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