不愉快) 到 5 (非常愉快) 的时候喝下饮料, 并对他们的感觉感到愉悦。</li>
			<li>指导参与者想象饮用饮料, 并将他们目前的消费欲望从-5 (至少我曾经想喝) 到 +5 (我最想喝的饮料)。注意不同的措辞到早先-5 到 +5 敦促标度。</li>
		</ol>
</li>
		<li>开始 on-screen 饮酒指示。<ol>
<li>对于所有的参与者, 分别显示前两个屏幕, 其中阅读 "拿起饮料" 和 "准备喝"。对于那些在零售 + pe 和没有零售 + pe 组, 显示最后的屏幕, 读 ' 停止!不要喝!参赛者不得饮用啤酒或橙汁, 产生阴性预测错误。</li>
			<li>对于那些在没有 PE 组, 显示最后的屏幕, 读 "现在喝", 并让参与者消费的啤酒。显示所有5秒的饮水指令屏幕。</li>
			<li>在饮用或不食用饮料后, 显示屏幕上的 "放下饮料并按下空格键继续"。确保遵守本指令。</li>
		</ol>
</li>
		<li>从参与者的视线中取出玻璃。</li>
		<li>让参与者大声地说出预期的/意想不到的前面的指令是从-5 (完全出乎意料) 到 5 (完全预期)。记下响应。</li>
	</ol>
</li>
</ol>4. 分心任务
<ol><li>将耳机放在参与者身上。</li>
	<li>按照下面列出的顺序立即执行一系列短期内存测试。<ol>
<li>首先, 播放 Rivermead 行为记忆测试56中的一个散文召回版本。通过要求参与者写下尽可能多的故事细节, 从而立即召回。</li>
		<li>按照此任务的标准协议管理数字范围任务57。</li>
		<li>管理语言和类别流利性任务58, 遵循任务完成的标准协议 (六十年代尽可能多地命名范例)。</li>
	</ol>
</li>
</ol>5. Counterconditioning 任务说明
<ol><li>在3步中概述的检索/无检索过程中使用的同一个刺激演示软件中实施 counterconditioning 任务, 以便它可以从检索/无检索过程中进行。 注意: 此任务的 pre-deployed 版本将在大多数操作系统上运行, 可根据作者的要求提供。</li>
	<li>开始 counterconditioning 任务的说明。<ol>
<li>通知参与者, 他们将查看一系列图片, 随后将显示在屏幕上的不同结果, 如另一张图片或 "现在喝" 字样。</li>
		<li>从屏幕后面的第一 UCS 饮料, 并把这个在参与者面前。通知参加者, 只要他们看到 "现在喝" 的字样, 他们就必须拿起杯子, 喝下里面所有的液体。告知参加者, 这些饮料可能味道很苦, 但并不有害。 注: 参加者必须不知道总的饮料或数量在整个 counterconditioning。</li>
		<li>指导参与者, 每当他们在每次试验中看到第一个 (CS) 图像时, 他们必须对-5 (非常令人不快) 到 +5 (非常愉快) 的图像的愉悦程度进行分析。</li>
		<li>让参与者使用键盘上的专用键来进行评分。使用键 [反斜线], [1], [2], [3], [4], [5], [6], [7], [8], [9] 并且 [0] 或交替的连续钥匙与贴纸安置在他们读 [-5], [-4], [-3], [-2], [-1], [0], [+ 1], [+ 2], [+ 3], [+ 4] 和 [+ 5], 分别。要提醒参与者进行此分级, 请在显示第一个图像 (CS) 时在屏幕上显示 "速率愉悦" 字样。</li>
		<li>告知参加者, 在结果发生后 (UCS 饮或另一张图片) 将结果的愉悦率在相同的-5 (非常令人不快) 到 +5 (非常愉快) 的比例。这提到苦涩饮料的消耗量 UCS 或表示法 UCS。 注意: 在进行这些分级之前, 任务中的下一个试验不会开始。</li>
		<li>重申, 参加者将因此对每项试验进行两次评分, 一个用于初始图像 (CS), 第二个用于结果 (UCS)。</li>
		<li>确保参与者了解这些说明并开始 counterconditioning 任务。</li>
	</ol>
</li>
</ol>6. 运行 Counterconditioning 任务
<ol><li>使用4啤酒刺激作为 cs + s (在检索任务中使用的4啤酒刺激) 和两个中性的刺激作为 cs s (咖啡和可乐图像中使用的检索/没有检索任务)。准备三 UCS "类型"。第一种类型: "味觉 UCS" 包括在屏幕上出现的 "现在喝" 的字样, 以及参与者在他们面前消耗的一杯苦涩的液体。第二种类型: "图案 UCS", 包括下面详述的一张令人作呕的图片。第三种类型: "中性 UCS" 由下面详述的神经图像组成。<ol>
<li>对两个啤酒 cs + s (指定 "饮料 cs + s") 与味觉 UCSs (这些在表 1的 "UCS 类型" 列中标记为 "味觉")。下面这些 CSs, 让 "现在喝" 的字样出现在屏幕上, 并要求参与者使用摆在他们面前的苦味液体杯。对二 (指定的 "图片 CS + s") 与视觉反感诱导 UCSs 从国际情感图片系统 (IAPS; 三图片: 9301.jpg, 9325.jpg, 9405.jpg, 和第四来源从互联网描述一个伤口的人的脚, 是出没与蛆;这些 CSs 在表 1中被标记为 "画报"。 注: 啤酒 CS 对结果的分配应是随机的, 并在开始收集数据之前进行。然后将 CS 分配给结果类型, 在所有参与者中保持不变。试验和试验序列在所有实验组中是相同的。中性 CS s 的结果 UCSs 是中性价 IAPS 图像 (1020.jpg 和 1021.jpg)。所有配对都在100% 加固时间表上。</li>
		<li>对每一个图片 CS + s 与每一个视觉厌恶 UCS 图像一次 (8 试验共计)。对每个味觉 CS + s 与词 ' 现在喝 ' 和消耗量的味觉厌恶 UCSs (苦涩饮料) 四次 (8 试验共计)。对每个中性的 CS s 与中性 UCS 图片四次 (8 试验总计)。因此, 总共执行24项试验。</li>
	</ol>
</li>
	<li>通过按下一个专用键 (空格键) 来开始 counterconditioning 任务, 一旦参与者了解到他们需要对每个试验进行两次评分: 一个用于初始图片 (CS), 一个用于结果 (UCS)。</li>
	<li>对所有参与者使用 pseudo-randomized 的试用订单。表1中给出了一个工作实例, 所有24项试验都是在一次会议上完成的。 注: 本命令确保在连续的试验中不会发生相同的 UCSs, 同时不超过两次相同 CS 的试验连续发生。但是, 只要在参与者之间保持一致, 就可以使用满足这些要求的其他试用排序。<ol>
<li>让参加者在无零售 + PE组率四啤酒 CS + s 为愉悦一次, 立即在开始之前 counterconditioning。 注意: 这将提供该组的基线评级, 并确保前到 CSs 的 counterconditioned 的卷在组之间是相同的。没有必要在 "无 pe " 或 "零售 + pe" 中执行此项, 因为它们已经对啤酒图像进行了分级。</li>
		<li>采取第一 UCS 饮料从后面的屏幕, 并放置在前面的参与者, 准备 "现在喝" 的指示。</li>
		<li>将表 1中的第一个 (CS) 图像显示为6秒。让参与者在每一次试验的第一个 (CS) 图像呈现时, 他们在6秒内从-5 (非常不愉快) 到 +5 (非常愉快) 的图像的愉悦程度。</li>
		<li>让参与者使用键盘上的专用键来进行评分。使用键 [反斜线], [1], [2], [3], [4], [5], [6], [7], [8], [9] 并且 [0] 或交替的连续钥匙与贴纸安置在他们读 [-5], [-4], [-3], [-2], [-1], [0], [+ 1], [+ 2], [+ 3], [+ 4] 和 [+ 5], 分别。要提醒参与者进行此分级, 请在显示第一个图像 (CS) 时在屏幕上显示 "速率愉悦" 字样。</li>
		<li>提出 6 s 的 "现在喝" 的指示, 让参与者拿起杯子并消耗液体。确保在每味觉 UCS counterconditioning 试验中消耗铵苯甲酸溶液的全部用量。一旦一杯饮料完成, 取出空杯, 取而代之的是下一个15毫升的铵苯甲酸酯溶液杯。</li>
		<li>让参加者对味觉 UCS 结果的愉悦率在相同的-5 (非常令人不快) 到 +5 (非常愉快) 的比例在他们消耗了整个15毫升后的 CS 率。 注: UCS 评级没有时间限制。一旦评分, 一个单一的试验是完整的。</li>
		<li>在序列中显示下一个 CS (一张中性图片, 在表 1中 CS-1), 用于6秒。让参与者在这段时间内使用-5 的响应键 (非常令人不快) 到 +5 (非常愉快), 如前所述, 使他们发现图像的愉悦程度。</li>
		<li>继续介绍余下的22项试验。 注意: 整个 counterconditioning 任务大约需要10分钟。</li>
	</ol>
</li>
	<li>完成后, 让参加者消费两个方块的牛奶巧克力, 以消除残留苦味的铵苯甲酸酯溶液。指示他们允许巧克力融化在他们的舌头上。</li>
</ol>7. 一周的后续措施
<ol><li>在检索/counterconditioning 过程之后的一周内安排此会话。<ol>
<li>喝酒精读的酒如果读数超过 0.001, 重新测试日或排除参与者。记录是否获得阳性读数 (> 0.001 毫克/升)。</li>
	</ol>
</li>
	<li>目前所有的 CSs 使用的 counterconditioning 任务加三新的啤酒图片和三新的葡萄酒图片 (10 酒精图像总共) 为十年代每一个 pseudorandomized 的顺序。让参与者的愉悦率从-5 (非常令人不快) 到 +5 (非常愉快) 和影响的冲动喝啤酒从-5 (大大减少冲动), 以 + 5 (大大增加敦促) 为每一个形象。在纸上记录他们的反应。 注: 新的图片评级被用来评估在啤酒刺激类和其他类型的酒精影响的泛化。</li>
	<li>管理 ACQ-现在49, 酒精 TLFB52的前一周, DPSS-R54, 和苏格拉底53。 注: 同样的后续措施可以重复使用, 以进行额外的、延伸的跟进。</li>
</ol>8. 点探针任务 (可选)
<ol><li>如果可以使用眼睛跟踪设置, 请在这个阶段用 counterconditioning 任务中使用的图像完成注意力偏置点探针任务。 注意: 此任务的详细信息在 Das et al.中给出28 , 此处不给出。此任务可根据作者的要求提供。</li>
</ol>9. 纳入和排除标准
<ol><li>对危险饮酒者使用下列纳入标准: 危险饮用 (在酒精使用紊乱识别试验中定义为分数 > 10, 但 < 4 项编码为3的无缺陷), 消耗 > 3 单位为女性, > 4 单位为男性在每周至少4天 (8 克纯酒精/单位), 流利的英语, 正常或 corrected-to-normal 的颜色视觉。</li>
	<li>使用以下排除标准: 年龄 < 18 和 > 65, 过去或现在诊断的药物或酒精使用紊乱, 任何目前治疗的精神健康问题, 目前的主要身体健康问题, 目前怀孕, 或母乳喂养。 注意: 精确的包含和排除标准可能会因使用本程序的测试人口而异。</li>
</ol>10. 数据预处理和分析59
<ol><li>为了分析 cs 的额定值, 在 counterconditioning 中计算每种 cs 类型的四个世纪。对于每个参与者, 这是通过简单地采取每两个类型的连续试验的平均值来实现的。例如, 按照表 1中给出的顺序, 平均测试1和试用3的分级, 以获得啤酒 CS+1 的第一个纪元。 注: 这将产生4的 "试验" 级别的每一个刺激 counterconditioning 评级数据。这样做是为了消除由于杂散顺序和预期效应而产生的 trial-to-trial 变化。</li>
	<li>评估所有数据的正常性、方差和度的一致性 (对于重复的措施, k> 2 比较)。<ol>
<li>在方差的 one-way 分析中违反方差的均匀性, 使用韦尔奇的 F 检验。在度被违反的情况下, 根据ε56的级别使用黄齐 Feldt 或温室 Geisser 校正。Winsorize 任何离群点 > 3 法令远离样本平均值为该变量到一个分数3法令的平均值。运行分析和没有这个过程评估结果的敏感性对过程和报告任何分歧。</li>
	</ol>
</li>
	<li>评估 counterconditioning 在响应味觉 cs + s 和图案 cs 时是否有任何差异。如果不是, 则折叠两个 CS 类型进行 counterconditioning 分析。如果存在差异, 请将 CS 类型作为 counterconditioning 数据分析的额外级别。</li>
</ol>

所有的作者都没有知识, 金融, 或生物医学的利益冲突披露。

该协议在本文中描述了一个酒精 MRM 检索程序, 显着产生预测错误的酒精传递 (检索 + PE), 以最大限度的概率, 自然主义酒精 MRMs 将动摇。该检索过程考虑了最近的实验工作, 证明了内存失稳的预测误差的必要性, 并参与了再过程60。在两篇论文28,46中, 它已被证明能从随后的行为干预 (counterconditioning 和认知重估) 中产生最深刻的酒精 MRMs 指数变化, 目前正在进一步在药物模型中得到验证。由于这一效果似乎与学习历史无关, 因此, 该过程比其他内存重新激活技术 (不包含 PE) 有了显著的改进, 并且应用作评估post-destabilization 减少适应饮酒行为的干预措施。根据再领域的空发现, 这种技术的细化尤为重要 (见下文)。此外, 该方法具有相当的应用意义, 可与多种 post-retrieval 再学习方法相结合。例如, 我们已经展示了对语义记忆和渴求的影响, 然后再重新评估, 一种典型的自适应情绪调节技术, 通常用于认知行为治疗46。
该程序易于修改, 以适应参与者组的特征。例如, 在 counterconditioning 期间使用的 CS + s 的刺激可以改变为葡萄酒的图片, 如果参与者主要是葡萄酒偏好。另外, 吸烟相关的刺激可以使用, 如果该程序是在烟草吸烟者。
有效的 counterconditioning 依赖于强有力的厌恶暗示。为了确保在再期间有足够的再学习, 选择可能引起强烈厌恶反应的视觉 UCSs。虽然这些都是视觉暗示, 但它们应该与味觉厌恶 (例如, 与污染相关的图像有关; 食物变质的图像)。如果使用一种替代的味觉 UCS (在这里使用的浓度以外的苯甲酸铵), 我们推荐一种强烈激活苦味反应的化合物。
虽然我们使用预期违规, 但理论上可以使用其他方法生成 PE, 尽管它们在稳定编码的奖赏记忆中的功效仍有待证实。我们有试用的替代程序, 涉及意外的酒精贬值 (使用意想不到的苦味酒精), 虽然我们没有证明令人信服的内存修改使用此过程46。
参与者对群体的随机化应防止特征问卷测量中的基线差异。然而, 如果是这种情况, 这些差异应被模拟为对结果的混合效应分析中的随机效应。作者建议, 补充方法是在检索和 counterconditioning 日之前添加一个额外的基线日。这可以更好地评估存在的群体差异, 这些差异的可靠性, 并给研究人员的选择分层随机分组, 以防止这种差异。
为了生成有效的 PE, 在检索过程中, 参与者必须经历一个相关的令人惊讶的事件。如此处所述, 如果使用预期违反行为, 请确保在这一步骤中观察参与者, 以免他们在被指示不小心饮用酒精的点。与此相关的是, 参与者不知道实验者在产生酒精奖赏的期望后, 不打算饮酒。因此, 该议定书涉及必要程度的欺骗。在述职时, 确保参与者知道他们不应该与其他人讨论协议。
虽然有证据表明, 这一程序有力地破坏了记忆, 并已被设计为最大限度地提高这种可能性, 但目前没有独立的方法来评估是否发生了不稳定。这是对迄今所描述的所有记忆失稳程序的限制, 因此, post-destabilization 干预在测试中的无效效果难以解释, 因为它们可能是由于干预效果低或未能充分首先破坏 MRMs 的稳定。
counterconditioning 效应对 non-trained 刺激的显著推广使这种方法在临床上具有很好的应用前景。然而, 我们的有效性演示仅限于一个实验性 (非) 设置与问题饮酒者, 谁没有酒精使用紊乱。
与, 虽然该程序很容易适应, 但不知道在使用人口的不同 (非法) 物质的破坏稳定 MRMs 的过程中, 是否采用同样有效的方法 (以修改后的形式)。我们认为, 这至少是有效的, 因为酒精 MRMs 可能是更 overlearned, 强健, 和 cross-contextual 比 MRMs 的大多数非法药物。然而, 这仍然有待于实验验证, 也不知道如何敏感的程序的功效是在数量的变化, 提出的线索或其确切的性质。因此, 我们鼓励在不同的药物使用人群中进行试验, 并采用不同的检索线索, 以及针对特定人群的奖励 maladaptations post-destabilization 干预。
为了解决上述测量问题, 即依赖 post-destabilization 干预的有效性来推断有效的记忆失稳-不稳定的独立措施是必需的。我们目前正在开发脑电图和心理技术来解决这一问题, 并将在完成后根据请求提供这些方法。
需要在酗酒的严重程度范围内实施这项技术的临床应用。此外, 临床研究是至关重要的, 以确定这一程序是否可以接受的治疗寻求者。

在刘易斯和同事的开创性工作上的建设1在过去二十年中, 通过内存再2、3的过程, 突出显示了在建立的 (整合的) 记忆中的可塑性空前的潜力。.再在某些情况下发生, 当记忆被检索4并且包括二个世俗地5和分子6分离阶段: 初始不稳定7和随后 restabilization8.在检索引起的记忆失稳之后, 记忆进入一个 ' 活跃 ', 可延展的状态, 在其中他们是易受修改通过新的信息的合并9,10,11,12, 或通过药理干扰减弱为 restabilization 所需的分子通路13、14、15、16。这 ' 再窗口, ' 在不稳定和 restabilization 之间的间隔, 持续1到 6 h 并且代表一个独特的机会直接地干预与适应奖赏记忆 (MRMs) 被相信在病因学中扮演一个关键的角色17, 进展18, 和维护19,20的物质使用紊乱 (肥皂泡沫), 如酒精使用紊乱。
MRMS 是通过巴甫洛夫机制形成的关联, 它们将环境刺激或 "药物线索" 与可用性联系起来, 以及药物的令人陶醉和奖励效果21。MRMs 灌输的药物提示与增强的价值, 突出22, 和激励性的属性, 使这些线索抓住注意23, 触发渴望和刺激药物寻求和使用的行为在人们的肥皂。因此, 这些 MRMs 被认为是负责触发复发, 当药物线索遇到24。因此, 对泡沫的长期临床管理的主要挑战是降低 MRMs 的能力, 以促进药物服用, 这, 通过延长, 应减少复发的发病率21。再窗口提供了一个激动人心的机会来实现这个目标, 更新 MRMs, 而他们不稳定的形式, 不太可能导致复发。
最近的研究表明, 这可以通过纯行为干预10,25实现。这些都能在海洛因成瘾者中产生强烈的渴求和药物相关的奖赏指标26, 吸烟者27, 和危险饮酒者28。通过简单地提醒 MRMs (通常是短暂接触药物提示) 来破坏他们的稳定, 并随后 re-associating 药物的线索, 缺乏药物 (灭绝), 或厌恶结局 (counterconditioning)29 , 而 MRMs 不稳定,药物提示和结果之间的关联让在一个更新的, 更适应的形式。这些重写 MRMs 不再支持复发。这些和标准灭绝或 counterconditioning 干预的唯一区别是在学习干预之前, 通过短暂接触药物线索而产生的 MRMs 的检索和不稳定。因此, 它们通常被称为 "检索-灭绝" 或 "检索-counterconditioning" 干预。
然而, 现在有一个重要的研究机构显示不一致的结果与这些干预30,31,32,33。这可能部份地解释由各自的区别在可塑性促进的蛋白质的表示34。但是, 由于检索是必需的, 但没有足够的4、35、36用于内存不稳定, 因此还有确定内存是否破坏的某些边界条件。这些边界条件的执行程度可能也因 study-to-study 而异。首要在这些情况之中是记忆年龄 (年代史)37和力量 (增强历史)38,39, 提醒试用5的持续时间, 以及不匹配的发生35,40在检索过程中的预测和实际结果之间, 称为预测错误 (PE)41、42、43。这些因素是相互关联的, 因为 PE 是中央奖励的 "学习信号"44,45 , 并在学习过程中减少。PE 对在内存中编码的预期结果与实际结果之间的差异进行编码, 反映了所学预测的不精确性。它是通过最小化发生的体育学习累算。老, 强烈加强记忆象 MRMs-已经形成在许多岁月-有非常好预测力量为结果横跨不同的情况。因此, 当在正常条件下检索 MRMs 时, 将产生很少的 PE, 从而赋予这些痕迹以不稳定的抵抗力。
利用这些知识, 我们开发了一个检索和 pe 生成 (检索 + pe) 的酒精 MRMs 的过程, 目的是最大限度地提高这些记忆的可能性, 通过最大化 PE 在检索的破坏。我们已经表明, 随后的 counterconditioning 导致全面更新酒精 MRMs 在危险饮酒者28,46。这篇手稿描述了执行的检索 + PE 程序的破坏性酒精 MRMs, 连同多 counterconditioning 技术, 重写这些记忆。
检索 + PE 程序涉及使用口头指示, 以最大限度地提高参与者的预期酒精奖励, 然后防止酒精消费在最后一刻。在没有确切的学习历史知识的情况下, 这种 PE 最大化认定最大化了 MRMs 失稳的可能性, 并提供了一个平台来测试 post-destabilization 行为干预对改善指数的有效性适应奖励记忆。在这里, 我们介绍如何实现此过程与后续的 disgust-based counterconditioning, 我们的28和其他29显示为更新 MRMs 的有效学习模式。然而, 检索 + PE 程序也可与替代 post-retrieval 药物干预和许多其他形式的学习干预措施结合使用25。

<table>
	<tbody>
		<tr>
			<td>Multipipette M4 x 1</td>
			<td>Eppendorf</td>
			<td>4982 000.012</td>
			<td>High-accuracy pipette for dosing Bitrex</td>
		</tr>
		<tr>
			<td>Combitips advanced 0.2ml x 1000</td>
			<td>Eppendorf</td>
			<td>0030 089.413</td>
			<td>Syringe tip for use with Micropipette for dosing Bitrex</td>
		</tr>
		<tr>
			<td>Simax 100ml measuring cylinder</td>
			<td>Phillip Harris</td>
			<td>B8A73325</td>
			<td>Any suitable measuring cylinder may be used</td>
		</tr>
		<tr>
			<td>Bitrex (2.5% w/v aqueaous Denatonium Benzoate solution)</td>
			<td>Mcfarlan Smith</td>
			<td>N/A</td>
			<td>Bitter solution for counterconditioning drink UCSs</td>
		</tr>
		<tr>
			<td>International Affective Picture System images</td>
			<td>International Affective Picture System (Lang et al, 2008)</td>
			<td>(http://csea.phhp.ufl.edu/media/iapsmessage.html)</td>
			<td>Upon request from Margaret Bradley&#160;</td>
		</tr>
		<tr>
			<td>Stimulus presentation monitor with 1024 x768 colour display minimum</td>
			<td>Dell&#160;</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Eyelink 1000 Core System</td>
			<td>SR Research</td>
			<td>Desktop-type mount</td>
			<td>Other eye-tracking systems can be used</td>
		</tr>
		<tr>
			<td>Experiment Builder</td>
			<td>SR Resesarch</td>
			<td>N/A</td>
			<td>Other systems (Presentation/MATLAB/ePrime/Psychopy etc) can be used</td>
		</tr>
		<tr>
			<td>DataViewer</td>
			<td>SR Resesarch</td>
			<td>N/A</td>
			<td>Use software as appropriate to eye tracker setup</td>
		</tr>
		<tr>
			<td>SPSS 21.0 for Windows</td>
			<td>IBM</td>
			<td>Base Package</td>
			<td>Use any appopriate statistical software</td>
		</tr>
		<tr>
			<td>Lion Alcometer 500</td>
			<td>Lion Laboratories</td>
			<td>N/A</td>
			<td>Any accurate alcohol breath testing device may be used</td>
		</tr>
		<tr>
			<td>Terumo 5ml syringe</td>
			<td>MediSupplies</td>
			<td>PMCO595</td>
			<td>Any 5ml syringe may be used</td>
		</tr>
		<tr>
			<td>Mini Table Top Scale</td>
			<td>On Balance</td>
			<td>MTT-200</td>
			<td>Any high-accuracy scales may be used</td>
		</tr>
		<tr>
			<td>Non-alcoholic beer</td>
			<td>Bavaria</td>
			<td>N/A</td>
			<td>Any brand non-acloholic beer may be used</td>
		</tr>
	</tbody>
</table>

a prediction error-driven retrieval procedure for destabilizing and rewriting maladaptive reward memories in hazardous drinkers

适应奖励记忆 (MRMs) 可能会变得不稳定后, 在一定条件下检索, 允许他们修改后的新学习。然而, 健壮的 (排练) 和按年代排列的老 MRMs, 例如那些潜在的物质使用障碍, 在检索时不会容易地动摇。在检索过程中, 记忆失稳的关键是预测误差 (PE)。我们描述了一个在危险饮酒者中的酒精 MRMs 的检索程序, 它特别旨在最大限度地增加 PE 的生成, 因此可能造成 MRM 不稳定。该程序要求显着产生酒精消费量的预期, 然后违反这一预期 (预扣酒精) 后, 介绍了一套简短的原型酒精提示图像 (检索 + PE)。控制程序涉及呈现相同的提示图像, 但允许酒精被消耗, 产生最小的 pe (检索-无 pe) 或生成 pe 不回收酒精 MRMs, 通过提出橙汁提示 (没有检索 + pe)。随后, 我们描述了一个多 disgust-based counterconditioning 程序, 以探测在再之前重写酒精提示奖励协会的 MRM 不稳定。这一过程对酒精提示与图像援引病原体厌恶和极端苦味的解决方案 (铵苯甲酸酯), 产生味觉厌恶。以下检索 + pe, 但不是没有检索 + pe 或检索-没有 pe, counterconditioning 产生的证据, MRM 重写作为索引的持久减少酒精提示估价, 注意捕获, 和酒精渴求。

比较组指在基线上的主观测度。组应等同于基线摄入酒精, 厌恶敏感性/倾向, 苏格拉底和 NAEQ 评分, 和一时的渴望 (ACQ 现在)。如果这些措施在基线上存在差异, 则应继续使用混合效果模型进行分析, 并将这些措施作为随机效果59。这里没有描述这些分析的性能。评估每组阳性酒精酒精读数的频率, 并报告结果。
cs + s 和 cs s 的愉悦度在基线上没有差异 (在 counterconditioning 之前), 但对 counterconditioning 的试验分岔, 随着对啤酒 cs 的愉悦评分, 通过与画报的反复配对而显著降低。苦饮 UCSs。如果 cs + s 和 cs s 在基线上不同, 请使用随机拦截和斜率的混合效果模型来分析这些数据。CS-s 从来没有配对的 UCSs, 所以不要表现出下降的愉悦。这由 cs 类型 (cs + vs. cs) x 试验 (基线、试验1、2、3 & 4) 相互作用 (如果 counterconditioning 已成功), 如图 2所示 (Das et al.中的代表数据)28)
重写 MRM 网络的证据是通过对酒精刺激的愉悦度进行广义的降低来提供的, 而不是在后续的中性刺激 (8 天)。在零售 + PE 组, 减少的愉悦度是明显的响应 cs + s 使用的检索/counterconditioning 任务 (啤酒 cs + s), 以及新的啤酒和葡萄酒的图像评级第一次跟进。统计学上, 这是明显的 cs 类型 (啤酒 cs +, 中性 cs-, 新颖的啤酒, 新颖的葡萄酒) x 组 (无零售 + pe, 零售 + pe, 没有 pe) 的互动。示例结果显示此模式取自 Das et al.28显示在图 3中。
在可选的注意力偏倚任务中检查结果的模式。注意偏差被计算为在目标的酒精图像上停留时间在它的组成匹配非靶对图像上停留时间。评分为0表示没有偏见, 积极的得分注意偏向酒精刺激, 和一个负评分对酒精图片的动眼神经反感。取自 Das et al.的代表性结果28显示在图 4中。可以看出, 对酒精图像在后续 (天 8) 的动眼厌恶只产生于零售 + PE 组, 由一个显着的组 x CS 类型的互动表示。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56097/56097fig1.jpg"> 图 1: 三检索组的示意图和预测错误程序。 缩写的关键:零售 + PE = 检索 + 预测错误;该组谁检索酒精的记忆, 并期待酒精扣留。没有 PE = 检索 + 无预测错误;该组的人回收酒精的记忆和消费酒精的预期。无零售+ PE = 无检索 + 预测错误;该组不回收酒精的记忆, 并期待橙汁扣留。在体内饮料: 150 毫升的啤酒在零售 + pe 和零售 + 没有 pe 或150毫升橙汁玻璃在没有零售组。1) 在体内饮料:与 particpants 组相关的饮料放在参与者前面, 他们被告知他们将会根据屏幕上的提示饮用这种饮料;后, 一系列的图片为愉悦。2)提示图像分级:参加者 + pe 和零售 + 没有 pe 率四图像的啤酒和两个图像的软饮料 (咖啡和可乐) 的愉悦。简单的在体内暴露于啤酒和啤酒的图片评级会触发这些群体中饮酒记忆的检索。No 的零售组率四图像橙汁和软饮料图像的愉悦, 因此不回收酒精的记忆。3)预测错误:由于所有参与者都被告知, 他们将在程序开始时饮用 "体内" 饮料, pe 是通过防止参加者在其前面的饮料中产生的, 而不是在其上使用。在没有 pe 的时候, 饮料按预期消耗, 不产生 pe。4) 干扰项: 执行高工作记忆任务 (散文召回、数字跨度、类别和语言流畅), 以分散参与者的注意力并使他们脱离检索任务。5) 干预: 一个 counterconditioning 的任务, 其中啤酒图像与令人厌恶的图像和非常苦涩的饮料配对。这一过程的目的是建立一个啤酒的形象和厌恶的经验之间的联系, 覆盖现有的协会, 由于酒精记忆的不稳定。<a href="//cloudflare.jove.com/files/ftp_upload/56097/56097fig1large.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/56097/56097fig2.jpg"> 图 2: counterconditioning 期间的中性和啤酒提示的平均愉悦度 (± SEM).在 counterconditioning 过程中, 从 Das et al.中, cs + s (啤酒图像与恶心结果配对) 和 cs s (中性图像配对中性结局) 的愉悦度的代表性变化28作为 counterconditioning 的目的是援引有条件的厌恶, 以响应啤酒的图像, 感知愉悦的这些图像应减少在 counterconditioning。评级在基线上没有差别, 但在 counterconditioning 的过程中却减少到 CS。** = cs + > cs-在p< 0.001。<a href="//cloudflare.jove.com/files/ftp_upload/56097/56097fig2large.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/56097/56097fig3.jpg"> 图 3: 平均愉悦度 (± SEM) 的 counterconditioned, 新的酒精提示和中性提示在测试.代表普遍降低酒精图像的愉悦度 (喜欢) 七天后, counterconditioning 是执行以下检索 (零售) 与预测错误 (pe) 从Das et al。28此模式与对与酒精有关的奖励协会的重写是一致的。键:中性 CS- = 在 counterconditioning 期间与中性图像配对的咖啡和可乐图像。啤酒 CS + = counterconditioning 期间的啤酒图像与恶心的结果配对。新奇啤酒= 以前看不见的啤酒图片。葡萄酒= previousy 看不见的葡萄酒图像。* = 在p< 0.05 上的显著配对比较, Bonferroni 更正。<a href="//cloudflare.jove.com/files/ftp_upload/56097/56097fig3large.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/56097/56097fig4.jpg"> 图 4: 对 counterconditioned 的平均注意偏向、新的酒精提示和测试中的中性提示.代表普遍取消注意偏见的所有酒精刺激组的 counterconditioning 后, 从Das et al28。酒吧表示, 平均注意偏倚评分, 计算为总时间的平均差异, 看一个 ' 目标图像 ' 含有可见的酒精与组合匹配的 ' 非目标 ' imagelacking 可见酒精。误差线是中小企业。中性 CS- = 在 counterconditioning 期间与中性图像配对的咖啡和可乐图像。啤酒 CS + = counterconditioning 期间的啤酒图像与恶心的结果配对。新奇啤酒= 以前看不见的啤酒图片。葡萄酒= 以前看不见的葡萄酒图片。* = 在p< 0.05 上的显著配对比较, Bonferroni 更正。<a href="//cloudflare.jove.com/files/ftp_upload/56097/56097fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td>试用编号</td>
			<td colspan="2">CS 型</td>
			<td colspan="2">UCS 类型</td>
			<td colspan="2">UCS 文件名</td>
		</tr>
<tr>
<td>1</td>
			<td colspan="2">啤酒 CS+1</td>
			<td colspan="2">味觉</td>
			<td colspan="2">DRINK_SCREENpng</td>
		</tr>
<tr>
<td>2</td>
			<td colspan="2">中性 CS-1</td>
			<td colspan="2">中性</td>
			<td colspan="2">7020.jpg</td>
		</tr>
<tr>
<td>3</td>
			<td colspan="2">啤酒 CS+1</td>
			<td colspan="2">味觉</td>
			<td colspan="2">DRINK_SCREENpng</td>
		</tr>
<tr>
<td>4</td>
			<td colspan="2">中性 CS-2</td>
			<td colspan="2">中性</td>
			<td colspan="2">7019.jpg</td>
		</tr>
<tr>
<td>5</td>
			<td colspan="2">啤酒 CS+4</td>
			<td colspan="2">图案</td>
			<td colspan="2">rotten_foot. jpg</td>
		</tr>
<tr>
<td>6</td>
			<td colspan="2">啤酒 CS+3</td>
			<td colspan="2">图案</td>
			<td colspan="2">9325.jpg</td>
		</tr>
<tr>
<td>7</td>
			<td colspan="2">啤酒 CS+2</td>
			<td colspan="2">味觉</td>
			<td colspan="2">DRINK_SCREENpng</td>
		</tr>
<tr>
<td>8</td>
			<td colspan="2">啤酒 CS+4</td>
			<td colspan="2">图案</td>
			<td colspan="2">9325.jpg</td>
		</tr>
<tr>
<td>9</td>
			<td colspan="2">啤酒 CS+3</td>
			<td colspan="2">图案</td>
			<td colspan="2">rotten_foot. jpg</td>
		</tr>
<tr>
<td>10</td>
			<td colspan="2">中性 CS-1</td>
			<td colspan="2">中性</td>
			<td colspan="2">7020.jpg</td>
		</tr>
<tr>
<td>11</td>
			<td colspan="2">啤酒 CS+1</td>
			<td colspan="2">味觉</td>
			<td colspan="2">DRINK_SCREENpng</td>
		</tr>
<tr>
<td>12</td>
			<td colspan="2">中性 CS-2</td>
			<td colspan="2">中性</td>
			<td colspan="2">7019.jpg</td>
		</tr>
<tr>
<td>13</td>
			<td colspan="2">中性 CS-1</td>
			<td colspan="2">中性</td>
			<td colspan="2">7019.jpg</td>
		</tr>
<tr>
<td>14</td>
			<td colspan="2">啤酒 CS+2</td>
			<td colspan="2">味觉</td>
			<td colspan="2">DRINK_SCREENpng</td>
		</tr>
<tr>
<td>15</td>
			<td colspan="2">啤酒 CS+4</td>
			<td colspan="2">图案</td>
			<td colspan="2">9405.jpg</td>
		</tr>
<tr>
<td>16</td>
			<td colspan="2">啤酒 CS+1</td>
			<td colspan="2">味觉</td>
			<td colspan="2">DRINK_SCREENpng</td>
		</tr>
<tr>
<td>17</td>
			<td colspan="2">啤酒 CS+4</td>
			<td colspan="2">图案</td>
			<td colspan="2">9301.jpg</td>
		</tr>
<tr>
<td>18</td>
			<td colspan="2">中性 CS-2</td>
			<td colspan="2">中性</td>
			<td colspan="2">7019.jpg</td>
		</tr>
<tr>
<td>19</td>
			<td colspan="2">啤酒 CS+3</td>
			<td colspan="2">图案</td>
			<td colspan="2">9405.jpg</td>
		</tr>
<tr>
<td>20</td>
			<td colspan="2">啤酒 CS+2</td>
			<td colspan="2">味觉</td>
			<td colspan="2">DRINK_SCREENpng</td>
		</tr>
<tr>
<td>21</td>
			<td colspan="2">啤酒 CS+3</td>
			<td colspan="2">图案</td>
			<td colspan="2">9301.jpg</td>
		</tr>
<tr>
<td>22</td>
			<td colspan="2">中性 CS-2</td>
			<td colspan="2">中性</td>
			<td colspan="2">7019.jpg</td>
		</tr>
<tr>
<td>23</td>
			<td colspan="2">啤酒 CS+2</td>
			<td colspan="2">味觉</td>
			<td colspan="2">DRINK_SCREENpng</td>
		</tr>
<tr>
<td>24</td>
			<td colspan="2">中性 CS-1</td>
			<td colspan="2">中性</td>
			<td colspan="2">7020.jpg</td>
		</tr>
</tbody></table>表 1: Pseudorandomized 在 Das 中使用的 counterconditioning 任务的试用顺序et al. 28 CS 类型= 条件刺激类型。counterconditioning 有四啤酒的图片。这些是任意编号的啤酒 CS + 1 到4。两个中性的咖啡和可乐的图像也使用, 这些都是任意编号的中性 CS-1 和中性 CS-2。UCS 类型= 无条件刺激类型。味觉无条件刺激是 "现在喝" 出现的 on-screen, 参与者消耗15毫升0.067% 铵苯甲酸溶液。画报UCSs 是来自国际情感图片系统 (IAPS) 数据库和互联网的令人不快/恶心的图像。中性UCSs 是两个图像来源的 IAPS 数据库, 是额定中立。UCS 文件名:此列提供在每个试验中显示为结果的特定图像。数字文件名指的是 IAPS 的数据库号。这些图像可在请求 IAPS 数据库时使用。其他图像可从作者要求。

Watch this Scientific Journal Video about A Prediction Error-driven Retrieval Procedure for Destabilizing and Rewriting Maladaptive Reward Memories in Hazardous Drinkers at JoVE.com

A Prediction Error-driven Retrieval Procedure for Destabilizing and Rewriting Maladaptive Reward Memories in Hazardous Drinkers

这份手稿描述了一个记忆检索程序, 破坏稳定的奖励记忆和重写他们的 counterconditioning 之前, 他们的再。

危险饮酒者适应奖赏记忆的预测误差驱动检索方法

Maladaptive reward memories (MRMs) can become unstable following retrieval under certain conditions, allowing their modification by subsequent new ...

a-prediction-error-driven-retrieval-procedure-for-destabilizing

Research

JoVE Journal

Neuroscience

9.7K 次观看.  University College London. 这份手稿描述了一个记忆检索程序, 破坏稳定的奖励记忆和重写他们的 counterconditioning 之前, 他们的再。

视频: A Prediction Error-driven Retrieval Procedure for Destabilizing and Rewriting Maladaptive Reward Memories in Hazardous Drinkers

SDS-PAGE/Immunoblot Detection of A&beta; Multimers in Human Cortical Tissue Homogenates using Antigen-Epitope Retrieval

The anomalous folding and polymerization of the &beta;-amyloid (A&beta;) peptide is thought to initiate the neurodegenerative cascade in Alzheimer's disease pathogenesis1. A&beta; is predominantly a 40- or 42-amino acid peptide that is prone to self-aggregation into &beta;-sheet-rich amyloid fibrils that are found in the cores of cerebral senile plaques in Alzheimer's disease. Increasing evidence suggests that low molecular weight, soluble A&beta; multimers are more toxic than fibrillar A&beta; amyloid2. The identification and quantification of low- and high-molecular weight multimeric A&beta; species in brain tissue is an essential objective in Alzheimer's disease research, and the methods employed also can be applied to the identification and characterization of toxic multimers in other proteopathies3. Naturally occurring A&beta; multimers can be detected by SDS-polyacrylamide gel electrophoresis followed by immunoblotting with A&beta;-specific antibodies. However, the separation and detection of multimeric A&beta; requires the use of highly concentrated cortical homogenates and antigen retrieval in small pore-size nitrocellulose membranes. Here we describe a technique for the preparation of clarified human cortical homogenates, separation of proteins by SDS-PAGE, and antigen-epitope retrieval/Western blotting with antibody 6E10 to the N-terminal region of the A&beta; peptide. Using this protocol, we consistently detect A&beta; monomers, dimers, trimers, tetramers, and higher molecular weight multimers in cortical tissue from humans with Alzheimer's pathology.

SDS-PAGE/Immunoblot Detection of A&#946; Multimers in Human Cortical Tissue Homogenates using Antigen-Epitope Retrieval

We describe a technique for the preparation of clarified human cortical homogenates, protein separation by SDS-PAGE, antigen retrieval and immunoblotting with an antibody to the A&beta; peptide. Using this protocol, we consistently detect monomeric and multimeric A&beta; in cortical tissue from humans with Alzheimer's pathology.

SDS-PAGE/Immunoblot检测人脑皮层组织中的Aβ多聚匀浆使用抗原表位检索

The anomalous folding and polymerization of the &beta;-amyloid (A&beta;) peptide is thought to initiate the neurodegenerative cascade in Alzheimer's ...

科研

神经科学

A Procedure for Studying the Footshock-Induced Reinstatement of Cocaine Seeking in Laboratory Rats

The most insidious aspect of drug addiction is the high propensity for relapse. Animal models of relapse, known as reinstatement procedures, have been used extensively to study the neurobiology and phenomenology of relapse to drug use. Although procedural variations have emerged over the past several decades, the most conventional reinstatement procedures are based on the drug self-administration (SA) model. In this model, an animal is trained to perform an operant response to obtain drug. Subsequently, the behavior is extinguished by withholding response-contingent reinforcement. Reinstatement of drug seeking is then triggered by a discrete event, such as an injection of the training drug, re-exposure to drug-associated cues, or exposure to a stressor 1.
Reinstatement procedures were originally developed to study the ability of acute non-contingent exposure to the training drug to reinstate drug seeking in rats and monkeys 1, 2. Reinstatement procedures have since been modified to study the role of environmental stimuli, including drug-associated cues and exposure to various forms of stress, in relapse to drug seeking 1, 3, 4.
Over the past 15 years, a major focus of the reinstatement literature has been on the role of stress in drug relapse. One of the most commonly used forms of stress for studying this relationship is acute exposures to mild, intermittent, electric footshocks. The ability of footshock stress to induce reinstatement of drug seeking was originally demonstrated by Shaham and colleagues (1995) in rats with a history of intravenous heroin SA5. Subsequently, the effect was generalized to rats with histories of intravenous cocaine, methamphetamine, and nicotine SA, as well as oral ethanol SA 3, 6.
Although footshock-induced reinstatement of drug seeking can be achieved reliably and robustly, it is an effect that tends to be sensitive to certain parametrical variables. These include the arrangement of extinction and reinstatement test sessions, the intensity and duration of footshock stress, and the presence of drug-associated cues during extinction and testing for reinstatement. Here we present a protocol for footshock-induced reinstatement of cocaine seeking that we have used with consistent success to study the relationship between stress and cocaine seeking.

Animal models of relapse, known as reinstatement procedures, have been used extensively to study the role of stress in relapse to drug seeking. Here, we report on a method for inducing the reinstatement of cocaine seeking in laboratory rats via acute exposures to mild, intermittent electric footshock.

在实验室老鼠研究求可卡因电击诱导恢复原状的程序

The most insidious aspect of drug addiction is the high propensity for relapse.  Animal models of relapse, known as reinstatement procedures, have been ...

A General Method for Evaluating Incubation of Sucrose Craving in Rats

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and relapse to food taking 1. Food craving conceptualized in this way is akin to drug craving in response to drug-paired cues. A rich literature has been developed around understanding the behavioral and neurobiological determinants of drug craving; we and others have been focusing recently on translating techniques from basic addiction research to better understand addiction-like behaviors related to food 2-4.
As done in previous studies of drug craving, we examine sucrose craving behavior by utilizing a rat model of relapse. In this model, rats self-administer either drug or food in sessions over several days. In a session, lever responding delivers the reward along with a tone+light stimulus. Craving behavior is then operationally defined as responding in a subsequent session where the reward is not available. Rats will reliably respond for the tone+light stimulus, likely due to its acquired conditioned reinforcing properties 5. This behavior is sometimes referred to as sucrose seeking or cue reactivity. In the present discussion we will use the term "sucrose craving" to subsume both of these constructs.
In the past decade, we have focused on how the length of time following reward self-administration influences reward craving. Interestingly, rats increase responding for the reward-paired cue over the course of several weeks of a period of forced-abstinence. This "incubation of craving" is observed in rats that have self-administered either food or drugs of abuse 4,6. This time-dependent increase in craving we have identified in the animal model may have great potential relevance to human drug and food addiction behaviors.
Here we present a protocol for assessing incubation of sucrose craving in rats. Variants of the procedure will be indicated where craving is assessed as responding for a discrete sucrose-paired cue following extinction of lever pressing within the sucrose self-administration context (Extinction without cues) or as responding for sucrose-paired cues in a general extinction context (Extinction with cues).

Responding for food or drug-paired cues increases over the course of a period of abstinence and this may relate to an increased susceptibility to relapse behaviors. Here we detail a procedure for evaluating this "incubation of craving" in rats that have self-administered sucrose.

评估大鼠蔗糖渴求育成的一个通用方法

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and ...

A Lightweight, Headphones-based System for Manipulating Auditory Feedback in Songbirds

Experimental manipulations of sensory feedback during complex behavior have provided valuable insights into the computations underlying motor control and sensorimotor plasticity1. Consistent sensory perturbations result in compensatory changes in motor output, reflecting changes in feedforward motor control that reduce the experienced feedback error. By quantifying how different sensory feedback errors affect human behavior, prior studies have explored how visual signals are used to recalibrate arm movements2,3 and auditory feedback is used to modify speech production4-7. The strength of this approach rests on the ability to mimic naturalistic errors in behavior, allowing the experimenter to observe how experienced errors in production are used to recalibrate motor output.
Songbirds provide an excellent animal model for investigating the neural basis of sensorimotor control and plasticity8,9. The songbird brain provides a well-defined circuit in which the areas necessary for song learning are spatially separated from those required for song production, and neural recording and lesion studies have made significant advances in understanding how different brain areas contribute to vocal behavior9-12. However, the lack of a naturalistic error-correction paradigm - in which a known acoustic parameter is perturbed by the experimenter and then corrected by the songbird - has made it difficult to understand the computations underlying vocal learning or how different elements of the neural circuit contribute to the correction of vocal errors13.
The technique described here gives the experimenter precise control over auditory feedback errors in singing birds, allowing the introduction of arbitrary sensory errors that can be used to drive vocal learning. Online sound-processing equipment is used to introduce a known perturbation to the acoustics of song, and a miniaturized headphones apparatus is used to replace a songbird's natural auditory feedback with the perturbed signal in real time. We have used this paradigm to perturb the fundamental frequency (pitch) of auditory feedback in adult songbirds, providing the first demonstration that adult birds maintain vocal performance using error correction14. The present protocol can be used to implement a wide range of sensory feedback perturbations (including but not limited to pitch shifts) to investigate the computational and neurophysiological basis of vocal learning.

We describe the design and assembly of miniaturized headphones suitable for replacing a songbird&#x2019;s natural auditory feedback with a manipulated acoustic signal. Online sound processing hardware is used to manipulate song output, introduce real-time errors in auditory feedback via the headphones, and drive vocal motor learning.

一个轻量级的，耳机为基础的系统操作鸣禽听觉反馈

Experimental manipulations of sensory feedback during complex behavior have provided valuable insights into the computations underlying motor control and ...

A Procedure for Implanting Organized Arrays of Microwires for Single-unit Recordings in Awake, Behaving Animals

In vivo electrophysiological recordings in the awake, behaving animal provide a powerful method for understanding neural signaling at the single-cell level. The technique allows experimenters to examine temporally and regionally specific firing patterns in order to correlate recorded action potentials with ongoing behavior. Moreover, single-unit recordings can be combined with a plethora of other techniques in order to produce comprehensive explanations of neural function. In this article, we describe the anesthesia and preparation for microwire implantation. Subsequently, we enumerate the necessary equipment and surgical steps to accurately insert a microwire array into a target structure. Lastly, we briefly describe the equipment used to record from each individual electrode in the array. The fixed microwire arrays described are well-suited for chronic implantation and allow for longitudinal recordings of neural data in almost any behavioral preparation. We discuss tracing electrode tracks to triangulate microwire positions as well as ways to combine microwire implantation with immunohistochemical techniques in order to increase the anatomical specificity of recorded results.

Implanting organized arrays of microwires for use in single-unit electrophysiological recordings presents a number of technical challenges.&#160;Methods for performing this technique and the equipment necessary are described.&#160;Also, the beneficial use of organized microwire arrays to record from distinct neural subregions with high spatial selectivity is discussed.

一个程序植入微丝的有组织阵列的单机录音清醒，举止动物

In vivo electrophysiological recordings in the awake, behaving animal provide a powerful method for understanding neural signaling at the single-cell ...

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by repeated, time-lapse in vivo imaging in live rodents. These studies were made possible by straightforward surgical procedures, which enabled optical access for a prolonged period of time without repeat surgical procedures. In contrast, analogous studies of the spinal cord have been previously limited to only a few imaging sessions, each of which required an invasive surgery. As previously described, we have developed a spinal chamber that enables continuous optical access for upwards of 8 weeks, preserves mechanical stability of the spinal column, is easily stabilized externally during imaging, and requires only a single surgery. Here, the design of the spinal chamber with its associated surgical implements is reviewed and the surgical procedure is demonstrated in detail. Briefly, this video will demonstrate the preparation of the surgical area and mouse for surgery, exposure of the spinal vertebra and appropriate tissue debridement, the delivery of the implant and vertebral clamping, the completion of the chamber, the removal of the delivery system, sealing of the skin, and finally, post-operative care. The procedure for chronic in vivo imaging using nonlinear microscopy will also be demonstrated. Finally, outcomes, limitations, typical variability, and a guide for troubleshooting are discussed.

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

In this video, we describe a procedure for implanting a chronic optical imaging chamber over the dorsal spinal cord of a live mouse. The chamber, surgical procedure, and chronic imaging are reviewed and demonstrated.

一个程序，植入脊柱商会纵<em&gt;在体内</em&gt;鼠标脊髓成像

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by ...

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life. The cold plantar assay allows the objective and inexpensive assessment of cold sensitivity in mice, and can quantify both analgesia and hypersensitivity. Mice are acclimated on a glass plate, and a compressed dry ice pellet is held against the glass surface underneath the hindpaw. The latency to withdrawal from the cooling glass is used as a measure of cold sensitivity.
Cold sensation is also important for survival in regions with seasonal temperature shifts, and in order to maintain sensitivity animals must be able to adjust their thermal response thresholds to match the ambient temperature. The Cold Plantar Assay (CPA) also allows the study of adaptation to changes in ambient temperature by testing the cold sensitivity of mice at temperatures ranging from 30 &#176;C to 5 &#176;C. Mice are acclimated as described above, but the glass plate is cooled to the desired starting temperature using aluminum boxes (or aluminum foil packets) filled with hot water, wet ice, or dry ice. The temperature of the plate is measured at the center using a filament T-type thermocouple probe. Once the plate has reached the desired starting temperature, the animals are tested as described above.
This assay allows testing of mice at temperatures ranging from innocuous to noxious. The CPA yields unambiguous and consistent behavioral responses in uninjured mice and can be used to quantify both hypersensitivity and analgesia. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

The Cold Plantar Assay (CPA) measures cold responsiveness between 30 &#176;C and 5 &#176;C, and can also measure cold adaptation. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

测定冷灵敏度和适应小鼠一种简单廉价的方法

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

Polygraphic Recording Procedure for Measuring Sleep in Mice

Recording of the epidural electroencephalogram (EEG) and electromyogram (EMG) in small animals, like mice and rats, has been pivotal to study the homeodynamics and circuitry of sleep-wake regulation. In many laboratories, a cable-based sleep recording system is used to monitor the EEG and EMG in freely behaving mice in combination with computer software for automatic scoring of the vigilance states on the basis of power spectrum analysis of EEG data. A description of this system is detailed herein. Steel screws are implanted over the frontal cortical area and the parietal area of 1 hemisphere for monitoring EEG signals. In addition, EMG activity is monitored by the bilateral placement of wires in both neck muscles. Non-rapid eye movement (Non-REM; NREM) sleep is characterized by large, slow brain waves with delta activity below 4 Hz in the EEG, whereas a shift from low-frequency delta activity to a rapid low-voltage EEG in the theta range between 6 and 10 Hz can be observed at the transition from NREM to REM sleep. By contrast, wakefulness is identified by low- to moderate-voltage brain waves in the EEG trace and significant EMG activity.

The recording of electroencephalogram (EEG) and electromyogram (EMG) in freely behaving mice is a critical step to correlate behavior and physiology with sleep and wakefulness. The experimental protocol described herein provides a cable-based system for acquiring EEG and EMG recordings in mice.

印刷业记录程序在小鼠体内测量睡眠

Recording of the epidural electroencephalogram (EEG) and electromyogram (EMG) in small animals, like mice and rats, has been pivotal to study the ...

Simultaneous Detection of c-Fos Activation from Mesolimbic and Mesocortical Dopamine Reward Sites Following Naive Sugar and Fat Ingestion in Rats

This study uses cellular c-fos activation to assess effects of novel ingestion of fat and sugar on brain dopamine (DA) pathways in rats. Intakes of sugars and fats are mediated by their innate attractions as well as learned preferences. Brain dopamine, especially meso-limbic and meso-cortical projections from the ventral tegmental area (VTA), has been implicated in both of these unlearned and learned responses. The concept of distributed brain networks, wherein several sites and transmitter/peptide systems interact, has been proposed to mediate palatable food intake, but there is limited evidence empirically demonstrating such actions. Thus, sugar intake elicits DA release and increases c-fos-like immunoreactivity (FLI) from individual VTA DA projection zones including the nucleus accumbens (NAC), amygdala (AMY) and medial prefrontal cortex (mPFC) as well as the dorsal striatum. Further, central administration of selective DA receptor antagonists into these sites differentially reduce acquisition and expression of conditioned flavor preferences elicited by sugars or fats. One approach by which to determine whether these sites interacted as a distributed brain network in response to sugar or fat intake would be to simultaneous evaluate whether the VTA and its major mesotelencephalic DA projection zones (prelimbic and infralimbic mPFC, core and shell of the NAc, basolateral and central-cortico-medial AMY) as well as the dorsal striatum would display coordinated and simultaneous FLI activation after oral, unconditioned intake of corn oil (3.5%), glucose (8%), fructose (8%) and saccharin (0.2%) solutions. This approach is a successful first step in identifying the feasibility of using cellular c-fos activation simultaneously across relevant brain sites to study reward-related learning in ingestion of palatable food in rodents.

The goal of this study is to identify reward-related distributed brain networks by delineating a reliable immunohistological technique using cellular c-fos activation to measure simultaneous changes in dopamine pathways and terminal sites after novel ingestion of fat and sugar in rats.

从脑边缘和Mesocortical多巴胺奖赏网站继朴素的糖和脂肪摄入大鼠c-fos激活的同时检测

This study uses cellular c-fos activation to assess effects of novel ingestion of fat and sugar on brain dopamine (DA) pathways in rats. Intakes of sugars ...

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We demonstrate a simple way to tap into the fly reward system, modify experiences related to natural reward, and use voluntary ethanol consumption as a measure for changes in reward states. The approach serves as a relevant tool to study the neurons and genes that play a role in experience-mediated changes of internal state. The protocol is composed of two discrete parts: exposing the flies to rewarding and nonrewarding experiences, and assaying voluntary ethanol consumption as a measure of the motivation to obtain a drug reward. The two parts can be used independently to induce the modulation of experience as an initial step for further downstream assays or as an independent two-choice feeding assay, respectively. The protocol does not require a complicated setup and can therefore be applied in any laboratory with basic fly culture tools.

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

We describe a protocol for inducing rewarding and nonrewarding experiences in fruit flies (Drosophila melanogaster) using voluntary ethanol consumption as a measure for changes in reward states.

一个简单的方法来衡量在改建悬赏寻求行为使用<em&gt;果蝇</em

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We ...

危险饮酒者适应奖赏记忆的预测误差驱动检索方法

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