PKC53e RNAi preliminary experiments

Description of the experiment and the figures <em>Drosophila </em>self-learning. The flies are tethered to a torque meter which measures the angular momentum around the fly's vertical body axis (yaw torque), caused by attempted turning maneuvers. Attempted left (or right) turning maneuvers are punished during training phases and no predictive stimuli are present. Thus, the only predictor of punishment is the behavior of the fly. If the fly continue to avoid the punished behavior when laser is absent during test phases, we talk about self-learning. Scoring and learning protocol. A score corresponding to the proportion of time the fly is doing the safe turning attempt is calculated for each 2 minutes period. Before training the score is around 0 (no preference). During training phases, the score rises since the fly avoid doing the laser punishment. After two rounds of 4 minutes training separated by a 2min test, a final 2 minutes test round is performed. The performance during these two minutes is used for statistical analysis. After the experiment, the torque force of the fly is tested again using its optomotor response and the accuracy of the laser position (the fly is sacrificed by letting the laser on for about 15s). If any of these two tests is not satisfactory, the data is not included in the analysis. <br><br> <strong>Representation legend .</strong> The score during the final test is depicted in boxplots (bar is the median, boxes are quartiles, bars are 100% of the data not considering the outliers presented by points). Detailed information. Genetics: parental lines UAS-PKC53e.RNAi lines (V: from VDRC, T: Trip line) males were crossed to elavGal4; tubgal80ts females. Only females were tested, Flies were grown at 25 degrees and put for two days at 32 degrees after eclosion before being tested.. NB: data where the pretest score was higher than 0.6 were excluded from the analysis. Full genotype of the flies tested is indicated below: elavG4;tG80 x CS : UAS_PKC53eRNAi_vienna/+ . CS x 53eRi_V : elavG4/+;tubGal80ts/+;UAS_PKC53eRNAi_vienna/+ . CS x 53eRi_T : elavG4/+;tubGal80ts/+ . elavG4;tG80 x 53eRi_T : elavG4/+;tubGal80ts/UAS_PKC53eRNAi_Trip . elavG4;tG80 x 53eRi_V : UAS_PKC53eRNAi_Trip/+ . ------ flybase numbers: tubGal80^ts= P{tubP-GAL80ts} (FBtp0017264)[on the 2nd chromosome], elavGal4= FBti0002575 UAS-PKC53e.RNAi_V= FBti0159711 UAS-PKC53e.RNAi_T= FBti0128750

实验与图示说明：<em>Drosophila</em>自学习实验。将果蝇固定于扭矩仪上，该仪器可测量果蝇因尝试转向动作而产生的、绕身体垂直轴的角动量（偏航扭矩，yaw torque）。训练阶段中，果蝇尝试向左（或向右）转向的动作会被施加惩罚，且此时无任何预测性刺激存在。因此，惩罚的唯一预测因子即为果蝇自身的行为。若在测试阶段无激光刺激时，果蝇仍能持续避免曾被惩罚的转向动作，则可称其发生了自学习。

评分与学习范式：每2分钟为一个计时周期，计算对应周期内果蝇做出安全转向动作的时长占比，以此作为评分。训练前，该评分约为0（无偏好性）。训练阶段中，由于果蝇会主动规避会触发激光惩罚的动作，评分会随之升高。在两轮各4分钟的训练（中间间隔2分钟测试环节）结束后，开展一轮最终的2分钟测试。将这2分钟内的表现数据用于后续统计分析。实验结束后，借助果蝇的视动反应（optomotor response）再次测试其扭矩响应，并验证激光定位的准确性；随后将激光持续照射约15秒以处死果蝇。若上述两项测试任一未达标准，则该组数据将被排除出最终分析。

<strong>图示说明</strong>。最终测试阶段的评分以箱线图（boxplot）呈现：箱线中间的横线为中位数，箱体上下边界为四分位数，须线覆盖除去异常值（以散点表示）外的全部数据范围。

详细信息：遗传学背景为，将UAS-PKC53e.RNAi品系（V：来自维也纳果蝇资源中心VDRC，T：Trip文库品系）的雄性成虫与elavGal4; tubgal80ts品系的雌性成虫进行杂交，仅选用杂交后代的雌性果蝇开展实验。果蝇饲养温度为25℃，羽化后转移至32℃环境培养2天再进行测试。注意：预测试评分高于0.6的数据集将被排除出分析。

受试果蝇的完整基因型如下所示：
elavG4;tG80 × CS：UAS_PKC53eRNAi_vienna/+
CS × 53eRi_V：elavG4/+;tubGal80ts/+;UAS_PKC53eRNAi_vienna/+
CS × 53eRi_T：elavG4/+;tubGal80ts/+
elavG4;tG80 × 53eRi_T：elavG4/+;tubGal80ts/UAS_PKC53eRNAi_Trip
elavG4;tG80 × 53eRi_V：UAS_PKC53eRNAi_Trip/+

——果蝇数据库（FlyBase）编号：tubGal80^ts = P{tubP-GAL80ts}（FBtp0017264，位于2号染色体）；elavGal4 = FBti0002575；UAS-PKC53e.RNAi_V = FBti0159711；UAS-PKC53e.RNAi_T = FBti0128750