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.