A reversal in sensory processing accompanies ongoing ecological divergence and speciation in Rhagoletis pomonella

Uploaded as part of this dataset are raw files collected in pursuit of results part b (antennal lobe input), part c (activity in the antennal lobe), and part d (antennal lobe output) of our manuscript. All three of these types of files include both apple and hawthorn race flies, as we were interested in how they diverged at these different levels of olfactory processing, and are labeled by host race and, usually, odor stimulus given to cause response captured in the particular file. The data for part b, antennal lobe input, consists of 19 ".oib" files, labeled with "partB", then numerically as "OSN_1" through "OSN_19", according to supplementary table s1 in the manuscript, and with whether they came from an "Apple" or "Hawthorn" race fly. They are the raw stacks of images obtained via confocal microscopy, consisting of one channel capturing the neuropil marker at the 647 wavelength and one channel capturing the neurobiotin tracer backfilling an OSN that responded to 3MB and BH at the 488 wavelength. Magnification was 40x, using an oil objective. Files can by opened in FIJI (Fiji: an open-source platform for biological-image analysis. Nat. Methods 9, 676–82). These files are raw files, before all processing. To replicate the results shown in the manuscript in Figure 3 and Supplementary Figure S2, one should consult the methods and supplementary methods of the manuscript, or the uploaded ReadMe file. The data for part c, activity in the antennal lobe, consists of 26 ".tif" files, labeled "partC", then the numeric file name, followed by "Apple" or "Hawthorn" and then further followed by the odor stimulus during that particular trace "BH" or "3MB". Focused on the antennal lobe after it had been incubated with Calcium Green AM-ester, these traces show 10 seconds, recorded at 4 Hz on a CCD camera with a 20x water-immersion objective, wherein the given odor stimulus (BH or 3MB) was introduced at the 2 second time point. A peak of activity shown by the calcium-sensitive fluorescent dye can be seen for each of these traces within 2.5 seconds after the 2 second introduction of odor. Again, these are raw data files. To replicate results, corrections for movement and bleaching were undertaken, again using FIJI, as well as multi-step manual landmark registration, to produce Figure 4 and Supplementary Figures S3 and S4. For details on processing and analysis, one should consult the methods and supplementary methods of the manuscript or the uploaded ReadMe file. The data for part d, antennal lobe output, consists of 10 ".oib" or ".oir" files, labeled with "partD", then the numerical file name, followed by "Apple" or "Hawthorn", then further described by the odor stimulus that the filled antennal lobe neuron responded to, "BH", or "3MB" and the glomerulus/i filled in that case, either "DM1", "VP1", or "VP2" or some combination thereof. This data for all 10 of these images is summarized by Supplementary Figures S5 and S6, with detailed analyses shown in Figure 5. Uploaded here are the raw data files, taken via confocal microscopy, showing one channel capturing the neuropil marker at the 647 wavelength and one channel capturing the neurobiotin tracer filling the ALN at the 488 wavelength. Magnification was 40x, using an oil objective. For details on the processing and analysis of these raw files, one should consult the methods and supplementary methods in the manuscript or the uploaded ReadMe file.

本数据集上传的原始文件，采集自本论文中第B部分（触角叶输入）、第C部分（触角叶活动）及第D部分（触角叶输出）的相关研究，以获取对应实验结果。三类文件均包含苹果寄主种群与山楂寄主种群的果蝇样本——研究团队旨在探究二者在嗅觉加工的不同层级上的分化模式，所有文件均按寄主种群进行标注，且通常会标注用于诱发该文件中记录的神经反应的气味刺激物。第B部分（触角叶输入）的数据包含19个.oib格式文件，命名格式为"partB"后接编号"OSN_1"至"OSN_19"（对应论文补充表S1），并标注样本来源为"Apple"（苹果种群）或"Hawthorn"（山楂种群）果蝇。此类文件为共聚焦显微镜采集的原始图像堆栈，包含两个成像通道：一个通道以647波长捕获神经纤维丛标记物信号，另一个通道以488波长捕获对3MB与BH气味产生反应的嗅觉感觉神经元（Olfactory Sensory Neuron，OSN）的神经生物素示踪信号。实验采用油浸物镜，放大倍数为40倍。此类文件可通过FIJI（Fiji：开源生物图像分析平台，《自然-方法》9卷, 676–82页）打开，均为未经任何预处理的原始文件。若需复现论文中图3及补充图S2的实验结果，请参考论文的方法与补充方法部分，或本数据集附带的ReadMe文件。第C部分（触角叶活动）的数据包含26个.tif格式文件，命名格式为"partC"后接数字文件名，随后标注"Apple"或"Hawthorn"，最后标注该次记录中使用的气味刺激物"BH"或"3MB"。此类数据针对经钙绿AM-酯（Calcium Green AM-ester）孵育后的触角叶，通过CCD相机以4 Hz帧率记录了10秒的活动信号，实验采用20倍水浸物镜，气味刺激物（BH或3MB）于记录开始后的第2秒时施加。每段记录均可在气味施加后的2.5秒内观察到钙敏感荧光染料所反映的活动峰值。此类文件同样为原始数据文件。若需复现论文中图4及补充图S3、S4的实验结果，需先通过FIJI进行运动校正与光漂白校正，并结合多步骤人工地标配准完成数据处理。有关处理与分析的详细细节，请参考论文的方法与补充方法部分，或本数据集附带的ReadMe文件。第D部分（触角叶输出）的数据包含10个.oib或.oir格式文件，命名格式为"partD"后接数字文件名，随后标注"Apple"或"Hawthorn"，并进一步标注被标记的触角叶神经元（Antennal Lobe Neuron, ALN）所响应的气味刺激物"BH"或"3MB"，以及此时被填充的嗅小球，包括"DM1"、"VP1"、"VP2"或其组合。上述10幅图像的相关数据已在补充图S5、S6中进行汇总，详细分析内容见论文图5。本数据集上传的此类原始文件通过共聚焦显微镜采集，包含两个成像通道：一个通道以647波长捕获神经纤维丛标记物信号，另一个通道以488波长捕获填充触角叶神经元的神经生物素示踪信号。实验采用油浸物镜，放大倍数为40倍。有关此类原始文件的处理与分析细节，请参考论文的方法与补充方法部分，或本数据集附带的ReadMe文件。