Confocal image stacks of GFP expression in Drosophila forelegs driven by Gal4 driver expression in foreleg motor neurons

Like the vertebrate spinal cord, the insect ventral nerve cord (VNC) mediates limb sensation and motor control. We applied automated tools for electron microscopy volume alignment, neuron segmentation, and synapse prediction toward creating a connectome of an adult female Drosophila VNC. To interpret a connectome, it is crucial to know its relationship with the rest of the body. We therefore mapped the muscle targets of leg and wing motor neurons in the connectome by comparing their morphology to genetic driver lines, dye fills, and X-ray nano-tomography of the fly leg and wing. The dataset made available here contains the confocal image stacks of the legs of each driver line. We imaged only driver lines for which we found an image of a single motor neuron labeled by MCFO, from the collection on the https://neuronbridge.janelia.org website. Thus, the dendritic morphology of the MNs that are visible in these images stacks can be found by cross referencing the alphanumeric code for the Ga..., Confocal imaging
Fly prothoracic (front) legs were immersed in a 4% formaldehyde (PFA) PBS solution for 20 minutes, followed by three rinses in PBS with 0.2% Triton X-100 (PBT). The legs were then incubated in a PBS solution containing 1:50 phalloidin (Alexa-phalloidin-647, Fisher A222287) and the following reagents that improve tissue penetrance: 1% Triton X-100, 0.5% DMSO, 0.05 mg/ml Escin (Sigma-Aldrich, E1378), and 3% normal goat serum. Legs were incubated for one week at 4 °C with occasional rocking. After staining, legs were rinsed 3x with PBS-Tx, 1 rinse with PBS, and were mounted onto slides in Vectashield. To image MNs innervating the coxal muscles in the thorax, the fly was fixed as above, then hemisected along the parasagital plane with a fine razor blade in Tissue-Tek O.C.T. compound (Sakura 4583) frozen for 10 seconds on dry ice. Hemisected thoraces were rinsed 3x in PBT and stained as above.
Mounted legs or thoraxes were imaged on a Confocal Olympus FV1000. At least one im..., , # Confocal image stacks of GFP expression in Drosophila forelegs driven by Gal4 lines labelling foreleg motor neurons

[https://doi.org/10.5061/dryad.k0p2ngfg0](https://doi.org/10.5061/dryad.k0p2ngfg0)

Like the vertebrate spinal cord, the insect ventral nerve cord (VNC) mediates limb sensation and motor control. We applied automated tools for electron microscopy volume alignment, neuron segmentation, and synapse prediction toward creating a connectome of an adult female *Drosophila* VNC. To interpret a connectome, it is crucial to know its relationship with the rest of the body. We therefore mapped the muscle targets of leg and wing motor neurons in the connectome by comparing their morphology to genetic driver lines, dye fills, and X-ray nano-tomography of the fly leg and wing. The dataset made available here contains the confocal image stacks of the legs of each driver line. 

## Description of the data and file structure

Meissner et al. ([https://doi.org/10.7554/eLife.80660](https:...

与脊椎动物脊髓类似，昆虫腹神经索（ventral nerve cord, VNC）介导肢体感知与运动调控。本研究借助自动化工具完成电子显微镜体数据对齐、神经元分割与突触预测，旨在构建成年雌性果蝇腹神经索的连接组。要解析连接组，明确其与机体其余部分的关联至关重要。为此，本研究通过将连接组内腿部与翼部运动神经元的形态与遗传驱动系、染料填充结果以及果蝇肢体的X射线纳米断层扫描数据进行比对，绘制了这些运动神经元的肌肉靶标图谱。本数据集包含各遗传驱动系腿部的共聚焦图像栈。本研究仅对https://neuronbridge.janelia.org网站收录的、经MCFO标记的单个运动神经元对应的遗传驱动系进行成像。因此，通过交叉引用Ga...的字母数字编码，即可获知这些图像栈中可见的运动神经元树突形态。
### 共聚焦成像
果蝇前胸（前）腿置于4%甲醛（多聚甲醛，PFA）的磷酸盐缓冲液（PBS）中固定20分钟，随后用含0.2% Triton X-100的PBS（PBT）漂洗三次。随后将腿部置于含1:50浓度鬼笔环肽（Alexa-鬼笔环肽-647，Fisher A222287）以及以下增强组织渗透性试剂的PBS溶液中孵育：1% Triton X-100、0.5%二甲基亚砜（DMSO）、0.05 mg/ml七叶皂苷（Sigma-Aldrich, E1378）与3%正常山羊血清。腿部于4℃下孵育一周，期间定期摇动。染色完成后，腿部先用PBS-Tx漂洗三次，再用PBS漂洗一次，随后用Vectashield封片于载玻片上。若要成像支配胸部基节肌肉的运动神经元，则按上述方法固定果蝇，随后用细剃须刀沿矢状旁平面对半剖开，将其置于干冰上冷冻10秒的Tissue-Tek O.C.T包埋剂（Sakura 4583）中。对半剖开的胸部经PBT漂洗三次后，按上述方法进行染色。
封片后的腿部或胸部在奥林巴斯FV1000共聚焦显微镜上成像。至少完成一次成像……
# 标注前腿运动神经元的Gal4驱动系介导的果蝇前腿GFP表达共聚焦图像栈
[https://doi.org/10.5061/dryad.k0p2ngfg0](https://doi.org/10.5061/dryad.k0p2ngfg0)

与脊椎动物脊髓类似，昆虫腹神经索（ventral nerve cord, VNC）介导肢体感知与运动调控。本研究借助自动化工具完成电子显微镜体数据对齐、神经元分割与突触预测，旨在构建成年雌性果蝇腹神经索的连接组。要解析连接组，明确其与机体其余部分的关联至关重要。为此，本研究通过将连接组内腿部与翼部运动神经元的形态与遗传驱动系、染料填充结果以及果蝇肢体的X射线纳米断层扫描数据进行比对，绘制了这些运动神经元的肌肉靶标图谱。本数据集包含各遗传驱动系腿部的共聚焦图像栈。

## 数据与文件结构说明
Meissner等人（[https://doi.org/10.7554/eLife.80660](https:...）