Journal: Journal of neuroinflammation

Article Title: Asrij/OCIAD1 depletion reduces inflammatory microglial activation and ameliorates Aβ pathology in an Alzheimer's disease mouse model.

doi: 10.1186/s12974-025-03415-5

Figure Lengend Snippet: Fig. 2 Asrij depletion reduces Aβ plaque deposition and associated pathology in APP/PS1 mice. A Representative confocal images of brain sections showing Thioflavin S (ThioS) staining (green) for plaques. DAPI (blue) marks the nuclei. Graphs show quantification of percent area covered by ThioS+ staining in hippocampus and cortex (n = 6, all males). B Representative confocal images of brain sections showing Aβ staining (red) using D54D2 antibody. DAPI (blue) marks the nuclei. Graphs show quantification of percent area covered by Aβ staining in hippocampus and cortex (n = 5, all males). C Graphs show quantification of Aβ42 levels (pg/mL) in the soluble (PBS) and insoluble (guanidine) brain fractions by ELISA (n = 6, all males). D Representative confocal images showing S100B (magenta) and GFAP (red) staining for astrocytes in the cortex. ThioS (green) and DAPI (blue) mark plaques and nuclei, respectively. Graph shows quantification of GFAP+ astrocytes per field of view (FOV) of the cortex averaged over 4 FOVs per section with 3 sections per mouse (n = 3 for WT and KO, n = 6 for AD and AD/KO, all males). E Representative confocal images showing LAMP1 staining (magenta) around ThioS+ Aβ plaques (green). Graph shows quantification of LAMP1+ area per plaque averaged over 15 plaques per section with 3 sections per mouse (n = 5, all males). F Representative confocal images showing APP staining (green) around 6E10+ Aβ plaques (red). Graph shows quantification of APP+ area per plaque averaged over 15 plaques per section with 3 sections per mouse (n = 3, all males). G Immunoblot analysis of PSD95 and Synaptophysin levels in the hippocampus. α-tubulin is loading control. Graphs show fold change in PSD95 and Synaptophysin levels normalized to α-tubulin (n = 6, all males). Statistical significance between experimental groups was calculated by unpaired two-tailed Student’s t-test (A-C and E-F) and one-way ANOVA with Tukey’s post hoc test (D and G). Error bars denote mean ± SEM. ns: non-significant, *P < 0.05, **P < 0.01, and ***P < 0.001

Article Snippet: Following primary antibodies were diluted in PBST and used at 4 °C overnight: APP (Abcam, #ab32136, 1:1000), OCIAD1 (Abcam, #ab91574, 1:1000), GFAP (Cloud-Clone Corp., #PAA068Mu02, 1:1000), IBA1 (Cell Signaling Technology, #17198S, 1:1000), CD68 (Cell Signaling Technology, #97778S, 1:1000), PSD95 (Cell Signaling Technology, #3450S, 1:1000), Synaptophysin (Cell Signaling Technology, #36406S, 1:2000), STAT3 (Cell Signaling Technology, #9139S, 1:1000), PhosphoSTAT3 (Tyr705) (Cell Signaling Technology, #9145S, 1:1000), NF-κB p65 (Cell Signaling Technology, #8242S, 1:1000), Phospho-NF-κB p65 (Ser536) (Cell Signaling Technology, #3033S, 1:1000), Akt (Cell Signaling Technology, #4691, 1:1000), Phospho-Akt (Ser243) (Cell Signaling Technology, #4060, 1:1000), mTOR (Abcam, #ab2732, 1:1000), Phospho-mTOR (Ser2448) (Abcam, #ab109268, 1:1000), GAPDH (Sigma-Aldrich, #G9545, 1:4000) and α-tubulin (Sigma-Aldrich, #T8203, 1:2000).

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Control, Two Tailed Test

Journal: Journal of neuropathology and experimental neurology

Article Title: Synaptophysin Is a Reliable Marker for Axonal Damage.

doi: 10.1093/jnen/nlw114

Figure Lengend Snippet: FIGURE 1. (A–C) Representative sections illustrate immunohistochemical staining for synaptophysin (Syp) in the cerebral cortex and hippocampus (A), cerebellum (B), and corpus callosum (C) of untreated control mice. (D) Accumulation of synaptophysin- positive vesicles ¼ spheroids/ovoids/bulbs in the corpus callosum of a cuprizone-treated mouse. Arrows indicate the different sizes of synaptophysin-positive-spheroids; small [S],<2 mm (as in panel C); middle-sized [M], 2–7.5 mm; large [L], 8–15 mm, pathological size. (E, F) Graphs show numbers of different sized synaptophysin-positive spheroids in the corpus callosum during acute (5 weeks, E) and chronic (12 weeks, F) demyelination followed by remyelination after stopping cuprizone. Significant effects between different time points were calculated separately for each type/size of bulbs (*p <0.05, **p < 0.01, **p < 0.001). Significant effects for different cuprizone time treatments in comparison to the untreated control: #p< 0.05, ##p < 0.01, ###p < 0.001), n ¼ 4–6. (G, H) Representative images show synaptophysin-positive bulbs in the demyelinated hippocampus (G) and internal capsule (H). (I-K) Representative images show that synaptophysin does not colocalize with the oligodendroglial marker Nogo-A (I), the astrocytic marker GFAP (J), or with the microglial marker Iba-1 (K). Scale bar in (K)¼ 100 mm, applies to (I) and (J).

Article Snippet: We used 2 different antibodies for synaptophysin: a mouse monoclonal IgG from Serotec and rabbit monoclonal from Abcam.

Techniques: Immunohistochemical staining, Staining, Control, Comparison, Marker

Journal: Journal of neuropathology and experimental neurology

Article Title: Synaptophysin Is a Reliable Marker for Axonal Damage.

doi: 10.1093/jnen/nlw114

Figure Lengend Snippet: FIGURE 2. (A, B) Graphs represent the quantitative analysis of APP, synaptophysin (Syp), and double-positive spheroids in the corpus callosum of cuprizone-treated mice with acute demyelination (A) and chronic demyelination (B). Two-way ANOVA analysis identify that there were no significant differences between numbers of APP- and synaptophysin-positive spheroids during

Article Snippet: We used 2 different antibodies for synaptophysin: a mouse monoclonal IgG from Serotec and rabbit monoclonal from Abcam.

Techniques:

Journal: Journal of neuropathology and experimental neurology

Article Title: Synaptophysin Is a Reliable Marker for Axonal Damage.

doi: 10.1093/jnen/nlw114

Figure Lengend Snippet: FIGURE 3. Immunohistochemical staining of the corpus callosum evaluating phosphorylation of neurofilaments and their colocalization with synaptophysin (Syp) during cuprizone-induced demyelination. (A–H) SMI-32 detecting non-phosphorylated heavy neurofilaments are shown in panels (A, C, E, G). SMI-312 detecting phosphorylated neurofilaments H and M are

Article Snippet: We used 2 different antibodies for synaptophysin: a mouse monoclonal IgG from Serotec and rabbit monoclonal from Abcam.

Techniques: Immunohistochemical staining, Staining, Phospho-proteomics

Journal: Journal of neuropathology and experimental neurology

Article Title: Synaptophysin Is a Reliable Marker for Axonal Damage.

doi: 10.1093/jnen/nlw114

Figure Lengend Snippet: FIGURE 5. (A-P) Representative sections show immunostaining in the corpus callosum for synaptophysin/Iba-1 (A-F) and synaptophysin/ PLP (G–L) during acute and chronic cuprizone induced demyelination and subsequent remyelination. Inserts in (A–F) show the morphology/shape of microglia identifying their activation state. Inserts in (G–L) show higher magnification of synaptophysin accumulations. Panels (M) and (N) are confocal images from RCA-1 (microglia activation marker, green) and synaptophysin (red) during inflammation in cuprizone-treated mice. Panels (O) and (P) show remyelinated axons (PLP, green) with internal synaptophysin accumulation (red).

Article Snippet: We used 2 different antibodies for synaptophysin: a mouse monoclonal IgG from Serotec and rabbit monoclonal from Abcam.

Techniques: Immunostaining, Activation Assay, Marker

Journal: Journal of neuropathology and experimental neurology

Article Title: Synaptophysin Is a Reliable Marker for Axonal Damage.

doi: 10.1093/jnen/nlw114

Figure Lengend Snippet: FIGURE 6. The appearance of synaptophysin (Syp)-positive pathological (medium-and large-sized) spheroids was compared and correlated with the extent of demyelination (proteolipid protein: PLP, myelin basic protein: MBP) and microglia infiltration (Iba-1) induced by cuprizone feeding (acute or chronic). (A, B) Graphs show the extent of microglia accumulation, appearance of pathological synaptophysin bulbs and the extent of de- and remyelination as judged by scoring of MBP and PLP immunostaining. Score of 3 represents complete myelination; score of 0 represents complete demyelination. (C–H) Correlation analyses of synaptophysin-positive bulbs with the extent of de- and remyelination and microglia numbers (n ¼ 5–6).

Article Snippet: We used 2 different antibodies for synaptophysin: a mouse monoclonal IgG from Serotec and rabbit monoclonal from Abcam.

Techniques: Immunostaining

Journal: Journal of neuropathology and experimental neurology

Article Title: Synaptophysin Is a Reliable Marker for Axonal Damage.

doi: 10.1093/jnen/nlw114

Figure Lengend Snippet: FIGURE 8. (A–C) Graphs show microglia/macrophage infiltration (A) and the course of demyelination (B) during Theiler’s virus induced encephalomyelitis in the thoracic segment of murine spinal cord. (C) Panel shows the appearance of pathologic accumulation of synaptophysin (Syp). Significant effects between different time points were indicated as: *p < 0.05, **p < 0.01, **p < 0.001. Significant effects for different time treatments in comparison to the untreated control are indicated as: # p < 0.05, ## p < 0.01, ### p < 0.001), n ¼ 4–6. (D–H) Representative images of thoracic spinal cord segment illustrate infiltration of microglia/macrophage at different days after the injection of Theiler’s murine encephalomyelitis virus. (I–U) Panel (I) shows the appearance of synaptophysin-positive bulbs exclusively in the demyelinated area; (J) and (K) illustrate the appearance of synaptophysin-positive spheroids surrounded by activated microglia/macrophages. (L–N) Colocalization of synaptophysin and APP in the spheroid structures. (O–U) Serial staining of the same spinal cord area during Theiler’s virus induced encephalomyelitis (O, synaptophysin/PLP double staining; P, synaptophysin/Iba-1 double staining; R, synaptophysin/APP double staining; S–U, higher magnification of area shown in R).

Article Snippet: We used 2 different antibodies for synaptophysin: a mouse monoclonal IgG from Serotec and rabbit monoclonal from Abcam.

Techniques: Virus, Comparison, Control, Injection, Staining, Double Staining

Journal: Journal of neuropathology and experimental neurology

Article Title: Synaptophysin Is a Reliable Marker for Axonal Damage.

doi: 10.1093/jnen/nlw114

Figure Lengend Snippet: FIGURE 9. Representative images show the expression of synaptophysin (Syp) in CNS tissue of 2 different patients with MS. (A– H) Different immunohistochemical stainings from a chronic active MS lesion (serial sections). Panels (E) and (F) show the higher magnifications from boxed areas 1 in (C) and (D). Panels (G) and (H) demonstrate higher magnification from boxed areas 2 in (C) and D. (I–K) Representative double immunostaining for synaptophysin and Iba-1 demonstrate microglia accumulation (I) and proving colocalization of APP and synaptophysin (J, K). (L–O) Demyelination/inflammation/axonal damage in another MS lesion. DAB staining in (L) and (M) illustrate the extent of demyelination (PLP) and inflammation (LN3 marks HLA-DR-positive cells: lymphocytes and macrophages) in this MS lesion. Panels N and O show higher magnification of the boxed are from this lesion in (L) and (M) demonstrating double expression of APP and synaptophysin in (N) and double staining of PLP and synaptophysin in (O). (P–U) Corpora amylacea are immunostain-positive for APP (P, S), and NeuN (U), but not synaptophysin (R, T).

Article Snippet: We used 2 different antibodies for synaptophysin: a mouse monoclonal IgG from Serotec and rabbit monoclonal from Abcam.

Techniques: Expressing, Immunohistochemical staining, Double Immunostaining, Staining, Double Staining