Amx1200

PFA-fixed slices from treated mice brains were stained in cresyl violet solution (0.2% cresyl violet (Sigma), 0.5% glacial acetic acid, 0.01 M sodium acetate) for 20 min at 37 °C. Sections were then sequentially dehydrated with rapid washes in: 70% ethanol, 95% ethanol, 100% ethanol, methanol, 1:1 methanol-xylene. Finally, glasses were covered by using Eukitt (Sigma, Saint Louis, USA). Pictures of Nissl-stained brain sections were acquired with a Nikon AMX1200 digital camera on a Nikon E800 Microscope (× 4 magnification). Bregma position was assigned to each slice following [38 ] and then the FIJI software was used to measure cortical thickness [31 (link)]. First, we produced a densitometrical plot profile from a line spanning the cortex perpendicularly from the pial surface to the white matter underlying cortical layer VI. We then identified transitions among layers as variations in the densitometrical plot profile, which reflect cell density changes between cortical layers, and measured their lengths in the plot (see Supplementary Fig. S3). This approach allowed us to measure the total cortical thickness in S1 and M1, and the relative thickness of cortical layers I, II/III–IV, and V–VI in the S1. Eight mice per group and 10–15 slices per mice were analyzed.

Thickness of the total cortex and of its layer of WT and KO mice untreated or treated with antidepressants were analyzed on 3 adjacent sections, taken from the beginning of the barrel cortex to 1200 μm. Pictures were taken using a high-definition Nikon AMX1200 digital camera on a Nikon E800 Microscope (10X magnification). Gray level profile along a line (width = 300) spanning perpendicularly the barrel cortex or the hippocampus was determined by ImageJ software.