Journal: The Journal of experimental medicine

Article Title: Chronic TREM2 activation exacerbates Aβ-associated tau seeding and spreading.

doi: 10.1084/jem.20220654

Figure Lengend Snippet: Figure 1. Chronic TREM2 activation increases DAM around plaques. (A) Schematic of experimental design. 6-mo-old 5XFAD mice were injected with AD- tau in the HC (bregma: −2.5 mm; lateral: −2.0 mm; depth: −2.2 mm) and overlying cortex (bregma: −2.5 mm; lateral: −2.0 mm; depth: −1.0 mm) and sacrificed 3 mo later to evaluate peri-plaque pathologies. 1 wk before AD-tau injection and every week following until sacrifice, 5XFAD mice were given i.p. injections of 80 mg/ml of the AL002a mouse TREM2 antibody (n = 13 female, n = 14 male) or the IgG control antibody (n = 14 female, n = 12 male). (B) Quantification of TREM2 antibody levels in terminal plasma for 5XFAD mice either chronically treated with the IgG control antibody or the TREM2 antibody. (C) Quantification of TREM2 antibody levels in plasma for 5XFAD mice either acutely treated with the IgG control antibody or the TREM2 antibody. (D) Quantification of TREM2 antibody levels in cortical tissue for 5XFAD mice either acutely treated with the IgG control antibody or the TREM2 antibody. For acute treatment, there were n = 8 IgG control and n = 9 Trem2 antibody–treated age- and sex-matched mice, unless otherwise specified on graphs. (E) Representative images of ipsilateral hemisphere stained with CLEC7A+ microglia, P2RY12+ microglia, and Aβ from IgG control and TREM2 antibody–treated groups. Scale bars, 15 µm. (F) Quantification of mouse sTREM2 levels in plasma for 5XFAD mice either acutely treated with the IgG control antibody or the TREM2 antibody. (G) Quantification

Article Snippet: Biotinylated rat anti-TREM2 antibody (R&D Systems) was added at 1:10,000 dilution in binding buffer and incubated for 1 h at RT.

Techniques: Activation Assay, Injection, Control, Clinical Proteomics, Staining

Journal: The Journal of experimental medicine

Article Title: Chronic TREM2 activation exacerbates Aβ-associated tau seeding and spreading.

doi: 10.1084/jem.20220654

Figure Lengend Snippet: Figure 2. Chronic TREM2 activation with a TREM2 antibody results in no changes in Aβ plaque burden. (A) Representative images of Aβ plaques in 5XFAD mice either treated with the IgG control antibody or the TREM2 antibody. Scale bar, 100 µm. (B–E) Quantification of Aβ staining in the ipsi- and contralateral cortices (B and D) and hippocampi (C and E) in female mice. (F–I) Quantification of Aβ staining in the ipsi- and contralateral cortices (F and H) and hippocampi (G and I) in male mice. Triangle symbol represents female mice, and square symbol represents male mice either treated with the IgG control antibody (n = 14 female, n = 12 male) or the TREM2 antibody (n = 13 female, n = 14 male). Data are presented as mean ± SEM. Scale bar, 100 µm. Significance was determined using a Student’s t test. ns, P > 0.05.

Article Snippet: Biotinylated rat anti-TREM2 antibody (R&D Systems) was added at 1:10,000 dilution in binding buffer and incubated for 1 h at RT.

Techniques: Activation Assay, Control, Staining

Journal: The Journal of experimental medicine

Article Title: Chronic TREM2 activation exacerbates Aβ-associated tau seeding and spreading.

doi: 10.1084/jem.20220654

Figure Lengend Snippet: Figure 3. Chronic TREM2 activation with a TREM2 antibody increases NP-tau pathology. (A) Representative images of ipsi- and contralateral hemi- spheres stained with AT8+ NP-tau pathology in AD-tau injected 5XFAD mice either treated with the IgG control antibody or the TREM2 antibody. Repre- sentative images are from female mice. Scale bars, 100 µm. (B–E) Quantification of p-tau (AT8+) staining in the ipsi- and contralateral cortices (B and D) and hippocampi (C and E) in female mice. (F–I) Quantification of p-tau (AT8+) staining in the ipsi- and contralateral cortices (F and H) and hippocampi (G and I) in male mice. Triangle symbol represents female mice, and square symbol represents male mice either treated with the IgG control antibody (n = 14 female, n = 12 male) or the TREM2 antibody (n = 13 female, n = 14 male). Data are presented as mean ± SEM. Significance was determined using a Student’s t test. ns, P > 0.05; *, P < 0.05.

Article Snippet: Biotinylated rat anti-TREM2 antibody (R&D Systems) was added at 1:10,000 dilution in binding buffer and incubated for 1 h at RT.

Techniques: Activation Assay, Staining, Injection, Control

Journal: The Journal of experimental medicine

Article Title: Chronic TREM2 activation exacerbates Aβ-associated tau seeding and spreading.

doi: 10.1084/jem.20220654

Figure Lengend Snippet: Figure 4. Chronic TREM2 activation with a TREM2 antibody increases peri-plaque NP-tau pathology and plaque-associated neuritic dystrophy, and acute TREM2 activation results in no changes in AD-tau uptake and degradation. (A) Representative images of BACE1+ and X34+ staining in ipsilateral HC. Scale bars, 20 µm. (B) Representative images of AT8+ and X34+ staining in ipsilateral HC. (C–F) Quantification of the number of BACE1+ staining within 15 µm of plaques in the ipsi- and contra- cortices (C and E) and hippocampi (D and F). (G–J) Quantification of the number of AT8+ staining within 15 µm of plaques in the ipsi- and contra- cortices (G and I) and hippocampi (H and J). (K) AD-tau uptake assay in TREM2 WT BMDMs. Results represent two independent ex- periments. (L) AD-tau degradation assay in TREM2 WT BMDMs. Results represent two independent experiments. Triangle symbol represents female mice, and square symbol represents male mice either treated with the IgG control antibody (n = 14 female, n = 12 male) or the TREM2 antibody (n = 13 female, n = 14

Article Snippet: Biotinylated rat anti-TREM2 antibody (R&D Systems) was added at 1:10,000 dilution in binding buffer and incubated for 1 h at RT.

Techniques: Activation Assay, Staining, Degradation Assay, Control

Journal: The Journal of experimental medicine

Article Title: Chronic TREM2 activation exacerbates Aβ-associated tau seeding and spreading.

doi: 10.1084/jem.20220654

Figure Lengend Snippet: Figure 5. Chronic TREM2 activation with a TREM2 antibody increases peri-plaque loss of synaptic marker synapsin. (A) Representative images of Synapsin+ and X34+ staining in ipsilateral HC. Scale bar, 7 µm. (B) Quantification of the number of Synapsin+ puncta within 15 µm of plaques in the ipsi-HC. (C–E) Quantification of the number of Synapsin+ puncta within 15 µm of plaques in the ipsi-cortex, contra-cortex, and HC. Triangle symbol represents female mice, and square symbol represents male mice either treated with the IgG control antibody (n = 14 female, n = 12 male) or the TREM2 antibody (n = 13 female, n = 14 male). Data are presented as mean ± SEM. Significance was determined using a linear regression with sex as a covariate. ns, P > 0.05; *, P < 0.05.

Article Snippet: Biotinylated rat anti-TREM2 antibody (R&D Systems) was added at 1:10,000 dilution in binding buffer and incubated for 1 h at RT.

Techniques: Activation Assay, Marker, Staining, Control

Journal: Alzheimer's & Dementia : Translational Research & Clinical Interventions

Article Title: Novelty‐like activation of locus coeruleus protects against deleterious human pretangle tau effects while stress‐inducing activation worsens its effects

doi: 10.1002/trc2.12231

Figure Lengend Snippet: Phasic LC stimulation activates CD68 + microglia/macrophages. A1‐A5, IBa‐1 cell counts in the PC. Non‐E14: n = 5; E14: n = 7; E14‐P: n = 5; E14‐T: n = 6. B1‐B5, IBa‐1 cell counts in the DG. Non‐E14: n = 5; E14: n = 7; E14‐P: n = 5; E14‐T: n = 6. C1‐C5, CD‐68 cell counts in the PC. D1‐D5, CD‐68 cell counts in the DG. C‐D: Non‐E14: n = 5; E14: n = 8; E14‐P: n = 5; E14‐T: n = 5. Scale bars: 50 μm. * P < .05; ** P < .01. E‐H, Western blotting measuring levels of TREM2 (Non‐E14: n = 6; E14: n = 5; E14‐P: n = 6; E14‐T: n = 5; E), IL1a (Non‐E14: n = 7; E14: n = 7; E14‐P: n = 8; E14‐T: n = 6; F), MCHII (Non‐E14: n = 8; E14: n = 8; E14‐P: n = 8; E14‐T: n = 6; G), and BDNF (Non‐E14: n = 8; E14: n = 8; E14‐P: n = 8; E14‐T: n = 6; H) in the hippocampus. Data were normalized to Ponceau total protein staining

Article Snippet: The membranes were blocked for 2 hours with 5% non‐fat dry milk or BSA at room temperature, then incubated at 4°C overnight with the following antibodies: brain‐derived neurotrophic factor (BDNF; ab108319; Abcam, 1:2000), TREM2 (AF1729, R&D Systems, 1:5000), interleukin 1α (IL‐1α; OTI2F8, Novus Biologicals, 1:2000), human leukocyte antigen‐DR isotype (HLA‐DR; sc‐53319, Santa Cruz Biotechnology, 1:5000), anti‐phospho‐Tau (AT8; MN1020, Invitrogen, 1:1000), and paired helical filaments (PHF; MN1060, Invitrogen, 1:1000).

Techniques: Western Blot, Staining

Journal: Journal of Lipid Research

Article Title: The cholesteryl ester transfer protein (CETP) raises cholesterol levels in the brain

doi: 10.1016/j.jlr.2022.100260

Figure Lengend Snippet: CETP promotes ABCA7 and TREM2 expression in the liver. A: Feeding schedule and study design. Wt and CETP tg mice were fed for 3 months starting at the age of 2 months. Biochemical analyses were performed at 5 months. B–D: Normalized RT-qPCR from mouse liver tissue. B: H mgcr , C: L dlr , and D: L rp 1 expression, n = 6–14, mean ± SEM; 2-way ANOVA, Tukey’s multiple comparison test. E: Western blot analysis of ABCA7 and TREM2 from liver lysates. Antibodies used: rabbit-anti-GAPDH (14C10; Cell Signaling), anti-TREM2 (Mab1729; R&D Systems), and anti-ABCA7 (polyclonal; Thermo Fisher Scientific), n = 6; mean ± SEM, Student's t -test. F–I: Expression analysis of ABCA7 and TREM2 from liver samples. Normalized RT-qPCR levels of liver F: A bca 7 and H: T rem 2 ; n = 6–14, mean ± SEM; 2-way ANOVA, Tukey’s multiple comparison test. Western blot quantification of G: ABCA7 and I: TREM2; n = 6; mean ± SEM, Student's t -test.

Article Snippet: Primary antibodies used were 22C11 (Millipore), anti-GAPDH (14C10; Cell Signaling), TP2 (kind gift of the Ottawa Heart Institute), anti-triggering receptor expressed in myeloid cells 2 (TREM2) (Mab1729; R&D Systems) and anti-ABCA7 (polyclonal; Thermo Fisher Scientific), and horseradish peroxidase-coupled secondary antibodies (Promega).

Techniques: Expressing, Quantitative RT-PCR, Comparison, Western Blot

Journal: Journal of Neuroinflammation

Article Title: Genetic models of cleavage-reduced and soluble TREM2 reveal distinct effects on myelination and microglia function in the cuprizone model

doi: 10.1186/s12974-022-02671-z

Figure Lengend Snippet: Characterization of BMDM from wild-type, TREM2-IPD, TREM2-sol and TREM2-KO mice. a Schematic representation of the TREM2 receptor. Black, red and yellow asterisks indicate the cleavage site in WT, the mutated cleavage site in TREM2-IPD and in TREM2-sol, respectively. b Flow cytometry analysis of murine cell surface TREM2 on BMDM. MFI: median fluorescence intensity. Sheddase inhibitor: DCP333 (DPC), sheddase activator PMA. c Analysis of supernatants from b of murine soluble TREM2 from BMDM. d ATP-based cell survival assay of BMDM upon M-CSF deprivation for 2 and 3.5 days. ATP levels of cells cultured with M-CSF ( n = 7) were set as 100% survival and compared to the ATP concentration after 2 ( n = 4) and 3.5 days ( n = 3) without M-CSF for each genotype. Statistics: Holm–Sidak’s two-way ANOVA multiple comparisons (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001). e In vitro phagocytosis capacity of BMDM over 12 h (area-under-the curve) with 5 µg pHrodo-myelin per well ( n = 3). Fluorescence measurements in wells without prey were used as controls (data not shown). f Representative images of the Cathepsin B activity assay taken by the In-Cell Analyzer. The nuclei are stained with DAPI (blue), and the red fluorescence signals are derived from cleaved Magic red. g Quantification of the Cathepsin B assay images. The fluorescence integrated density of the Magic red signal was measured and normalized to the nuclei count. A significant ( p < 0.05) increase in normalized fluorescence between the DMSO control and K-18 within one genotype is marked by #. Statistics for d and g : Holm–Sidak’s two-way ANOVA with multiple comparisons (* p ≤ 0.05, ** p < 0.01, *** p < 0.001, *** p < 0.0001). Statistics for e : One-way ANOVA test with Holm–Sidak’s multiple comparisons test (*** p < 0.001; **** p < 0.0001). All data are presented as means ± SEM

Article Snippet: Next, the cells were transferred to a 96-well clear V-Bottom microplate and stained with a human/mouse TREM2 Alexa Fluor 488 antibody (R&D Systems FAB17291G) or a rat IgG2A Alexa Fluor ® 488-conjugated isotype control (R&D Sytems, IC006G), for 30 min at 4 °C.

Techniques: Flow Cytometry, Fluorescence, Clonogenic Cell Survival Assay, Cell Culture, Concentration Assay, In Vitro, Activity Assay, Staining, Derivative Assay, Control

Journal: Journal of Neuroinflammation

Article Title: Genetic models of cleavage-reduced and soluble TREM2 reveal distinct effects on myelination and microglia function in the cuprizone model

doi: 10.1186/s12974-022-02671-z

Figure Lengend Snippet: MRI indicated myelination deficits in TREM2-sol and TREM2-KO in the acute cuprizone model. a Schematic diagram of the experimental setup for the cuprizone treatment and recovery. Groups consisted of mice treated for 5 weeks with control food or 0.2% cuprizone in food and then switched back to control food (normal food) for the 4-week recovery. MRI measurements were performed at week 0 (baseline), week 3 (except for TREM2-KO and wt2) and week 5 of cuprizone intoxication, at week 7 (2 weeks of recovery on control food, except for TREM2-KO and wt2) and at week 9 (4 weeks of recovery on control food). Mice were culled at week 9 immediately after the last MRI measurement. b Representative MRI images acquired from three mice at baseline, at maximal pathology (5 weeks of receiving 0.2% cuprizone) and at recovery (4 weeks after switching to control food) for the different genotypes. c Corresponding T2-weighted MRI signal intensity (relative to the signal intensity at baseline) in external capsule (EC). Group sizes: n = 7–9 for all genotypes and timepoints. Data are shown as means ± SEM. Statistics: ANOVA with random effects comparisons indicated significant differences with respect to WT mice: *0.01 < p < 0.05, ***0.0001 < p < 0.001, **** p < 0.0001. For each group examined, T2-weighted signals were significantly increased with respect to baseline values (significances not shown). d Analysis of TREM2 levels in the brain for mice receiving control food (ctrl), at peak of cuprizone intoxication (week 5, cpz) and after 4-week recovery (rec). n.d. not detected. Statistics: Holm–Sidak’s multiple comparison test one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, ++++ p < 0.0001 to the respective wt group). Wt1, as well as wt ctrl1, wt cpz1 and wt rec1 are the respective wild-type groups for the study with TREM2-IPD and TREM2-sol, wt2 is the wild-type group for the TREM2-KO study

Article Snippet: Next, the cells were transferred to a 96-well clear V-Bottom microplate and stained with a human/mouse TREM2 Alexa Fluor 488 antibody (R&D Systems FAB17291G) or a rat IgG2A Alexa Fluor ® 488-conjugated isotype control (R&D Sytems, IC006G), for 30 min at 4 °C.

Techniques: Control, Comparison

Journal: Journal of Neuroinflammation

Article Title: Genetic models of cleavage-reduced and soluble TREM2 reveal distinct effects on myelination and microglia function in the cuprizone model

doi: 10.1186/s12974-022-02671-z

Figure Lengend Snippet: TREM2-sol and TREM2-KO display myelin debris, lack of remyelination and axonal pathology in the EC. Representative pictures for the different genotypes and timepoints from histological stainings detecting a myelin with Luxol Fast Blue (LFB) and corresponding quantitative optical density (OD) analysis of LFB in the EC (normalized to WT at control food), b mature oligodendrocytes (GST-π) and corresponding image analysis in EC (GST-π soma area in %), c myelin basic protein debris (dMBP) and corresponding image analysis in EC (dMBP-stained area in %), d neurofilament (SMI312) and corresponding image analysis in EC (SMI312-stained area in %). Group sizes: n = 3-7 for all genotypes and timepoints. Data shown as means ± SEM. wt: wild-type, TREM2-IPD: TREM2 cleavage-reduced, TREM2-sol: TREM2 soluble-only, TREM2-KO: TREM2 knockout. Ctrl: control food, cpz: cuprizone food for 5 weeks, rec: recovery on control food for 4 weeks. EC: external capsule, CC: corpus callosum. Scale bars: 300 µm (overview), 50 µm (close-up). Statistics: Holm–Sidak`s multiple comparison test one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). Comparisons not indicated are non-significant. Wt ctrl1, wt cpz1 and wt rec1 are the respective wild-type groups for the study with TREM2-IPD and TREM2-sol, Wt ctrl2, wt cpz2 and wt rec2 are the wild-type groups for the TREM2-KO study. Only statistical analysis within a study was performed. For c the analysis of the respective wt group for TREM2-KO was omitted as no dMBP signal was observed

Article Snippet: Next, the cells were transferred to a 96-well clear V-Bottom microplate and stained with a human/mouse TREM2 Alexa Fluor 488 antibody (R&D Systems FAB17291G) or a rat IgG2A Alexa Fluor ® 488-conjugated isotype control (R&D Sytems, IC006G), for 30 min at 4 °C.

Techniques: Control, Staining, Knock-Out, Comparison

Journal: Journal of Neuroinflammation

Article Title: Genetic models of cleavage-reduced and soluble TREM2 reveal distinct effects on myelination and microglia function in the cuprizone model

doi: 10.1186/s12974-022-02671-z

Figure Lengend Snippet: Increase of NF-L in plasma from TREM2-sol and TREM2-KO in the acute cuprizone model. NF-L measurements in plasma of WT, TREM2-IPD, TREM2-sol and TREM2-KO mice receiving control food (ctrl), at 5 weeks of cuprizone intoxication (cpz) and at 4-week recovery on normal food (rec). Group sizes: wt ctrl ( n = 4), wt cpz ( n = 4), wt rec ( n = 4), TREM2-IPD ctrl ( n = 3), TREM2-IPD cpz ( n = 7), TREM2-IPD rec ( n = 7), TREM2-sol ctrl ( n = 4), TREM2-sol cpz ( n = 7), TREM2-sol rec ( n = 4), TREM2-KO cpz ( n = 4). One-way ANOVA Holm–Šídák's multiple comparisons test, * p < 0.05, *** p < 0.001, **** p < 0.0001. Comparisons not indicated are non-significant. Wt ctrl1, wt cpz1 and wt rec1 are the respective wild-type groups for the study with TREM2-IPD and TREM2-sol, wt ctrl2, wt cpz2 and wt rec2 are the wild-type groups for the TREM2-KO study. Only statistical analysis within a study was performed

Article Snippet: Next, the cells were transferred to a 96-well clear V-Bottom microplate and stained with a human/mouse TREM2 Alexa Fluor 488 antibody (R&D Systems FAB17291G) or a rat IgG2A Alexa Fluor ® 488-conjugated isotype control (R&D Sytems, IC006G), for 30 min at 4 °C.

Techniques: Clinical Proteomics, Control

Journal: Journal of Neuroinflammation

Article Title: Genetic models of cleavage-reduced and soluble TREM2 reveal distinct effects on myelination and microglia function in the cuprizone model

doi: 10.1186/s12974-022-02671-z

Figure Lengend Snippet: TREM2-IPD and TREM2-sol mice show both sustained microglia/astrocyte activation and enhanced LAMP-1 in the EC. Representative images for the different genotypes and timepoints from histological stainings detecting a Iba1 and corresponding image analysis of Iba1-positive soma numbers (normalized to WT at week 5 cuprizone), b astrocytes (GFAP) and corresponding image analysis (GFAP-stained area in %), c LAMP-1 (lysosomal-associated membrane protein 1) and corresponding image analysis (LAMP1-stained area in %), as well as d TMEM119 (homeostatic marker) and corresponding image analysis (TMEM119-stained area in %). Group sizes: n = 2-7 for all genotypes and timepoints. Data are shown as means ± SEM. WT: wild-type, TREM2-IPD: TREM2 cleavage-reduced, TREM2-sol: TREM2 soluble-only, TREM2-KO: TREM2 knockout. Ctrl: control food, cpz: cuprizone food for 5 weeks, rec: recovery on control food for 4 weeks. EC: external capsule. CC: corpus callosum. Scale bars: 300 µm (overview), 50 µm (close-up). Statistics: Holm–Sidak’s multiple comparison test one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). Comparisons not indicated are non-significant. wt ctrl1, wt cpz1 and wt rec1 are the respective wild-type groups for the study with TREM2-IPD and TREM2-sol, wt ctrl2, wt cpz2 and wt rec2 are the wild-type groups for the TREM2-KO study. Only statistical analysis within a study was performed. For d the analysis of the respective wt group for TREM2-KO was omitted as no relevant TMEM119 signal was observed

Article Snippet: Next, the cells were transferred to a 96-well clear V-Bottom microplate and stained with a human/mouse TREM2 Alexa Fluor 488 antibody (R&D Systems FAB17291G) or a rat IgG2A Alexa Fluor ® 488-conjugated isotype control (R&D Sytems, IC006G), for 30 min at 4 °C.

Techniques: Activation Assay, Staining, Membrane, Marker, Knock-Out, Control, Comparison

Journal: Journal of Neuroinflammation

Article Title: Genetic models of cleavage-reduced and soluble TREM2 reveal distinct effects on myelination and microglia function in the cuprizone model

doi: 10.1186/s12974-022-02671-z

Figure Lengend Snippet: Brain cytokine/chemokine response is reduced in TREM2-sol and TREM2-KO, but enhanced in TREM2-IPD. Cytokine/chemokine measurements in brain detecting MIP-1a, MIP-1b, IP-10 and MCP-1 in WT, TREM2-IPD, TREM2-sol and TREM2-KO with control food (ctrl), at 5 weeks of cuprizone intoxication (cpz) and at 4-week recovery on normal food (rec). Measurements are normalized to wt cpz. Group sizes: wt ctrl ( n = 4), wt cpz ( n = 4), wt rec ( n = 4), TREM2-IPD ctrl ( n = 4), TREM2-IPD cpz ( n = 7), TREM2-IPD rec ( n = 7), TREM2-sol ctrl ( n = 4), TREM2-sol cpz ( n = 7), TREM2-sol rec ( n = 4), TREM2-KO ctrl ( n = 7), TREM2-KO cpz ( n = 4), TREM2-KO rec ( n = 7). Statistics: ordinary one-way ANOVA Holm–Šídák's multiple comparisons test, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Comparisons not indicated are non-significant. wt ctrl1, wt cpz1 and wt rec1 are the respective wild-type groups for the study with TREM2-IPD and TREM2-sol, wt ctrl2, wt cpz2 and wt rec2 are the wild-type groups for the TREM2-KO study. Only statistical analysis within a study was performed

Article Snippet: Next, the cells were transferred to a 96-well clear V-Bottom microplate and stained with a human/mouse TREM2 Alexa Fluor 488 antibody (R&D Systems FAB17291G) or a rat IgG2A Alexa Fluor ® 488-conjugated isotype control (R&D Sytems, IC006G), for 30 min at 4 °C.

Techniques: Control

Journal: Journal of Neuroinflammation

Article Title: Genetic models of cleavage-reduced and soluble TREM2 reveal distinct effects on myelination and microglia function in the cuprizone model

doi: 10.1186/s12974-022-02671-z

Figure Lengend Snippet: TREM2-IPD showed enhanced myelination in the chronic cuprizone model. a Schematic diagram of the experimental setup for the cuprizone treatment and recovery. Groups consisted of mice treated for 12 weeks with control food (normal food) or 0.2% cuprizone in food and then switched back to control food for the 3-week recovery. MRI measurements were performed at timepoints indicated. Mice were culled at week 15 immediately after the last MRI measurement. b T2-weighted signals in the CC and EC during the 12-week intoxication period and the recovery phase were significantly increased with respect to baseline values and compared to analyses for mice receiving control diet throughout the experiment. The significance levels # 0.01 < p < 0.05 and ### p < 0.001 correspond to ANOVA with random effects comparisons between WT and TREM2-IPD animals. Representative images for the different genotypes and at week 15 from histological stainings detecting c myelin with Luxol Fast Blue (LFB) and corresponding quantitative optical density analysis of LFB in the EC and CC, and d mature oligodendrocytes (GST-π) and corresponding image analysis in EC and CC (GST-π soma area in %). Group sizes: n = 5–7 for all genotypes and timepoints. Male mice were used for the cuprizone groups. Data are shown as means ± SEM. WT: wild-type, TREM2-IPD: TREM2 cleavage-reduced. Ctrl: control food, rec: recovery on control food for 3 weeks. Control refers to TREM2-IPD mice receiving normal food throughout the study. EC: external capsule, CC: corpus callosum. Scale bars: 500 µm. Statistics: ordinary one-way ANOVA Holm–Šídák’s multiple comparisons test, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: not significant

Article Snippet: Next, the cells were transferred to a 96-well clear V-Bottom microplate and stained with a human/mouse TREM2 Alexa Fluor 488 antibody (R&D Systems FAB17291G) or a rat IgG2A Alexa Fluor ® 488-conjugated isotype control (R&D Sytems, IC006G), for 30 min at 4 °C.

Techniques: Control

Journal: Brain, behavior, and immunity

Article Title: Early life stress impairs synaptic pruning in the developing hippocampus

doi: 10.1016/j.bbi.2022.09.014

Figure Lengend Snippet: ELS reduces expression of TREM2 and CD11b while increasing CX3CR1 in P17 microglia. (A) Scatter plots for quantifying Trem2 positive microglia in CTL, LB and UPS. (B) Percentage of TREM2-positive microglia. Median fluorescence intensity (MFI) plots and surface expression of CD11b (C-D), CD45 (E-F) and CX3CR1 (G-H). CTL-males: n = 16, CTL-females: n = 14, LB males: n = 10, LB-females: n = 9, UPS-males: n = 12, UPS-females: n = 16. Error bars represent mean ± SEM. **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Microglia were isolated from the hippocampus as described above and blocked with rat- anti CD16/CD32 Fc antibodies 1:100 (Cat# 553142, BD Biosciences) and then stained with rat-anti CD11b-PeCy7 1:100 (Cat# 25-0112-82, ebiosciences), rat-anti CD45-PeCPCy5.5 antibodies 1:100 (Cat# 45-0451-82, ebiosciences), APC-rat anti Trem2 antibodies 1:10 (Cat# FAB17291A R & D systems), or APC-mouse anti CX3CR1 antibodies 1:100 (Cat# 149008) for 25 min on ice.

Techniques: Expressing, Fluorescence

Journal: Brain, behavior, and immunity

Article Title: Early life stress impairs synaptic pruning in the developing hippocampus

doi: 10.1016/j.bbi.2022.09.014

Figure Lengend Snippet: Trem2 is essential for normal phagocytic activity. (A-B) Effects of Trem2 genotype (WT, Hets, Ko) on ex vivo phagocytic activity in microglia isolated from the hippocampus of 17-day old pups (n = 10–17 pups per group, 50 % females). (C) Representative confocal images (top row) and Imaris models (middle and lower rows) of microglia from Trem2 wildtype (WT), heterozygous (Hets) and knockout P17 littermates. Staining for Iba1 (green), CD68 (blue), and PSD95 (red). Middle row: reconstruction of Iba1 & CD68 staining. Lower row: reconstruction of CD68 and PSD95 staining inside microglia. Effects of Trem2 genotype on microglial volume (D), CD68 volume inside microglia (E), number of PSD95 puncta in microglia (F) and PSD95 puncta inside CD68 (G), n = 5 cells per mouse and 4–5 mice per group, 50 % females). Error bars represent mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Microglia were isolated from the hippocampus as described above and blocked with rat- anti CD16/CD32 Fc antibodies 1:100 (Cat# 553142, BD Biosciences) and then stained with rat-anti CD11b-PeCy7 1:100 (Cat# 25-0112-82, ebiosciences), rat-anti CD45-PeCPCy5.5 antibodies 1:100 (Cat# 45-0451-82, ebiosciences), APC-rat anti Trem2 antibodies 1:10 (Cat# FAB17291A R & D systems), or APC-mouse anti CX3CR1 antibodies 1:100 (Cat# 149008) for 25 min on ice.

Techniques: Activity Assay, Ex Vivo, Isolation, Knock-Out, Staining