Quantifying Brain-Derived Neurotrophic Factor

For brain function, brain slices were stained for brain-derived neurotrophic factor (BDNF), as described previously (Hovens et al., 2014b (link)). In brief, sections were blocked for 1 h with 5% normal goat serum, then incubated with 1:1,000 rabbit-anti BDNF antibody (Alomone Labs, Israel) in 1% BSA, followed by incubation with 1:5,000 goat-antirabbit secondary antibody (Jackson, Wet Grove, USA). Photographs were taken at 50× magnification (Toth et al., 2022b (link)) from the different areas of the dorsal hippocampus, CA1, CA3, DG, and hilus, and BDNF expression was obtained as corrected optical density (Image-J) compared to an underlying reference area (Hovens et al., 2014b (link)).