Immunohistochemical Analysis of Synaptic Proteins

This procedure was performed by a standard indirect method as described previously [28 (link)]. Primary antibodies and dilutions were as follows: anti-COX IV (1:250; # ab16056, Abcam), anti-DRP1 (1:250; # ab56788, Abcam), anti-Mfn2 (1:250; # ab50838, Abcam), anti-proBDNF (1:250; # ab72440, Abcam), anti-mBDNF (1:250; # GF35L, Millipore), anti-TrkB (1:250; # ab18987, Abcam), anti-phospho-TrkB (phospho TrkB Y817; 1:250; # ab81288, Abcam), anti-p75^NTR (1:250; # ab93934, Abcam), anti-synapsin I (1:250; # ab64581, Abcam), anti-PSD-95 (1:250; # ab12093, Abcam), and anti-amyloid beta peptide (MOAB-2; 1:250; # MABN254, Millipore). After incubation with the respective secondary antibodies DyLight-650 or Alexa Fluor 488 (## ab96886, ab150073, Abcam) diluted 1:250, the slices were coverslipped with the Fluoro-shield mounting medium containing 4′,6-diamidino-2-phenylindole (DAPI; # ab104139, Abcam) and examined under an Axioplan 2 microscope (Zeiss).