4.3. Nissl and Golgi Staining

For Nissl staining, sections of mouse brains were incubated in 0.1% cresyl violet solution for 5–10 min, then quickly washed in distilled water and dehydrated in increasing ethanol concentrations (70%, 80%, 90%, and 100% ethanol for 2–3 min step by step). The sections were cleared with xylene for 2–5 min, mounted with DPX Mountant (Sigma-Aldrich, St Louis, MO, USA; 44581) for histology and dried in fume hood. The cortical thickness was measured in Nissl-stained sections using the ImageJ software. Golgi staining was performed using the FD Rapid GolgiStain Kit (FD NeuroTechnologies, Columbia, MD, USA) as described previously [49]. For the neuronal morphometric analysis, pyramidal neurons were randomly selected from somatosensory cortical layer II/III. Images were acquired using a BZX microscope at 10× or 40× magnification, and the total dendritic length and Sholl analysis were calculated by Neuron J (ImageJ v1.8.0, NIH).