Imaging data and 3D reconstruction of neuronal dendrites

Three pairs of WT and Zfp661 KO mouse littermates, including both male and female pairs aged between 60 to 66 days, were used in this assay. The brains were isolated and processed for Golgi-Cox staining using FD Rapid GolgiStain Kit (FD NeuroTechnologies, PK401) according to the manufacturer’s protocol. The pyramidal neurons in the somatomotor area of Layer 2/3 (Bregma 1.70 mm to 1.18 mm) were sampled and imaged using a Leica DM6000B (20X/0.70) microscope and LAS X software with optimal z-step of 0.52 μm and the same settings for all imaging. To better trace neuronal dendrites from Golgi-Cox staining, Z-stacked images were pre-processed using Fiji/ImageJ2 (v2.9.0/1.53t) as follows: the image format was transformed into 8-bit type, both the signals and LUT were inverted, and the scale unit was set to microns. All dendrites from sampled neurons were traced using the SNT toolbox in Fiji/ImageJ2 with the “A* search algorithm” enabled, and the trace for each dendrite was further manually verified. The uploaded zip file contains the mouse information, preprocessed images and neuronal dendrite traces.本实验采用3对野生型（WT）与Zfp661基因敲除（Zfp661 KO）的同窝小鼠，涵盖雌雄个体，日龄为60至66日。分离小鼠脑组织后，依照制造商操作规范，使用FD Rapid GolgiStain染色试剂盒（FD NeuroTechnologies，货号PK401）完成高尔基-考克斯（Golgi-Cox）染色。选取Bregma坐标1.70 mm至1.18 mm范围内的第2/3层躯体运动区锥体细胞，通过徕卡DM6000B（20X/0.70）显微镜及LAS X软件完成成像，设置最优z轴步距为0.52 μm，所有成像环节均保持参数一致。为更精准地示踪高尔基-考克斯染色后的神经元树突，使用Fiji/ImageJ2（版本v2.9.0/1.53t）对Z轴堆叠图像进行预处理，具体步骤如下：将图像格式转换为8位灰度图，反转图像信号与查找表（Lookup Table, LUT），并将标尺单位设置为微米。使用Fiji/ImageJ2内置的SNT工具箱，启用A*搜索算法对所有取样神经元的树突进行追踪，并对每条树突的追踪结果进行人工复核验证。本次上传的压缩包包含小鼠样本信息、预处理后的图像以及神经元树突追踪数据。