To confirm that a diminished apoptotic rate was responsible for the survival of BrdU-positive cells in bim−/− and puma−/− mice, brain sections from mice of all genotypes were terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelled (TUNEL) for apoptotic DNA strand breaks.
To determine whether the severity of changes in behavior observed in Tg20 and Prnp −/− after KA treatment correlated to neuron death in the hippocampus, we carried out Fluoro Jade-B (FJ-B) histochemical staining in coronal brain sections from KA-treated mice from all three experimental groups (Fig. 1).
In addition, in brain sections from APP23 mice of all ages examined in the present study and regardless of the Abca1 genotype, the extracellular staining with A11 antibody was spatially segregated from the plaques, similar to human AD brain (Kayed et al., 2003).
In all brain sections from transgenic APP23, Tg2576 and APPswe-PS1dE9 mice incubated with [ 18 F]flutemetamol in vitro, the bound radioactivity co-localized well with fibrillar Aβ plaques stained with ThS and anti-Aβ1-40 antibody (Figure 4).
All brain sections from different animal groups were simultaneously run to ensure identical staining conditions.
Hematoxylin and eosin (H&E) staining was performed on at least two brain sections from each mouse to align all brains to approximately 1.3 mm lateral to the midline using a mouse brain atlas [ 52].
For all brain sections from the 18 h recovery time point, except for the pituitary gland due to insufficiency of available material, protein was extracted according to assay kit recommendations (Assay Designs, Ann Arbor, USA).
To determine whether EB101 vaccine attenuates and/or reverses the massive development of β-amyloid plaques, brain sections from wild-type and transgenic mice of all experimental groups were immunostained with the specific antibody recognizing A β1 42 epitopes.
Histology and immunohistochemistry were carried out as previously described [23] on brain sections from frontal, temporal and parietal neocortices (the occipital cortex was unavailable), neo-striatum, thalamus, cerebellar hemisphere and on sections from all received tissues.
