Journal: Frontiers in Cellular Neuroscience

Article Title: Glial D-serine modulates oligodendrocyte lineage progression under inflammatory conditions

doi: 10.3389/fncel.2026.1784678

Figure Lengend Snippet: Temporal progression and cellular composition of primary OPC cultures. Phase-contrast images illustrate OPC morphology at DIV1, DIV5, and DIV7 (A) . Quantification of OLIG2 + and MBP + cells across days in vitro (DIV1–DIV7), expressed as percentage of total DAPI + nuclei per field, is shown in (B) , while total DAPI + nuclei per field were quantified over time as an estimate of net cell number (C) . Representative immunofluorescence images at DIV7 display OLIG2 + or MBP + cells (green) with DAPI counterstaining (blue) (D) . Culture purity was evaluated at DIV7 by immunostaining for PDGFRα (green) together with microglial (IBA1; panel E , red) or astrocytic markers (GFAP; panel F , red). Representative fields are shown to illustrate residual non-oligodendroglial cells; quantitative analysis across multiple fields confirmed minimal microglial and astrocytic contamination (<1% of total DAPI + nuclei). Data represent mean ± SEM from seven independent cultures derived from pooled neonatal cortices. Scale bars: 50 μm (A,D) ; 100 μm (E,F) .

Article Snippet: The following primary antibodies were used to identify distinct stages of the oligodendrocyte lineage: platelet-derived growth factor receptor alpha (PDGFRα; rabbit monoclonal, clone D13C6 XP®, Alexa Fluor® 488–conjugated, Cell Signaling Technology, Cat. No. 8871, 1:200) for OPCs; Olig2 (goat polyclonal, R&D Systems, Cat. No. AF2418, 5–15 μg/mL) for oligodendroglial lineage cells; and myelin basic protein (MBP; rabbit monoclonal, clone D8X4Q XP®, Cell Signaling Technology, Cat. No. 78896, 1:50) for mature oligodendrocytes.

Techniques: In Vitro, Immunofluorescence, Immunostaining, Derivative Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Glial D-serine modulates oligodendrocyte lineage progression under inflammatory conditions

doi: 10.3389/fncel.2026.1784678

Figure Lengend Snippet: D-serine attenuates late-stage oligodendrocyte lineage progression without altering net cell number. Representative immunofluorescence images of OPC cultures treated with D-serine (1 or 10 μM) at DIV6 and analyzed at DIV7 are shown for MBP (A) and OLIG2 (C) , with DAPI used to label nuclei. Merged images illustrate lineage marker distribution across conditions. The proportions of MBP + (B) and OLIG2 + cells (D) are expressed as a percentage of total DAPI + nuclei per field. (E) Total DAPI + nuclei per field are shown normalized to control values and used as an estimate of net cell number. (F) The proportion of OLIG2 − /MBP − cells and (G) the proportion of PDGFRα + cells are expressed as a percentage of total DAPI + nuclei per field. Bars represent mean ± SEM from 6 to 10 independent cultures derived from pooled neonatal cortices. Statistical comparisons were performed using the Kruskal–Wallis test followed by Dunn’s post hoc test. * p < 0.05; ** p < 0.01.

Article Snippet: The following primary antibodies were used to identify distinct stages of the oligodendrocyte lineage: platelet-derived growth factor receptor alpha (PDGFRα; rabbit monoclonal, clone D13C6 XP®, Alexa Fluor® 488–conjugated, Cell Signaling Technology, Cat. No. 8871, 1:200) for OPCs; Olig2 (goat polyclonal, R&D Systems, Cat. No. AF2418, 5–15 μg/mL) for oligodendroglial lineage cells; and myelin basic protein (MBP; rabbit monoclonal, clone D8X4Q XP®, Cell Signaling Technology, Cat. No. 78896, 1:50) for mature oligodendrocytes.

Techniques: Immunofluorescence, Marker, Control, Derivative Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Glial D-serine modulates oligodendrocyte lineage progression under inflammatory conditions

doi: 10.3389/fncel.2026.1784678

Figure Lengend Snippet: Conditioned media from LPS-stimulated glial cultures modulate OPC differentiation in a D-serine- and NMDAR-dependent manner. Representative immunofluorescence images illustrate MBP (A) , OLIG2 (C) , and combined MBP/TUNEL staining (E) in OPC cultures treated with conditioned media (CM) derived from mixed glial cultures exposed to LPS for 1 h, with or without enzymatic degradation of D-serine using DAAO/catalase (DAAO/CAT) or NMDAR antagonism with MK-801. DAPI was used to label nuclei. All images were acquired using identical imaging settings across conditions. Scale bar: 50 μm (applies to panels A , C , and E ). (B) Percentage of MBP + cells in OPC cultures. (D) Percentage of OLIG2 + cells. (F) Apoptotic MBP + cells quantified by TUNEL staining. For (F) , data are normalized to the mean value observed in the NT group (non-treated OPC cultures maintained without conditioned media; 14.9% ± 1.6% of total cells), and no significant differences were detected across treatments. Data are presented as mean ± SEM from 6 to 9 independent cultures. Statistical comparisons were performed using the Kruskal–Wallis test followed by Dunn’s post hoc test. * p < 0.05; # p < 0.05 compared to NT.

Article Snippet: The following primary antibodies were used to identify distinct stages of the oligodendrocyte lineage: platelet-derived growth factor receptor alpha (PDGFRα; rabbit monoclonal, clone D13C6 XP®, Alexa Fluor® 488–conjugated, Cell Signaling Technology, Cat. No. 8871, 1:200) for OPCs; Olig2 (goat polyclonal, R&D Systems, Cat. No. AF2418, 5–15 μg/mL) for oligodendroglial lineage cells; and myelin basic protein (MBP; rabbit monoclonal, clone D8X4Q XP®, Cell Signaling Technology, Cat. No. 78896, 1:50) for mature oligodendrocytes.

Techniques: Immunofluorescence, TUNEL Assay, Staining, Derivative Assay, Imaging

Journal: Frontiers in Neurology

Article Title: Subregional differences in the hippocampal transcriptomic response after penetrating traumatic brain injury in rats

doi: 10.3389/fneur.2025.1729794

Figure Lengend Snippet: Schematic overview of hippocampal subregion sampling and transcriptional responses following TBI. Schematic illustration showing the hippocampal section (Bregma −3.6 mm) and the standardized subregional sampling locations [CA1 (a) , CA2 (b) , CA3 (c) , DG (d) ] used for laser-capture microdissection (LCMD). Representative immunofluorescence images of the captured regions (a–d) show local cellular composition with DAPI (nuclei, blue), NeuN (neurons, green), GFAP (astrocytes, orange), and Olig2 (oligodendrocytes, red) labeling, confirming clear neuronal layer delineation and low glial contamination, primarily consisting of astrocytic processes. Adjacent panels display corresponding volcano plots illustrating differential gene expression (DGE) between TBI and sham for each subregion. Analyses were performed using DESeq2 v1.44.0 with log 2 fold-change shrinkage (apeglm) and Benjamini–Hochberg multiple-testing correction. Genes with adjusted p < 0.05 and log 2 fold-change > 1 were considered significant. The x -axis represents log₂ fold change, and the y -axis shows –log₁₀ adjusted p -values, with red and blue points indicating significantly upregulated and downregulated genes, respectively. Across subregions, a predominance of upregulated genes was observed, with the most extensive transcriptional activation in CA2 and CA3, reflecting pronounced subregion-specific injury responses. LCMD was consistently performed at the same anatomical coordinates in each animal to ensure regional comparability. The image was partially created in BioRender. CA, cornu ammonis; DG, dentate gyrus; DGE, differential gene expression; LCMD, laser-capture microdissection; TBI, traumatic brain injury.

Article Snippet: Slides were incubated with blocking solution (0.01 M PBS + 0.1% NaN3 + 0.3% Triton + 5% BSA) for 45 min at room temperature, followed by incubation with primary antibodies toward microglia (rabbit anti-Iba1, FUJIFILM, cat no: 019–19,741, 1:500), neurons (chicken anti-NeuN, Sigma, cat no: ABN91, 1:1000), astrocytes (rabbit anti-GFAP, Abcam, cat no: ab33922, 1:1000), oligodendrocytes (goat anti-OLIG2, R&D Systems, AF2418, 1:500), leukocytes (mouse anti-CD45, Abcam, cat no: ab33923, 1:50), oligodendrocyte precursors (mouse anti-NG2, Millipore, cat no: MAB5384-I, 1:50), epithelial tight junctions/vessels (rabbit anti-Occludin, Abcam, cat no: ab216327, 1:200), in blocking solution for 16 h at 4 °C.

Techniques: Sampling, Laser Capture Microdissection, Immunofluorescence, Labeling, Gene Expression, Activation Assay

Journal: Frontiers in Neurology

Article Title: Subregional differences in the hippocampal transcriptomic response after penetrating traumatic brain injury in rats

doi: 10.3389/fneur.2025.1729794

Figure Lengend Snippet: Cellular composition of hippocampal subregions surrounding the pyramidal and granule cell layers 72 h post-injury. Representative immunofluorescence images (20 × magnification, 2 × 2 tile scans) from hippocampal subfields (CA1–CA3, DG) located at Bregma −3.6 mm from animals subjected to penetrating TBI. Sections were stained for DAPI (nuclei), NeuN (neurons), GFAP (astrocytes), Olig2 (oligodendrocytes), NG2 (oligodendrocyte progenitors), Occludin (endothelial tight junctions/vessels), Iba1 (microglia), and CD45 (infiltrating immune cells). The aim was to assess local cellular composition around the pyramidal and granule cell layers and evaluate potential non-neuronal contamination in the laser-captured regions. NeuN staining confirmed the strong neuronal specificity of these layers, with only sparse GFAP-positive processes suggesting low astrocytic contribution. No Olig2-, NG2-, Iba1-, or CD45-positive cells were detected within the neuronal layers, indicating negligible contamination by oligodendrocytes, OPCs, microglia, or infiltrating immune cells. Occludin labeling was confined to perivascular regions outside the dissected zones. Images are representative of areas used for laser-capture microdissection and derived from immediately adjacent 14 μm sections. Larger images are available in . CA, cornu ammonis; DG, dentate gyrus; TBI, traumatic brain injury.

Article Snippet: Slides were incubated with blocking solution (0.01 M PBS + 0.1% NaN3 + 0.3% Triton + 5% BSA) for 45 min at room temperature, followed by incubation with primary antibodies toward microglia (rabbit anti-Iba1, FUJIFILM, cat no: 019–19,741, 1:500), neurons (chicken anti-NeuN, Sigma, cat no: ABN91, 1:1000), astrocytes (rabbit anti-GFAP, Abcam, cat no: ab33922, 1:1000), oligodendrocytes (goat anti-OLIG2, R&D Systems, AF2418, 1:500), leukocytes (mouse anti-CD45, Abcam, cat no: ab33923, 1:50), oligodendrocyte precursors (mouse anti-NG2, Millipore, cat no: MAB5384-I, 1:50), epithelial tight junctions/vessels (rabbit anti-Occludin, Abcam, cat no: ab216327, 1:200), in blocking solution for 16 h at 4 °C.

Techniques: Immunofluorescence, Staining, Labeling, Laser Capture Microdissection, Derivative Assay