Journal: bioRxiv

Article Title: Transcriptional Heterogeneity Reveals a Synaptic Gene Program in Developing and Adult Human Oligodendrocyte Precursor Cells

doi: 10.64898/2026.01.01.697017

Figure Lengend Snippet: (A) Human embryonic (PCW 17) OPCs were isolated based on PDGFRα immunoreactivity (see Methods) and subjected to scRNA-seq. Graph clustering based on transcriptomic similarity revealed four OPC clusters ( e mbryonic clusters 1-4) in UMAP space. (B) Cells showed robust expression of OLIG2 and PDGFRα , confirming OPC identity. (C) Mature oligodendrocyte markers MAG or MOG were low. (D) Heatmap representation of a unique and largely non-overlapping gene expression signatures for each OPC cluster. (E-H) Top seven pathways identified by Gene Ontology (GO) analysis of differentially expressed ( P < 0.01) genes in each OPC cluster.

Article Snippet: To isolate eSyn-OPCs, PDGFRα + OPCs were passed on to a dish coated with GRB14 antibody (Proteintech, 15298-1-AP) for 20min at 37C.

Techniques: Isolation, Expressing, Gene Expression

Journal: bioRxiv

Article Title: Transcriptional Heterogeneity Reveals a Synaptic Gene Program in Developing and Adult Human Oligodendrocyte Precursor Cells

doi: 10.64898/2026.01.01.697017

Figure Lengend Snippet: (A) (Top left) Graph clustering of eSyn-OPCs in UMAP space from the PCW 17 brain, highlighted in red. (B) eSyn-OPCs express known synaptic development genes, including THBS2, PLAT, WNT5A, WNT7A, and ACHE .

Article Snippet: To isolate eSyn-OPCs, PDGFRα + OPCs were passed on to a dish coated with GRB14 antibody (Proteintech, 15298-1-AP) for 20min at 37C.

Techniques:

Journal: bioRxiv

Article Title: Transcriptional Heterogeneity Reveals a Synaptic Gene Program in Developing and Adult Human Oligodendrocyte Precursor Cells

doi: 10.64898/2026.01.01.697017

Figure Lengend Snippet: (A) A representative cortical section with DAPI staining from a PCW 22 brain. (B) Immunohistochemistry for eSyn-OPCs using the eSyn-OPC marker HAPLN1 and colocalizing with PDGFRα. Scale bar unit = um. (C) Quantification of HAPLN1 + /PDGFRα + eSyn-OPCs revealed that the appearance of eSyn-OPCs occurs after PCW 12, and the majority of eSyn-OPCs are found in layers IZ, oSVZ, iSVZ, and VZ. (D) Visium spatial transcriptomics from a PCW 15 sample. (E) K-means clustering of 1,880 spots from (D) identified five distinct layers in UMAP space. (F) Quantification of MT1E + eSyn-OPC spots in the five cortical layers found in (E). (G) Quantification of PDGFRα + OPC spots in the five cortical layers found in (E). (H) The mRNA levels of neural progenitor ( EOMES ) and neural stem cell ( SOX2, ID4 ) markers in the MT1E + spots (containing eSyn-OPCs) compared to PDGFR α + / MT1E - spots (containing OPCs but not eSyn-OPCs). (I) Immunohistochemistry confirmed proximity of eSyn-OPCs and SOX2 + neural stem cells in PCW 22 cortical tissue. Scale bar unit = um.

Article Snippet: To isolate eSyn-OPCs, PDGFRα + OPCs were passed on to a dish coated with GRB14 antibody (Proteintech, 15298-1-AP) for 20min at 37C.

Techniques: Staining, Immunohistochemistry, Marker

Journal: bioRxiv

Article Title: Transcriptional Heterogeneity Reveals a Synaptic Gene Program in Developing and Adult Human Oligodendrocyte Precursor Cells

doi: 10.64898/2026.01.01.697017

Figure Lengend Snippet: (A) Identification of OPCs in the UMAP plot of brain cells obtained from a healthy 29-year-old male (from Siletti et al.). OPCs were defined by co-expression of marker genes PDGFRA and OLIG2 , and mature oligodendrocytes were defined by co-expression of marker genes MAG , MOG , and MBP . (B) Re-clustering of the OPC cluster found in (A) based on differentially expressed genes identified four OPC clusters ( a dult clusters 1-4). (C-F) Top seven pathways identified by GO analysis of differentially expressed genes in each OPC cluster from (B). aCluster 4 was identified as aSyn-OPCs.

Article Snippet: To isolate eSyn-OPCs, PDGFRα + OPCs were passed on to a dish coated with GRB14 antibody (Proteintech, 15298-1-AP) for 20min at 37C.

Techniques: Expressing, Marker

Journal: bioRxiv

Article Title: Transcriptional Heterogeneity Reveals a Synaptic Gene Program in Developing and Adult Human Oligodendrocyte Precursor Cells

doi: 10.64898/2026.01.01.697017

Figure Lengend Snippet: (A) A simulated null distribution of gene-set overlaps between eSyn-OPCs and randomly sampled gene sets of the same size expressed in the brain ( n = 10,000). Adult Syn-OPCs (aSyn-OPCs) showed a significant overlap (z = 3.04, p = 0.0011) with eSyn-OPCs, indicating genetic similarity between the two populations. (B) Among synapse-related genes, eSyn-OPCs showed a greater abundance of genes regulating synapse structure compared to aSyn-OPCs. (C) SynGO, a synaptic gene analysis tool, showed that eSyn-OPCs expressed more genes encoding postsynaptic proteins compared to aSyn-OPCs. (D-F) The mRNA levels of postsynaptic neurotransmitter receptor subunit genes for glutamate, GABA, and acetylcholine in eSyn-OPCs compared with other embryonic OPC clusters.

Article Snippet: To isolate eSyn-OPCs, PDGFRα + OPCs were passed on to a dish coated with GRB14 antibody (Proteintech, 15298-1-AP) for 20min at 37C.

Techniques:

Journal: bioRxiv

Article Title: Transcriptional Heterogeneity Reveals a Synaptic Gene Program in Developing and Adult Human Oligodendrocyte Precursor Cells

doi: 10.64898/2026.01.01.697017

Figure Lengend Snippet: (A) Identification of OPCs in the UMAP plot of brain cells obtained from a healthy 42-year-old male. OPCs were defined by co-expression of marker genes PDGFRA and OLIG2 , and mature oligodendrocytes were defined by co-expression of marker genes MAG , MOG , and MBP . (B) Re-clustering of the OPC cluster found in (A) based on differentially expressed genes. (C-F) Top pathways identified by GO analysis of differentially expressed genes in each OPC cluster from (B). A dult Cluster 4 (aC4) was identified as aSyn-OPCs.

Article Snippet: To isolate eSyn-OPCs, PDGFRα + OPCs were passed on to a dish coated with GRB14 antibody (Proteintech, 15298-1-AP) for 20min at 37C.

Techniques: Expressing, Marker

Journal: bioRxiv

Article Title: Transcriptional Heterogeneity Reveals a Synaptic Gene Program in Developing and Adult Human Oligodendrocyte Precursor Cells

doi: 10.64898/2026.01.01.697017

Figure Lengend Snippet: (A) An independent dataset from Schirmer et al. was used to identify OPCs from the single-nucleus RNA sequencing of cortical brain cells obtained from 6 healthy individuals of various ages (34 - 68 years old). OPCs were defined by co-expression of marker genes PDGFRA and OLIG2 , and mature oligodendrocytes were defined by co-expression of marker genes MAG , MOG , and MBP . (B) Re-clustering of the OPC cluster found in (A) based on differentially expressed genes. (C) Top seven pathways identified by GO analysis of differentially expressed genes in A dult Cluster 4 (aC4), identified as aSyn-OPCs.

Article Snippet: To isolate eSyn-OPCs, PDGFRα + OPCs were passed on to a dish coated with GRB14 antibody (Proteintech, 15298-1-AP) for 20min at 37C.

Techniques: RNA Sequencing, Expressing, Marker

Journal: bioRxiv

Article Title: Transcriptional Heterogeneity Reveals a Synaptic Gene Program in Developing and Adult Human Oligodendrocyte Precursor Cells

doi: 10.64898/2026.01.01.697017

Figure Lengend Snippet: (A) A representative image of a cortical section obtained from a PCW 22 sample. The location of the oSVZ region is indicated. (B) Immunohistochemistry with candidate eSyn-OPC markers. Images were taken from adjascent sections in the oSVZ region indicated in (A). Scale bar unit = um. (C) Corresponding mRNA expression levels of candidate markers used in the immunohistochemistry shown in (B). (D) A spatial transcriptomic sample from PCW 8 tissue. Negligible MT1E + spots (8 spot; 0.7%) were found despite the abundant PDGFRα + spots. (E) A spatial transcriptomic sample from PCW 19 tissue. (F) MT1E + spots were enriched in lower cortical layers whereas PDGFRα + spots were found across cortical layers. (G) The mRNA levels of neural progenitor ( EOMES ) and neural stem cell ( SOX2, ID4 ) markers in the MT1E + spots (containing eSyn-OPCs) compared to PDGFR α + / MT1E - spots (containing OPCs but not eSyn-OPCs) in PCW 19 sample.

Article Snippet: To isolate eSyn-OPCs, PDGFRα + OPCs were passed on to a dish coated with GRB14 antibody (Proteintech, 15298-1-AP) for 20min at 37C.

Techniques: Immunohistochemistry, Expressing

Journal: bioRxiv

Article Title: Transcriptional Heterogeneity Reveals a Synaptic Gene Program in Developing and Adult Human Oligodendrocyte Precursor Cells

doi: 10.64898/2026.01.01.697017

Figure Lengend Snippet: (A) Enrichment in cell surface marker genes in eSyn-OPCs compared to non-Syn-OPCs. GRB14 was identified as a unique marker. (B) A schematic illustrating the isolation of eSyn-OPCs based on double positivity for GRB14 and PDGFRα antibodies. (C) The isolated eSyn-OPCs are GRB14 + /PDGFRα + /OLIG2 + . Scale bar unit = um. (D) When isolated eSyn-OPCs were induced to differentiate with differentiation medium lacking growth factors, eSyn-OPCs differentiated into O4 + /MBP + /PLP1 + mature oligodendrocytes. Scale bar unit = um.

Article Snippet: To isolate eSyn-OPCs, PDGFRα + OPCs were passed on to a dish coated with GRB14 antibody (Proteintech, 15298-1-AP) for 20min at 37C.

Techniques: Marker, Isolation

Journal: Nature Communications

Article Title: TMEM120A maintains adipose tissue lipid homeostasis through ER CoA channeling

doi: 10.1038/s41467-025-67870-7

Figure Lengend Snippet: A mRNA levels of Tmem120a in various tissues. n = 3. B , C Protein expression of TMEM120A in various tissues. n = 4. D , E UMAP plot of total 18,542 nuclei isolated from gWAT of WT and Tmem120a AKO mice fed an NCD. Red color indicates higher expression levels, whereas grey color indicates lower expression levels. (Two replicates for the WT condition and one replicate for the Tmem120a AKO condition). Clusters are colored by cell types: Adipocyte, mesothelial cell (MC), lymphatic endothelial cell (LEC), vascular endothelial cell (VEC), adipocyte progenitor cell (APC), smooth muscle cell (SMC), macrophages, B cell and T cell. F mRNA levels of Tmem120a in floating adipocytes and F4/80 + macrophages and PDGFRα + progenitor cells from SVF isolated from iWAT and gWAT. n = 3. G Single-nucleus RNA analysis of adipocytes, APC, Trem2 + macrophages from gWAT of mice fed an NCD or HFD for 12 weeks (SRP426501). H Analysis of Tmem120a expression in gWAT of mice fed an NCD or HFD for 8 weeks, determined by publicly available transcriptomic data ( GSE182930 ). n = 3. I mRNA levels of Tmem120a in BAT, iWAT, and gWAT of WT mice fed an NCD or HFD for 8 weeks. n = 10. J Expression of TMEM120A across different cell types in human white adipose tissue, based on the public dataset GSE176171 . Adipose-derived stromal/stem progenitor cell (ASPC). mRNA levels ( K ) and correlation analysis ( L ) of TMEM120A gene expression in human subcutaneous adipose tissue of normal (BMI < 22 kg/m 2 ) and obese (BMI > 26 kg/m 2 ) patients. n = 12. M Pearson correlation between BMI and average TMEM120A expression in human adipocytes ( GSE176171 ). Each point represents an individual sample. The black dashed line indicates the linear regression fit, and the shaded area represents the 95% confidence interval. N Analysis of TMEM120A expression in human adipose tissue ((MHL): n = 14; (MHO): n = 25; (MUO): n = 27), determined by publicly available transcriptomic data ( GSE156906 ). Data are presented as mean values ± SEM. Each point represents a biological replicate. p values were determined by an unpaired two-sided Student’s t-test ( A, C , F – I , K , N ), or two-tailed Pearson correlation ( L , M ).

Article Snippet: The remaining F4/80 - cells were subjected to PDGFRα selection using a PDGFRα MicroBead kit (Miltenyi Biotec, cat#130-101-502).

Techniques: Expressing, Isolation, Derivative Assay, Gene Expression, Two Tailed Test

Journal: Journal of Neuroinflammation

Article Title: Remote ischemic postconditioning improves cognitive dysfunction after subarachnoid hemorrhage by driving metabolic reprogramming of border-associated macrophages through the IL-33/ST2 axis

doi: 10.1186/s12974-025-03587-0

Figure Lengend Snippet: Distribution patterns of BAMs after SAH and their effects on cognitive function and myelin. A Schematic diagram of the animal groups and experimental procedures. B Representative images of immunofluorescence staining for border-associated macrophages in the meninges of mice in the sham group and at 1 day, 7 days, and 14 days after SAH (white, Lyve1; red, CD206; blue, DAPI). C Representative images of immunofluorescence staining for border-associated macrophages in the perivascular space of the cortical region in mice from the sham group and at 1 day, 7 days, and 14 days after SAH (white, lectin; red, CD206; blue, DAPI). Scale bar, 100 μm. D Quantification of CD206 + BAMs in the meninges and perivascular space of the cortical region ( n = 6 per group; one-way ANOVA). E Representative images of myelin immunofluorescence staining in the SAH-Vehicle group and the SAH-CLO Lip group with border-associated macrophage depletion by clodronate liposomes (red, MBP; green, NF200; white, PDGFRα; blue, DAPI). Scale bar, 200 μm. F , G Quantitative analysis of MBP-positive (MBP +) myelin, NF200-positive (NF200 +) axons, and NF200 + /MBP + and MBP − /NF200. + areas in panel E ( n = 6 per group; T-test). H Statistical analysis of the trend in latency from day 1 to day 5 in the Morris water maze test, as well as the time spent in the target quadrant and the number of platform crossings ( n = 6 per group, two-way ANOVA). I Statistical chart of the time spent by the mice in the new arm and the percentage of alternations in the Y-maze test ( n = 6 per group, one-way ANOVA). J Statistical chart of the proportion of open arm entries to total arm entries by the mice in the elevated plus maze test ( n = 6 per group; one-way ANOVA)

Article Snippet: PDGFRα + OPCs were isolated from the mouse cortex at P2‒P4, trypsin-digested, filtered and sorted with PDGFRα + magnetic beads (Miltenyi Biotec, Germany, 130‒101‒502), inoculated into different vessels and cultured in medium containing specific growth factors.

Techniques: Immunofluorescence, Staining, Liposomes

Journal: Journal of Neuroinflammation

Article Title: Remote ischemic postconditioning improves cognitive dysfunction after subarachnoid hemorrhage by driving metabolic reprogramming of border-associated macrophages through the IL-33/ST2 axis

doi: 10.1186/s12974-025-03587-0

Figure Lengend Snippet: RIPostC treatment increases the abundance of BAMs after SAH and improves cognitive dysfunction. A Schematic diagram of the animal groups and experimental procedures. B , C Representative images of lower limb perfusion speckle in mice during the RIPostC treatment phase and statistical analysis of lower limb blood flow perfusion ROI values ( n = 6 per group; one-way ANOVA). D Representative images of meningeal border-associated macrophage immunofluorescence staining in the SAH group and SAH-RIPostC group (red, CD206; white, Lyve1; blue, DAPI). E Representative images of immunofluorescence staining of border-associated macrophages in the perivascular space of the cortical region in the SAH group and the SAH-RIPostC group (red, CD206; white, lectin; blue, DAPI). Scale bar, 100 μm. F Quantitative analysis of CD206 + BAMs in the meningeal and perivascular spaces of the cortical region, as shown in D-E ( n = 6 per group; T-test). G Representative images of myelin immunofluorescence staining in the SAH group and the SAH-RIPostC group (green, NF200; red, MBP; white, PDGFRα; blue, DAPI). Scale bar, 200 μm. H Quantitative analysis of NF200-positive (NF200 + ) axons, MBP-positive (MBP + ) myelin, and the MBP − /NF200 + and NF200 + /MBP + regions in G ( n = 6 per group; T-test). I Statistical analysis of the trend in latency from day 1 to day 5 in the Morris water maze test, as well as the time spent in the target quadrant and the number of platform crossings in both groups of mice ( n = 6 per group, two-way ANOVA and T-test). J Statistical chart of the time spent in the novel arm and the percentage of alternations in the Y-maze test ( n = 6 per group, T-test). K Statistical chart of the proportion of open arm entries to total arm entries in the elevated plus maze test ( n = 6 per group; T-test)

Article Snippet: PDGFRα + OPCs were isolated from the mouse cortex at P2‒P4, trypsin-digested, filtered and sorted with PDGFRα + magnetic beads (Miltenyi Biotec, Germany, 130‒101‒502), inoculated into different vessels and cultured in medium containing specific growth factors.

Techniques: Immunofluorescence, Staining

Journal: Journal of Neuroinflammation

Article Title: Remote ischemic postconditioning improves cognitive dysfunction after subarachnoid hemorrhage by driving metabolic reprogramming of border-associated macrophages through the IL-33/ST2 axis

doi: 10.1186/s12974-025-03587-0

Figure Lengend Snippet: Effects of BAMs from chimeric mice on OPC proliferation, migration, and myelination. A Primary cell grouping and experimental schematic diagram. BAMs were co-cultured with OPCs isolated via PDGFRα-positive magnetic beads in a Transwell chamber to evaluate OPC proliferation, migration, and differentiation. B Flow cytometry bar graph and statistical analysis of OPC proliferation after EdU-FITC incubation ( n = 3 per group, one-way ANOVA). C Representative images and quantification of migrating cells in the Transwell invasion assay used to assess OPC migration ability ( n = 6 per group; one-way ANOVA). D , E Representative images of cytoskeletal staining in OPCs co-cultured with BAMs for 1, 3, and 5 days (green, phalloidin-FITC; red, PDGFRα; blue, DAPI). F Representative images of Sholl analysis of OPC morphology (concentric circle interval = 50 μm). G - I Statistical graphs of the number of intersections (branching density) at different radii in OPCs co-cultured with BAMs for 1, 3, and 5 days. J , K Representative images of newly formed myelinating oligodendrocytes (OLs, MBP + ) induced by BAMs co-culture derived from OPCs after 1 day of T3 induction (green, Olig2; red, MBP; blue, DAPI), as well as statistical analysis of the percentage of MBP + /Olig2 + newly formed myelinating OLs ( n = 6 per group; one-way ANOVA)

Article Snippet: PDGFRα + OPCs were isolated from the mouse cortex at P2‒P4, trypsin-digested, filtered and sorted with PDGFRα + magnetic beads (Miltenyi Biotec, Germany, 130‒101‒502), inoculated into different vessels and cultured in medium containing specific growth factors.

Techniques: Migration, Cell Culture, Isolation, Magnetic Beads, Flow Cytometry, Incubation, Transwell Invasion Assay, Staining, Co-Culture Assay, Derivative Assay

Journal: Journal of Neuroinflammation

Article Title: Remote ischemic postconditioning improves cognitive dysfunction after subarachnoid hemorrhage by driving metabolic reprogramming of border-associated macrophages through the IL-33/ST2 axis

doi: 10.1186/s12974-025-03587-0

Figure Lengend Snippet: Evaluation of myelin injury in chimeric mice. A Animal grouping and experimental schematic diagram. B Representative immunofluorescence images of the corpus callosum region in the WT/WT RIPostC group and the WT/ST2 KO RIPostC group (green, MBP; blue, DAPI). C Statistical graph of the MBP + area (%) in the corpus callosum region ( n = 6 per group; T-test). D , E Representative images of CC1 and MCM2 immunofluorescence in the corpus callosum region in the WT/WT RIPostC group and the WT/ST2 KO RIPostC group (green, PDGFRα; red, CC1 or MCM2; blue, DAPI). F Statistical analysis of the number of CC1 + positive cells and the number of MCM2 + OPCs in the corpus callosum region ( n = 6 per group; T-test)

Article Snippet: PDGFRα + OPCs were isolated from the mouse cortex at P2‒P4, trypsin-digested, filtered and sorted with PDGFRα + magnetic beads (Miltenyi Biotec, Germany, 130‒101‒502), inoculated into different vessels and cultured in medium containing specific growth factors.

Techniques: Immunofluorescence

Journal: Journal of Neuroinflammation

Article Title: Remote ischemic postconditioning improves cognitive dysfunction after subarachnoid hemorrhage by driving metabolic reprogramming of border-associated macrophages through the IL-33/ST2 axis

doi: 10.1186/s12974-025-03587-0

Figure Lengend Snippet: Schematic diagram of this study. RIPostC treatment after SAH drives the IL33/ST2 axis, increases the abundance of BAMs in brain border tissue, induces amino acid metabolic reprogramming in BAMs, enhances OPCs proliferation, migration, and myelination, and improves cognitive dysfunction in mice after SAH

Article Snippet: PDGFRα + OPCs were isolated from the mouse cortex at P2‒P4, trypsin-digested, filtered and sorted with PDGFRα + magnetic beads (Miltenyi Biotec, Germany, 130‒101‒502), inoculated into different vessels and cultured in medium containing specific growth factors.

Techniques: Migration

Journal: Journal of Neuroinflammation

Article Title: Remote ischemic postconditioning improves cognitive dysfunction after subarachnoid hemorrhage by driving metabolic reprogramming of border-associated macrophages through the IL-33/ST2 axis

doi: 10.1186/s12974-025-03587-0

Figure Lengend Snippet: Effects of BAMs from chimeric mice on OPC proliferation, migration, and myelination. A Primary cell grouping and experimental schematic diagram. BAMs were co-cultured with OPCs isolated via PDGFRα-positive magnetic beads in a Transwell chamber to evaluate OPC proliferation, migration, and differentiation. B Flow cytometry bar graph and statistical analysis of OPC proliferation after EdU-FITC incubation ( n = 3 per group, one-way ANOVA). C Representative images and quantification of migrating cells in the Transwell invasion assay used to assess OPC migration ability ( n = 6 per group; one-way ANOVA). D , E Representative images of cytoskeletal staining in OPCs co-cultured with BAMs for 1, 3, and 5 days (green, phalloidin-FITC; red, PDGFRα; blue, DAPI). F Representative images of Sholl analysis of OPC morphology (concentric circle interval = 50 μm). G - I Statistical graphs of the number of intersections (branching density) at different radii in OPCs co-cultured with BAMs for 1, 3, and 5 days. J , K Representative images of newly formed myelinating oligodendrocytes (OLs, MBP + ) induced by BAMs co-culture derived from OPCs after 1 day of T3 induction (green, Olig2; red, MBP; blue, DAPI), as well as statistical analysis of the percentage of MBP + /Olig2 + newly formed myelinating OLs ( n = 6 per group; one-way ANOVA)

Article Snippet: PDGFRα + OPCs were isolated from the mouse cortex at P2‒P4, trypsin-digested, filtered and sorted with PDGFRα + magnetic beads (Miltenyi Biotec, Germany, 130‒101‒502), inoculated into different vessels and cultured in medium containing specific growth factors.

Techniques: Migration, Cell Culture, Isolation, Magnetic Beads, Flow Cytometry, Incubation, Transwell Invasion Assay, Staining, Co-Culture Assay, Derivative Assay