Journal: Molecular Neurodegeneration

Article Title: Trem2 deficiency differentially affects phenotype and transcriptome of human APOE3 and APOE4 mice

doi: 10.1186/s13024-020-00394-4

Figure Lengend Snippet: Trem2 deficiency impacts cognition, plaque compaction, and microglia recruitment in APP/E3 and APP/E4 mice. a Schematic timeline showing groups and experimental procedures of 6.5-month-old mice used for behavioral, histological, and transcriptional analysis (Figs. 1, , , and ). b Novel object recognition (NOR), and Contextual fear conditioning. Analysis by two-way ANOVA showed no interaction between APOE isoform and Trem2 status and a significant main effect of APOE isoform (F (1, 49) = 13.28, p < 0.01) and Trem2 status (F (1, 49) = 11.06, p < 0.01) for NOR ( a , b ), and Contextual fear conditioning ( c, d ) APOE isoform effect (F (1, 50) = 11.39, p < 0.01) and Trem2 status (F (1,50) = 10.86, p < 0.01). ** p < 0.01; * p < 0.05, Sidak multiple comparisons test. n = 6–14 mice per group. For APP mice n = 6–7 mice/genotype/sex (12–14 mice/genotype). For non-APP mice, n = 4–7 mice/genotype/sex (8–14 mice/genotype). On the graphs, red symbols indicate female and black symbols indicate male mice. c Representative images of X34 and OC labeled amyloid deposits showing core-bound and non-core bound OC. d Bar plot depicting the ratio of non-core bound OC to total OC. n = 15–26 mice per group. e Representative images of glial cells (Iba1+ microglia and GFAP+ astrocytes) recruited to amyloid plaques. f Bar plots depicting the number of microglia nuclei within 60 μm of plaque border. g Bar plots depicting the number of astrocyte nuclei within 60 μm of plaque border. n = 80–120 plaques from 6 mice per group. h Representative images of X34 and LAMP1 label showing neuronal dystrophy surrounding amyloid deposits. X34 is shown as a blue region of interest defined by NIS elements thresholding. i Bar plot depicting the area of plaque-associated LAMP1 staining. Analysis by two-way ANOVA showed no interaction between APOE isoform and Trem2 status and a significant main effect of APOE isoform (F (1, 476) = 25.41, p < 0.0001) and Trem2 status (F (1, 476) = 4.99, p < 0.05) for LAMP1 area. Sidak multiple comparison test found no difference in plaque-associated LAMP1 staining area between APP/E3 vs APP/E3/Trem2 ko or APP/E4 vs APP/E4/Trem2 ko . n = 120 plaques from 4 mice per group. j Representative images of plaque-associated APOE (green) and TR (red) staining to visualize compact amyloid plaques. k Bar plots showing the area of APOE staining that surrounds TR positive amyloid plaques. n = 874–2719 plaques from 4 to 6 mice per group. l Bar plots depicting Apoe gene expression as identified by RNA-seq, which closely follows the pattern of plaque-associated APOE protein levels. For histological analyses, one-way ANOVA was used followed by Tukey’s multiple comparison test. m Representative images of FISH analyses of gene expression near amyloid plaques ( Tmem119 – green, Apoe – Pink, Nuclei – Blue). n Bar plot depicting the Apoe gene expression within Tmem119 -positive microglia cells. The intensity of Apoe FISH signal was normalized to the number of Tmem119 -positive microglial cells. n = 279–313 microglia per group. Bars represent mean ± SEM, with all red bars = APP/E3, orange = APP/E3/Trem2 ko , purple = APP/E4, and blue = APP/E4/Trem2 ko . *** p < 0.001; ** p < 0.01; * p < 0.05; NS = No Signifincace

Article Snippet: On Day 1, training phase, mice were placed in a conditioning chamber (Stoelting Co., USA) for 5.5 min.

Techniques: Labeling, Staining, Comparison, Gene Expression, RNA Sequencing

Journal: Translational Psychiatry

Article Title: TOB is an effector of the hippocampus-mediated acute stress response

doi: 10.1038/s41398-022-02078-7

Figure Lengend Snippet: Behavioral analyses in Tob -WT and KO mice and after overexpression of mouse TOB using AAV (hSyn-mTob) A – D . A Contextual fear conditioning and extinction expressed as percentage of time spent freezing. Two-way ANOVA followed by Bonferoni’s post-hoc test for multiple comparisons. B The forced swim test presented as a percentage of immobile time. One-way ANOVA followed by Bonferoni’s post-hoc test for multiple comparisons. C Elevated-plus maze showing the percentage of time spent in open arm. One-way ANOVA followed by Bonferoni’s post-hoc test for multiple comparisons. D Open field test showing the percentage of time spent in center region. One-way ANOVA followed by Bonferoni’s post-hoc test for multiple comparisons. Behavioral analyses in hippocampal-specific Tob -KO mice (E-I). E Schematic diagram showing the method for generation of hippocampal-specific Tob -KO (hsTobKO) mice through injection of adeno-associated virus expressing Cre recombinase under the hSyn promoter (AAV_hSyn_Cre) in mice having LoxP sequences flanking both sides of the Tob gene ( Tob fl/fl ). F Contextual fear conditioning and extinction in hsTobKO presented as percentage of time showing freezing. Two-way ANOVA followed by Bonferoni’s post-hoc test for multiple comparisons. G The forced swim test is presented as percentage of time spent immobile. H The elevated-plus maze showed as the time spent in the open arm. I Open field test showing the percentage of time spent in the center region. Unpaired t-test. All values represent means ± SEMs. ns non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Briefly, on training day, mice were placed in a conditioning chamber (CL-3002L, O’Hara & Co Ltd., Japan) for 2 min to habituate, and then presented with a conditioning stimulus (CS) of a 65-dB tone for 30 s, co-terminated with an unconditioned stimulus (US) of 0.5 mA, a 2-s foot shock.

Techniques: Over Expression, Injection, Virus, Expressing

Journal: Translational Psychiatry

Article Title: TOB is an effector of the hippocampus-mediated acute stress response

doi: 10.1038/s41398-022-02078-7

Figure Lengend Snippet: A Heatmaps for differentially expressed genes in hippocampi of Tob -KO compared to Tob -WT mice using RNA sequencing without fear conditioning training (naive) and 15 min, 1 h, 3 h after fear conditioning training (represented as z-scores of log raw counts, FC upregulated > 2 FC downregulated < 0.5, p < 0.05, FDR < 0.05). B Pathway analysis for RNA sequencing candidates using IPA software showing activation of hormonal concentration in hippocampus of Tob KO mice at 15 min post-conditioning. C Real-time PCR for lipocalin-2 ( Lcn2 ) mRNA in hippocampus of Tob -WT and KO naive mice and 15 min, 1 h and 3 h after fear conditioning. Two-way ANOVA followed by Bonferoni’s post-hoc test for multiple comparisons. D Western blotting showing protein expression of LCN-2 in hippocampi of naive Tob -KO mice and at 15 min, 1 h, and 3 h after fear conditioning training. E Normalized band intensity for LCN2 protein immunoblots. Two-way ANOVA followed by Bonferoni’s post-hoc test for multiple comparisons. F Western blotting showing abnormal protein expression in hippocampi of mice lacking Tob before and after fear conditioning training at 15 min, 1 h and 3 h. G Western blot band intensity quantification plots at different time points post-training compared to naive Tob -WT (p-ERK n = 4, MKP-1 n = 3). All values represent means ± SEMs. ns non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Briefly, on training day, mice were placed in a conditioning chamber (CL-3002L, O’Hara & Co Ltd., Japan) for 2 min to habituate, and then presented with a conditioning stimulus (CS) of a 65-dB tone for 30 s, co-terminated with an unconditioned stimulus (US) of 0.5 mA, a 2-s foot shock.

Techniques: RNA Sequencing, Software, Activation Assay, Concentration Assay, Real-time Polymerase Chain Reaction, Western Blot, Expressing