跟踪恐惧条件小鼠
Summary
在接下来的实验中，我们描述了一个协议，用于在小鼠体内微量恐惧条件。这种类型的联想记忆，包括分隔中性刺激和非条件刺激一丝时期。
Abstract
在这个实验中，我们提出了一个方法来衡量学习和记忆。在这里介绍的一丝恐惧条件的协议，存在一个中性刺激和无条件刺激之间的五个配对。有分隔各个空调审判20秒跟踪期。翌日冻结呈现条件刺激（CS）和跟踪一段期间测量。在第三天有一个8分钟的试验来测量情景记忆。代表性的结果是从出席与厌恶无条件刺激（电击），相对于所接收的音调演示，而不无条件刺激小鼠的小鼠。一丝恐惧条件已被成功地用于检测细微的学习和记忆障碍和增强功能未发现与其他恐惧条件反射的方法小鼠。这种类型的恐惧条件的被认为是依赖于内侧前额叶皮质和海马之间的连接。一个电流争论是该方法是否被认为是杏仁核无关。因此，其他的恐惧条件测试是需要检查杏仁核依赖性学习和记忆的效果，如通过延迟恐惧条件反射。
Introduction
在恐惧条件反射的中性刺激（NS）是搭配厌恶无条件刺激（美国）。的NS通常是一个音，并通过反复配对与美国成为条件刺激（CS）。然后将CS可以引起一个条件反应（CR），如冷冻，在没有厌恶美国。常用的恐惧制约的协议是延迟调节。在这个协议中的NS和美国的发病是相连或与在刺激呈现一些重叠。尽管延迟恐惧制约是最常用的类型时空关联空调之一，还有其他一些类型的关联调理时间安排：同步调理，调理落后，并跟踪调节1。在跟踪恐惧条件存在着造成了“跟踪”期间几秒钟的NS和美国之间的经济刺激，无间隔。
一些研究已经报道赤字在痕迹恐惧调理S当神经病变产生的结构，输入到海马区2-5或当药物制剂被用来阻止受体功能的海马。病变海马导致在痕迹条件和上下文调理缺陷，但不影响延迟的恐惧条件8。有几个好处使用痕迹恐惧条件。怕调理协议可以实现在一个为期三天的测试期，并允许海马依赖的记忆，是不是空间相关的。跟踪恐惧条件可以作为一个补充试验的Morris水迷宫，新物体识别测试中，或在调查的海马依赖性记忆的其他迷宫测试。
Protocol
在下面的实验中使用的小鼠产生和安置在贝勒大学在22℃的环境温度下，用14小时光照和10小时黑暗（20:00至下午六小时）昼夜周期。给予小鼠随意获取食物和水。所有程序的小鼠均符合卫生指引全国学院实验动物的护理和使用以及动物方案经贝勒大学动物护理和使用委员会。 概观跟踪空调恐惧任务是基于由Wiltgen及其同事9所述的程序。 <p cl…
Representative Results
对于有代表性的结果，我们提出从较收到的中性刺激，但没有收到非条件刺激（无震动状态）小鼠收到的中性刺激配对与非条件刺激（休克状态）的控制C57BL/6J成年小鼠的数据。它运行这个条件时，首先设置此行为测试，以确定是否该协议已经被正确执行是非常重要的。 图1中的数据代表了一丝恐惧条件测试C57BL/6J小鼠的训练日。它通常没有必要在训练天在所有条…
Discussion
已经有已经阐明了神经电路，跟踪的基础恐惧条件反射的几项研究。追溯恐惧条件，相信涉及的12-14海马CA1区。也有证据表明，内侧前额叶皮质（mPFC的）起着跟踪眼睛一眨不眨空调15大的作用，而内侧前额叶皮质已被发现参与了一丝恐惧条件。一项研究发现，内侧前额叶皮质的神经元在跟踪期间提供持续的活性，从 而提供一个可以追踪间隔17期间维持内存的结构。
…Disclosures
The authors have nothing to disclose.
Acknowledgements
这项工作是由贝勒大学的研究理事会，并授予从癫痫症基金会的研究经费支持。
Materials
| FreezeFrame | Coulbourn | ||
| 30% Isopropanol | Purchase 90% isopropanol and dilute it down to 30% | ||
| 70% Ethanol | |||
| Amp-meter | Med-Associates | ENV-420 | Windows XP, Vista, and 7 Compatible (32-bit only) |
| Digital Sound Level Meter | 33-2055 | ||
| Vanilla Extract | McCormick Pure Vanilla Extract | ||
| Sticky Notes | Post-it | 3X3 inch |
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Cite This Article
Lugo, J. N., Smith, G. D., Holley, A. J. Trace Fear Conditioning in Mice. J. Vis. Exp. (85), e51180, doi:10.3791/51180 (2014).