Shoji Hirotaka, Takao Keizo, Hattori Satoko, Miyakawa Tsuyoshi
Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University; Japan Science and Technology Agency, Core Research for Evolutionary Science and Technology (CREST).
Japan Science and Technology Agency, Core Research for Evolutionary Science and Technology (CREST); Center for Genetic Analysis of Behavior, National Institute for Physiological Sciences, National Institutes of Natural Sciences.
J Vis Exp. 2014 Mar 1(85):50871. doi: 10.3791/50871.
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.
情境性和线索性恐惧条件反射测试是评估小鼠学习和记忆环境线索与厌恶经历之间关联能力的行为测试之一。在该测试中，将小鼠放入条件反射箱，给予条件刺激（听觉线索）和厌恶非条件刺激（足部电击）配对。经过一段延迟时间后，让小鼠暴露于相同的条件反射箱以及呈现听觉线索的不同形状的箱子中。测试期间的僵立行为被测量作为恐惧记忆的指标。为了自动分析行为，我们使用ImageFZ应用软件程序开发了一个视频分析系统，该程序可在http://www.mouse-phenotype.org/免费下载。在此，为了展示我们方案的细节，我们展示使用ImageFZ系统在C57BL/6J小鼠中进行情境性和线索性恐惧条件反射测试的过程。此外，我们通过比较ImageFZ系统或基于光电束的计算机测量系统测量的僵立时间与人类观察者评分，验证了我们的方案和视频分析系统的性能。如我们的代表性结果所示，ImageFZ获得的数据与人类观察者分析的数据相似，表明使用ImageFZ系统进行行为分析具有高度可靠性。本视频文章提供了有关测试程序的详细信息，并将促进对实验情况的理解。
