通过嗅觉挖掘任务评估小鼠判断偏差
1Department of Integrative Biology,University of Guelph, 2Institute of Cell Biology and Neurosciences,National Scientific and Technical Research Council-University of Buenos Aires, 3Laboratory of Experimental Animals, Faculty of Veterinary Sciences,National University of La Plata, 4Department of Animal Biosciences,University of Guelph
Summary
本文详细介绍了一种新型小鼠判断偏倚协议。还证明了这种嗅觉挖掘任务对情感状态的敏感性的证据，并讨论了其在不同研究领域的实用性。
Abstract
判断偏差（JB）是处于积极和消极情感/情绪状态的个体解释模棱两可信息的方式的差异。这种现象在人类中早已观察到，处于积极状态的个体“乐观地”对模糊性做出反应，而处于消极状态的个体则表现出“悲观主义”。旨在评估动物影响的研究人员利用了这些差异反应，开发了评估判断偏差的任务，作为情感状态的指标。这些任务在各种物种和研究领域越来越受欢迎。然而，对于实验室小鼠，研究中使用最广泛的脊椎动物和严重依赖的物种来模拟情感障碍，只有一个JB任务被成功验证为对情感状态的变化敏感。在这里，我们详细介绍了这种新型小鼠JB任务，并证明了它对小鼠的影响是敏感的。虽然改进仍然是必要的，但对小鼠JB的评估为回答有关小鼠福利的实际问题和关于情感状态在转化研究中的影响的基本问题打开了大门。
Introduction
测量情感调节的判断偏差（以下简称JB）已被证明是研究动物情绪状态的有用工具。这种创新方法借鉴了人类心理学，因为经历积极或消极情感状态（情绪和长期情绪）的人类可靠地证明了他们处理信息的方式的差异1，2，3。例如，经历焦虑或抑郁的人可能会将中性的面部表情解释为消极的，或将中性的句子解释为威胁4，5。这些偏差很可能具有适应性值，因此在物种6，7中是保守的。旨在评估动物影响的研究人员巧妙地利用了这一现象，将乐观主义视为对中性或模糊线索的奖励期望增加，并将悲观主义视为对惩罚或奖励缺失的期望增加8，9。因此，在实验环境中，对模糊刺激的乐观和悲观反应可以分别解释为积极和消极影响的指标，分别为10，11。
与其他动物影响指标相比，JB任务有可能成为特别有价值的工具，因为它们能够检测情感状态的价态和强度10，11。JB任务检测阳性状态的能力（例如，Rygula等人12）特别有用，因为大多数动物影响指标仅限于检测阴性状态13。在JB任务期间，动物通常被训练对预测奖励的正判别提示（例如，高频音调）和预测惩罚的负判别提示（例如，低频音调）做出反应，然后再呈现模棱两可的提示（例如， 中间音调）8。如果为了回应模棱两可的线索，动物“乐观地”对积极线索执行训练的反应（好像期待奖励），这表明存在积极的判断偏差。或者，如果动物表现出消极的训练反应以避免惩罚，这表明“悲观主义”或消极的判断偏差。
自从哈丁及其同事8成功开发出第一个动物JB任务以来，已经为不同研究领域的各种物种开发了几个JB任务7。但是，尽管它们越来越受欢迎，但动物JB任务通常是劳动密集型的。此外，也许是因为它们在方法上与启发它们的人工任务不同，它们有时会产生无效或违反直觉的结果14 ，并且通常只产生很小的治疗效果大小15。因此，开发和实现 JB 任务可能具有挑战性。事实上，对于实验室小鼠，研究中使用最广泛的脊椎动物16，17 和严重依赖的物种来模拟情感障碍18，尽管在过去十年中进行了许多尝试，但只有一项JB任务被成功验证为对情感状态变化敏感19 （参见Resasco等人的补充材料19 以获取摘要）。本文描述了最近验证的murine JB任务，详细介绍了其生物学上的相关设计，并强调了将这种人道任务应用于测试与小鼠影响相关的重要假设的方式。总体而言，该协议可以用于研究小鼠JB上任何感兴趣的变量的情感效应。这将包括此处描述的分类治疗变量（药物或疾病影响，环境条件，遗传背景等），或与连续变量的关系（生理变化，家庭笼子行为等）。
Protocol
实验由圭尔夫大学动物护理委员会（AUP #3700）批准，符合加拿大动物护理委员会指南，并根据ARREAT（动物研究： 体内 实验报告）20 要求进行报告。 1. 实验准备 实验设计（参见 表 1).注意：此行为测试是一项基于气味的Go/Go挖掘任务，其中老鼠必须挖掘以获得高价值或低价值的奖励。它使用一个矩形竞技场（<strong cl…
Representative Results
这里介绍的结果反映了Resasco等人实验19的相关发现。本实验的受试者是18只雌性C57BL / 6NCrl（“C57”）和18只巴尔布/cAnNCrl（“巴尔布”）小鼠。动物在3-4周龄时到达设施，并在混合菌株四重奏25中随机分配到环境丰富或常规住房治疗（分别为EH或CH）。每个笼子里含有一个C57和一个巴尔布，此外还有两个DBA / 2NCrl小鼠在另一个实验中使用。在这里，雌性小鼠的使用?…
Discussion
这里概述的基于气味的挖掘方案和结果证明了这种新型JB任务检测小鼠情感状态变化的能力。因此，这项任务为不同的研究领域提供了一个有价值的工具。与任何JB任务类似，要评估动物影响，动物首先要学会区分线索（步骤4.7.3），并且将模糊刺激解释为中间刺激（步骤5.3）至关重要。虽然很简单，但满足这些要求可能具有挑战性，特别是在实验室小鼠中，过去超过15次开发和实施小鼠JB任务的尝…
Divulgations
The authors have nothing to disclose.
Acknowledgements
作者感谢米格尔·阿亚拉、林赛·基奇纳姆、米歇尔·爱德华兹博士、西尔维娅·林和斯蒂芬妮·德贾尔丁对Reseasco等人19 验证工作的贡献，这些工作是该协议所基于的。我们还要感谢老鼠和我们出色的动物护理技术人员，迈克尔·兰德尔和米歇尔·切普拉克。
Materials
| Absolute ethanol | Commercial alcohol | P016EAAN | Dilute to 70% with distilled water, for cleaning |
| Centrifuge tubes | Fischer | 55395 | 15 mL tubes used to dilute essences here. However, size may be selected based on total volume required for sample size |
| Cheerios (original) | Cheerios | N/A | Commercially available. Used as reward to train animals to enter annex cage for handling |
| Corncob bedding | Envigo | 7092 | Teklad 1/8 corncob bedding used in digging pots and animal cages |
| Cotton pads | Equate | N/A | Commercially available. Modified in lab to fit within tissue cassettes for scent dispensing |
| Digging pots | Rubbermaid | N/A | Commercially available. Containers were modified to fit the apparatus in the lab |
| Dried, sweetened banana chips | Stock and Barrel | N/A | Commercially available. High value reward in JB task |
| JB apparatus | N/A | The apparatus was made in the lab | |
| JWatcher event recording software | Animal Behavior Laboratory, Macquarie University | Version 1.0 | Freely available for download online. Used to score digging behavior during JB task |
| Mint extract | Fleibor | N/A | Commercially "Menta (Solución)". Discriminative stimulus odor |
| Rodent Diet | Envigo | 2914 | Teklad global 14% protein rodent maintenance diets. Low value reward in JB task and regular diet in home cage |
| SAS statistical software | SAS | Version 9.4 | Other comparable software programs (e.g. R) are also appropriate |
| Vanilla extract | Fleibor | Commercially available "Vainilla (Solución)". Discriminative stimulus odor | |
| Video camera | Sony | DCR-SX22 | Sony handycam. |
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