Journal: Fluids and Barriers of the CNS

Article Title: Age‑dependent and post‑intraventricular hemorrhage remodeling of the ependymal glycocalyx in mice

doi: 10.1186/s12987-025-00725-x

Figure Lengend Snippet: Visualization and glycan profiling of the ependymal Gcx in young adult mice. ( a ) Scanning electron microscopy (SEM) images of ventricular ependymal cells. Gcx is not visible under conventional glutaraldehyde fixation (left) but clearly visualized surrounding the cell surface and cilia with lanthanum staining (right). ( b ) Alcian blue + hematoxylin-eosin (HE) staining. Glycans covering the apical surface of the ependyma are stained blue with Alcian blue. White scale bar: 20 μm. ( c ) Low-vacuum SEM image of a serial section corresponding to ( b ). A three-dimensional view of the Gcx layer covering the base of the cilia and surrounding the ciliary shafts at the apical surface. Enlarged view on the right. ( d ) Double immunofluorescence staining of lectin (PNA, red) and S100β (green) in frozen sections. A distinct glycan layer is observed at the apical surface of the ependyma, comparable to that seen in electron microscopy. White scale bar: 10 μm. ( e ) Fluorescence intensity plot along the measurement line (yellow) in ( d ), from the ventricular lumen (left) toward the brain parenchyma (right); red indicates lectin, green indicates S100β. The plot confirms localization of the Gcx at the ependymal cell surface. ( f ) Bar graph showing the mean fluorescence intensity of the ependymal Gcx for each of the 21 lectins examined ( n = 3 mice; total of 30 cells). ( g ) Merged images of lectin (red) and S100β (green) staining. Representative examples are shown: strong positive (RCA-I, STL), moderate positive (WGA, PHA-E), and negative (UEA-I, S-WGA). White scale bar: 10 μm

Article Snippet: Subsequently, biotinylated lectins (1:200 dilution) and an ependymal cell marker, the S100β antibody (mouse monoclonal, 1:500 dilution; sc-393919, Santa Cruz Biotechnology), were applied and incubated overnight at 4 °C.

Techniques: Glycoproteomics, Electron Microscopy, Staining, Double Immunofluorescence Staining, Fluorescence

Journal: Fluids and Barriers of the CNS

Article Title: Age‑dependent and post‑intraventricular hemorrhage remodeling of the ependymal glycocalyx in mice

doi: 10.1186/s12987-025-00725-x

Figure Lengend Snippet: Comparison of ependymal Gcx between young adult and aged mice. ( a ) Heatmap showing the mean fluorescence intensity per pixel of the ependymal glycocalyx (Gcx) for 21 lectins in young adult and aged mice ( n = 3 mice per group; 10 cells per mouse). ( b ) Representative PNA-based fluorescence images of the ependymal Gcx in young adult (top) and aged (bottom) mice. White scale bar, 5 μm. ( c ) Quantification of Gcx coverage along the periventricular circumference of the lateral ventricle ( n = 3 mice per group; two-sided t-test; mean ± S.E.M.). ( d ) Transmission electron microscopy (TEM) images of ependymal cells. In young adults (left), the Gcx is visible on the apical surface (black arrowheads), microvilli (black arrows), and cilia (small black arrow). In aged mice (right), loss of the Gcx is evident on the apical surface (white arrowheads) and microvilli (white arrows). White scale bar, 500 nm. ( e ) Schematic illustration of the ependymal surface Gcx based on lectin staining and TEM. ( f ) Double fluorescence staining for LEL (red) and S100β (green) (left) with corresponding fluorescence intensity profiles (right; red = LEL, green = S100β; dots indicate inflection points). White scale bar, 5 μm. ( g ) Correlation between inflection point distance and manually measured Gcx thickness using PNA staining ( n = 20 measurement sites; Pearson correlation). ( h ) Comparison of Gcx thickness between young adult and aged mice determined by inflection point distances for LEL, PNA, and RCA-I ( n = 3 mice per group; 16 sites per mouse; two-sided t-test; mean ± S.E.M.). ( i ) Representative double fluorescence staining for PNA (red) and S100β (green) showing enhanced cytoplasmic PNA positivity in aged ependymal cells. White scale bar, 5 μm. ( j ) Cytoplasmic PNA-positive rate along the ventricular wall ( n = 3 per group; two-sided t-test; mean ± S.E.M.). ( k ) Heatmap of mean fluorescence intensity in the ependymal cytoplasm for 21 lectins ( n = 3 per group; 10 cells per mouse)

Article Snippet: Subsequently, biotinylated lectins (1:200 dilution) and an ependymal cell marker, the S100β antibody (mouse monoclonal, 1:500 dilution; sc-393919, Santa Cruz Biotechnology), were applied and incubated overnight at 4 °C.

Techniques: Comparison, Fluorescence, Transmission Assay, Electron Microscopy, Staining

Journal: Gut Microbes

Article Title: Gut microbial ammonia as a mediator of PFOS neurotoxicity and its remediation by the flavonoid Icaritin

doi: 10.1080/19490976.2026.2620125

Figure Lengend Snippet: ICT alleviates PFOS-induced activation of hippocampal astrocytes and Aβ pathology, neuroinflammation, and oxidative stress. (A) Representative H&E staining of the hippocampal dentate gyrus (DG) region showing neuronal morphology. Scale bars: 500 μm (upper panel, overview), 50 μm (lower panel, detailed view). (B) Dual immunofluorescence staining of GFAP (green, astrocyte marker) and Aβ (red) in the hippocampal DG region. (C) Quantitative analysis of GFAP fluorescence intensity ( n = 5). (D) Quantitative analysis of Aβ fluorescence intensity ( n = 5). (E) Triple-label immunofluorescence staining of GFAP (green), C3 (red) and S100A10 (purple) in the hippocampal DG region. (F) Quantitative analysis of C3 + GFAP + cells (%) ( n = 5). (G) Quantitative analysis of S100A10 + GFAP + cells (%) ( n = 5). (H−J) Hippocampal levels of pro-inflammatory cytokines: (H) IL-1β ( n = 6), (I) IL-6 ( n = 6), and (J) TNF- α ( n = 6), measured by ELIZA. (K) Reactive oxygen species (ROS) levels in hippocampal tissues ( n = 6). (L) Malondialdehyde (MDA) content, a lipid peroxidation marker ( n = 6). (M−O) Activities of antioxidant enzymes: (M) catalase (CAT, n = 6), (N) superoxide dismutase (SOD, n = 6), and (O) glutathione peroxidase (GSH-Px, n = 6) in hippocampal tissues. The data were presented as the mean ± SEM. ** P < 0.01 vs. Control group; # P < 0.05, ## P < 0.01 vs. PFOS group.

Article Snippet: GFAP (16825-1-AP), C3 (21337-1-AP), S100A10 (11250-1-AP) and β -tubulin (10094-1-AP) were purchased from Proteintech (Wuhan, China). β -amyloid (Aβ 1-42 ) peptide was purchased from Macklin (Shanghai, China).

Techniques: Activation Assay, Staining, Immunofluorescence, Marker, Fluorescence, Control