Journal: Translational Psychiatry

Article Title: Ultramicronized palmitoylethanolamide rescues learning and memory impairments in a triple transgenic mouse model of Alzheimer’s disease by exerting anti-inflammatory and neuroprotective effects

doi: 10.1038/s41398-017-0076-4

Figure Lengend Snippet: Evaluation of neuronal marker expression in hippocampi of 6- and 12-month-old 3×Tg-AD and age-matched Non-Tg mice chronically treated with placebo (open bars) or um-PEA (black bars). ( a , b ) Representative fluorescent photomicrographs of microtubule-associated protein 2 (MAP2) (red) staining in the CA1 region of hippocampi at both 6 and 12 months of age, and ( c ) fluorescence analysis expressed as ΔF/F 0 . Nuclei were stained with DAPI (blue) ( N = 3, in triplicate). ( d ) Representative western blots for MAP2 and ( e ) densitometric analyses normalized with β-actin used as loading controls ( N = 3, in triplicate). The results are expressed as percentage of control (Non-Tg/placebo groups). The data are presented as means ± SEM. Statistical analysis was performed by two-way ANOVA followed by Bonferroni’s multiple-comparison test (* p < 0.05; p ** < 0.01). Scale bar 100 µm

Article Snippet: Overnight incubation at +4 °C was performed with one of the following primary antibodies: rabbit anti-amyloid precursor protein (anti-APP 1:1 000, Cell Signaling, Danvers, MA, USA), rabbit anti-β-secretase (anti-BACE1 1:1 000, Cell Signaling, Danvers, MA, USA), mouse anti-β-amyloid (1:200, Millipore, Darmstad, Germany), rabbit anti-Akt (1:500, Cell Signaling, Danvers, MA, USA), rabbit anti-p[Thr308]Akt (1:5 000, Cell Signalling, Danvers, MA, USA), rabbit anti-glycogen synthase kinase-3β (anti-Gsκ-3β 1:1 000, Cell Signaling, Danvers, MA, USA), rabbit anti-p[Ser9]GSκ-3β (1:1 000, Cell Signaling, Danvers, MA, USA), rabbit anti-p[Ser396]tau (1:1 000, Thermo Fisher Scientific, Waltham, MA, USA), mouse anti-microtubule-associated protein 2 (anti-MAP2 1:250, Novus Biologicals, Littleton, CO, USA), rabbit anti-S100B (1:1 000, Epitomics, Burlingame, CA, USA), rabbit anti-glial fibrillary acidic protein (anti-GFAP 1:25 000, Abcam, Cambridge, UK), rabbit anti-p[Ser536]nuclear factor kappa-light-chain enhancer of activated B cells (anti-p[Ser536]NF-κB p65 1:2 000, Cell Signaling, Danvers, MA, USA), rabbit anti-inducible nitric oxide synthase (anti-iNOS, 1:8 000, Sigma-Aldrich, Milan, Italy), rabbit anti-glutamate transporter GLT-1 (1:1 000, Tocris, Bristol, UK), and mouse anti-glutamine synthetase clone GS-6 (1:1 000, Millipore, Darmstad, Germany).

Techniques: Marker, Expressing, Staining, Fluorescence, Western Blot, Comparison