Wild Type DCZ All Time Points Contextual Fear Conditioning Data

Contextual Fear Conditioning: Mice underwent a 3-day contextual fear conditioning protocolduring their light cycle (6am-6pm). Mice were habituated for 3 days prior to conditioning day (Day 0) for 1 hour in squads of 4 to a neutral habituation room. To create distinct and similar contextual environments, chambers and interchangeable inserts from MedAssociates were used in two rooms. Contexts A and B were presented in the same room, and context C was presented in a different room. Each context was assigned a specific set of features (floor, roof, scent, sound, light, and transport vessels) as described in Extended data Figure 3-1. Context B is meant to closely resemble A, thus only 2 of 6 features (floor and transport) were altered (Extended data Figure 3-1). Mice were injected intraperitoneally (i.p.) with saline or 1.0 mg/kg DCZ (Ferrari, Ogbeide-Latario et al. 2022) 10 minutes prior to being placed into behavior boxes (MedAssociates) on all days or only on a specific day as described in the results section. During acquisition, mice received 4 shocks (1 mA, 2-second duration, 1-minute inter-shock interval), delivered after an initial 3-minute baseline period (Zelikowsky, Bissiere et al. 2013). One minute after the final shock, acquisition ended, and mice were transported back to their home cage. 24 hours later, on test day 1, mice were habituated and injected as described earlier. Conditioned available under aCC-BY-NC-ND 4.0 International license. Mice were exposed to context A for 8 minutes or to the “similar” context B (counter balanced). Mice were returned to their home cage in the vivarium for a minimum of 5 hours before being tested in the alternative context. Twenty-four hours later, this process was repeated for test day 2, with the distinct, “neutral” context C replacing the “similar” context B. To analyze behavior, component files were made to extract the desired data using automated near-infrared video tracking equipment and computer software (VideoFreeze, MedAssociates). For conditioning day 0, the 3-minutes prior to the first shock were extracted to analyze baseline percent component freezing and motion index. The 30 seconds preceding each shock were also extracted to analyze the percent component freezing for fear acquisition curves. For test days 1 and 2, the percent time freezing was calculated for the entire 8-minute session. All mice used for behavior were verified for correct virus expression and targeting. Mice were excluded from all analyses if they did not express virus in the CA3/hilus region or had more infection outside the CA3/hilus region than inside (Extended Data Figure 3-1).

情境恐惧条件反射（Contextual Fear Conditioning）：小鼠在光照周期（上午6时至下午6时）内接受为期3天的情境恐惧条件反射实验方案。在条件反射实验当天（第0天）前，小鼠需连续3天以4只为一组，在中性适应笼室中适应1小时。为构建差异化与相似性兼具的情境环境，实验在两间房间内使用了MedAssociates公司的行为箱与可更换衬垫。情境A与B设置于同一房间，情境C则设于另一独立房间。如补充数据图3-1所述，每个情境均配备一套专属特征，包括地板、顶板、气味、声音、光照与转运容器。情境B旨在高度模拟情境A，因此仅调整了6项特征中的2项（地板与转运容器），详见补充数据图3-1。小鼠需在放入MedAssociates公司行为箱前10分钟，接受腹腔注射（intraperitoneally，i.p.）生理盐水或1.0 mg/kg的DCZ（Ferrari、Ogbeide-Latario等，2022），注射时机为所有实验日或仅结果部分所述的特定实验日。在习得阶段，小鼠先经历3分钟的基线期，随后接受4次足底电击（电流1 mA，时长2秒，电击间隔1分钟），该流程参考Zelikowsky、Bissiere等（2013）的方法。末次电击结束1分钟后，习得阶段结束，小鼠被送回饲养笼。24小时后，即测试日1，小鼠按照前述流程进行适应与注射。本数据集采用CC-BY-NC-ND 4.0国际许可协议进行授权。小鼠被暴露于情境A中8分钟，或暴露于“相似”情境B中，实验采用平衡抵消设计。小鼠被送回饲养室的饲养笼中，至少静置5小时后，再接受另一情境的测试。24小时后，该流程于测试日2重复进行，此时将“相似”情境B替换为差异化的“中性”情境C。行为学分析环节中，研究人员借助自动化近红外视频追踪设备与计算机软件（VideoFreeze，MedAssociates公司）生成组分文件，提取目标数据。针对第0天的条件反射实验，研究人员提取首次电击前3分钟的行为数据，用于分析基线冻结行为占比与运动指数；同时提取每次电击前30秒的行为数据，用于分析恐惧习得曲线的冻结行为占比。对于测试日1与测试日2，则计算整个8分钟实验时段内的冻结时间占比。所有参与行为学实验的小鼠均经过验证，确认其病毒表达与靶向定位均符合要求。若小鼠未在CA3/门区（CA3/hilus region）表达病毒，或病毒感染在CA3/门区外的范围大于区内，则将其排除于所有分析之外，详见补充数据图3-1。