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