Fig 2: CRMP2 phosphorylation is enhanced at the presynaptic sites of the dorsal horn. (A) Immunoblots showing the integrity of the synaptic fractionation from lumbar dorsal horn of the spinal cord. The non-PSD fraction was enriched in the pre-synaptic marker Synaptophysin and the PSD fraction was enriched in the post-synaptic marker PSD95. The membrane-associated protein flotillin was used as a loading control. (B) Immunoblots showing the pre-synaptic CRMP2 expression, CRMP2 p32 and CRMP2 p522 levels in the lumbar dorsal horn of the spinal cord of animals having received a spared nerve injury. Synaptophysin shows the consistency of the pre-synaptic material analyzed. Flotilin is used as a loading control. (C) Bar graph showing decreased CRMP2 p32 and increased CRMP2 p522 levels at the pre-synaptic sites in the ipsilateral side of lumbar dorsal horn of the spinal cord. Mean ± s.e.m., *p < 0.05, Mann-Whitney compared to the contralateral side. PSD: post-synaptic density.

Fig 3: Atase1-/- and Atase2-/- improve the APP/PS1 proteotoxicity phenotype.a Combined (both male and female) and sex-specific survival analysis. Data represented are as follows: APP/PS1: n = 112, 41 males, 71 females; APP/PS1; Atase1-/-: n = 74, 37 males, 37 females; APP/PS1; Atase2-/-: n = 52, 27 males, 25 females. *p < 0.05, ***p < 0.0005 via log-rank test. b Thioflavin-S staining of 5 µm paraffin-embedded brain slices. Data are mean ± SD with each data point being the average of technical replicates (a single brain slice) for a given animal, n = 3 animals per genotype. *p < 0.05, **p < 0.005, ***p < 0.0005 via ordinary two-way ANOVA (genotype vs. animal) with Dunnett’s multiple comparison test. c Immunostaining of 5 µm paraffin-embedded brain slices for astrocytes (Gfap) and microglia (Iba1). d Western blotting for Gfap and Iba1 with representative images shown. Data are normalized to ß-actin and represented as the fold change over WT, mean ± SD, n = 3–6 mice per genotype. *p < 0.05 via ordinary one-way ANOVA with Tukey’s multiple comparison test. e Immunostaining of 5 µm paraffin-embedded brain slices for the presynaptic marker synaptophysin and postsynaptic marker Psd-95. Data are mean ± SD with each data point being the average of technical replicates (a single brain slice) for a given animal, n = 3 animals per genotype. **p < 0.005, ***p < 0.0005 via ordinary two-way ANOVA (genotype vs. animal) with Tukey’s multiple comparison test. All analysis was conducted on male mice at 10 months of age.

Fig 4: Methamphetamine self-administration increases the expression of PSD-95 in the dorsal striatum. (a) Schematic of a coronal slice representing the area of tissue punches collected for Western blotting analysis. (b) Representative immunoblots of the various proteins used for Western blotting analysis from control, methamphetamine, and SCH23390 + methamphetamine rats. Corresponding Coomassie staining (Coom) of the membrane is shown as loading control. (c) Quantitative analysis of protein expression in tissue from methamphetamine-exposed rats represented as a percentage change from saline controls. n = 5 saline self-administering rats without SCH23390/i.c. infusions, n = 4 saline self-administering rats with SCH23390, n = 9 methamphetamine self-administering rats without SCH23390/i.c. infusions, and n = 7 methamphetamine self-administering rats with SCH23390 infusion. Data are expressed as mean ± SEM. * p < 0.05 vs. controls; # p < 0.05 vs. methamphetamine rats by one-way ANOVA followed by post hoc analysis.

Fig 5: Knockdown of rat astrocytic Grin2a aggravates Aß-induced synaptotoxicity. (a–e) Electron microscopy results showed the effects of astrocytic Grin2a knockdown and quantification. n = 9–12 synapses from 3 rats/group. AZD, active zone distance; NND, nearest neighbor distance. Bar, 500 nm. (f–h) Representative confocal images (f) and western blot band (h) of PSD95 in the rat hippocampus and quantification (g, h). n = 11–16 sections from 3rats/group. Bar, 200 µm. (i–k) Representative confocal images (i) and western blot band (k) of synaptophysin and quantification (j, k). n = 8 sections from 3 rats/group. Bar, 200 µm. Numerical data are shown as means ± SEM. * vs. vehicle, *p < 0.05, **p < 0.01, ***p < 0.001, # vs. Aß, # p < 0.05, ## p < 0.01, ### p < 0.001