Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Resource
  • Published:

Transcriptional profiles of mouse oligodendrocyte precursor cells across the lifespan

Nature Aging volume 5, pages 675–690 (2025)Cite this article

Subjects

Abstract

Oligodendrocyte progenitor cells (OPCs) are highly dynamic, widely distributed glial cells of the central nervous system responsible for generating myelinating oligodendrocytes throughout life. However, the rates of OPC proliferation and differentiation decline dramatically with aging, which may impair homeostasis, remyelination and adaptive myelination during learning. To determine how aging influences OPCs, we generated a transgenic mouse line (Matn4-mEGFP) and performed single-cell RNA sequencing, providing enhanced resolution of transcriptional changes during key transitions from quiescence to proliferation and differentiation across the lifespan. We found that aging induces distinct transcriptomic changes in OPCs in different states, including enhanced activation of HIF-1α and WNT pathways. Pharmacological inhibition of these pathways in aged OPCs was sufficient to increase their ability to differentiate in vitro. Ultimately, Matn4-mEGFP mouse line and the sequencing dataset of cortical OPCs across ages will help to define the molecular changes guiding OPC behavior in various physiological and pathological contexts.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to the full article PDF.

USD 39.95

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Generation of an OPC reporter mouse line: Matn4-mEGFP.
Fig. 2: scRNA-seq analysis of mouse cortical OPCs across the lifespan.
Fig. 3: Different cycling OPC subtypes represent different stages of the cell cycle.
Fig. 4: Identification of a transitioning OPC population poised to undergo differentiation.
Fig. 5: Quiescent OPCs undergo aging-associated transcriptional changes.
Fig. 6: HIF-1α pathway is activated in aged OPCs, which functionally inhibits their differentiation.

Similar content being viewed by others

Data availability

All raw and preprocessed sequencing data generated for this study as well as the processed Monocle 3 cell_data_set (cds) object have been deposited in NCBI Gene Expression Omnibus (GEO) with accession code GSE249268. To promote open access to data, we deposited the annotated dataset onto the Chan Zuckerberg CELL by GENE Discover Platform (https://tinyurl.com/aging-opcs). Source data are provided with this paper. Any other data reported in this paper are available from the lead contact upon reasonable request.

Code availability

No new code for the analysis of data was created.

References

  1. Salas, A. et al. JAK-STAT pathway targeting for the treatment of inflammatory bowel disease. Nat. Rev. Gastroenterol. Hepatol. 17, 323–337 (2020).

    Article  PubMed  Google Scholar 

  2. Clarke, L. E. et al. Normal aging induces A1-like astrocyte reactivity. Proc. Natl Acad. Sci. USA 115, E1896–E1905 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Hammond, T. R. et al. Single-cell RNA sequencing of microglia throughout the mouse lifespan and in the injured brain reveals complex cell-state changes. Immunity 50, 253–271.e6 (2019).

    Article  CAS  PubMed  Google Scholar 

  4. Guttenplan, K. A. et al. Neurotoxic reactive astrocytes induce cell death via saturated lipids. Nature 599, 102–107 (2021).

    Article  CAS  PubMed  Google Scholar 

  5. Young, K. M. et al. Oligodendrocyte dynamics in the healthy adult CNS: evidence for myelin remodeling. Neuron 77, 873–885 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Hill, R. A., Li, A. M. & Grutzendler, J. Lifelong cortical myelin plasticity and age-related degeneration in the live mammalian brain. Nat. Neurosci. 21, 683–695 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Zonouzi, M. et al. Individual oligodendrocytes show bias for inhibitory axons in the neocortex. Cell Rep 27, 2799–2808.e3 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Bacmeister, C. M. et al. Motor learning promotes remyelination via new and surviving oligodendrocytes. Nat. Neurosci. 23, 819–831 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Hughes, E. G., Orthmann-Murphy, J. L., Langseth, A. J. & Bergles, D. E. Myelin remodeling through experience-dependent oligodendrogenesis in the adult somatosensory cortex. Nat. Neurosci. 21, 696–706 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Nishiyama, A., Shimizu, T., Sherafat, A. & Richardson, W. D. Life-long oligodendrocyte development and plasticity. Semin. Cell Dev. Biol. 116, 25–37 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Sim, F. J., Zhao, C., Penderis, J. & Franklin, R. J. M. The age-related decrease in CNS remyelination efficiency is attributable to an impairment of both oligodendrocyte progenitor recruitment and differentiation. J. Neurosci. 22, 2451–2459 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Neumann, B. et al. Metformin restores CNS remyelination capacity by rejuvenating aged stem cells. Cell Stem Cell 25, 473–485 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Kirby, L. et al. Oligodendrocyte precursor cells present antigen and are cytotoxic targets in inflammatory demyelination. Nat. Commun. 10, 3887 (2019).

    Article  PubMed  PubMed Central  Google Scholar 

  14. Falcão, A. M. et al. Disease-specific oligodendrocyte lineage cells arise in multiple sclerosis. Nat. Med. 24, 1837–1844 (2018).

    Article  PubMed  PubMed Central  Google Scholar 

  15. Park, H. et al. Single-cell RNA-sequencing identifies disease-associated oligodendrocytes in male APP NL-G-F and 5XFAD mice. Nat. Commun. 14, 802 (2023).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Dawson, M. R. L., Polito, A., Levine, J. M. & Reynolds, R. NG2-expressing glial progenitor cells: an abundant and widespread population of cycling cells in the adult rat CNS. Mol. Cell. Neurosci. 24, 476–488 (2003).

    Article  CAS  PubMed  Google Scholar 

  17. Uckelmann, H. et al. Extracellular matrix protein Matrilin-4 regulates stress-induced HSC proliferation via CXCR4. J. Exp. Med. 213, 1961–1971 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Hrvatin, S. et al. Single-cell analysis of experience-dependent transcriptomic states in the mouse visual cortex. Nat. Neurosci. 21, 120–129 (2018).

    Article  CAS  PubMed  Google Scholar 

  19. Nishiyama, A., Chang, A. & Trapp, B. D. NG2+ glial cells: a novel glial cell population in the adult brain. J. Neuropathol. Exp. Neurol. 58, 1113–1124 (1999).

    Article  CAS  PubMed  Google Scholar 

  20. Daneman, R. et al. The mouse blood-brain barrier transcriptome: a new resource for understanding the development and function of brain endothelial cells. PLoS ONE 5, e13741 (2010).

    Article  PubMed  PubMed Central  Google Scholar 

  21. Dani, N. et al. A cellular and spatial map of the choroid plexus across brain ventricles and ages. Cell 184, 3056–3074.e21 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Cembrowski, M. S., Wang, L., Sugino, K., Shields, B. C. & Spruston, N. Hipposeq: a comprehensive RNA-seq database of gene expression in hippocampal principal neurons. eLife 5, e14997 (2016).

    Article  PubMed  PubMed Central  Google Scholar 

  23. Sullivan, K. E. et al. Sharp cell-type-identity changes differentiate the retrosplenial cortex from the neocortex. Cell Rep. 42, 112206 (2023).

    Article  CAS  PubMed  Google Scholar 

  24. Hughes, E. G., Kang, S. H., Fukaya, M. & Bergles, D. E. Oligodendrocyte progenitors balance growth with self-repulsion to achieve homeostasis in the adult brain. Nat. Neurosci. 16, 668–676 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Ximerakis, M. et al. Single-cell transcriptomic profiling of the aging mouse brain. Nat. Neurosci. 22, 1696–1708 (2019).

    Article  CAS  PubMed  Google Scholar 

  26. Fard, M. K. et al. BCAS1 expression defines a population of early myelinating oligodendrocytes in multiple sclerosis lesions. Sci. Transl. Med. 9, eaam7816 (2017).

    Article  PubMed  PubMed Central  Google Scholar 

  27. Xiao, L. et al. Rapid production of new oligodendrocytes is required in the earliest stages of motor-skill learning. Nat. Neurosci. 19, 1210–1217 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. He, D. et al. lncRNA functional networks in oligodendrocytes reveal stage-specific myelination control by an lncOL1/Suz12 complex in the CNS. Neuron 93, 362–378 (2017).

    Article  CAS  PubMed  Google Scholar 

  29. Adams, K. L. et al. Endothelin-1 signaling maintains glial progenitor proliferation in the postnatal subventricular zone. Nat. Commun. 11, 2138 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Beiter, R. M. et al. Evidence for oligodendrocyte progenitor cell heterogeneity in the adult mouse brain. Sci. Rep. 12, 12921 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Psachoulia, K., Jamen, F., Young, K. M. & Richardson, W. D. Cell cycle dynamics of NG2 cells in the postnatal and ageing brain. Neuron Glia Biol. 5, 57–67 (2009).

    Article  PubMed  PubMed Central  Google Scholar 

  32. Kang, S. H., Fukaya, M., Yang, J. K., Rothstein, J. D. & Bergles, D. E. NG2+ CNS glial progenitors remain committed to the oligodendrocyte lineage in postnatal life and following neurodegeneration. Neuron 68, 668–681 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Spitzer, S. O. et al. Oligodendrocyte progenitor cells become regionally diverse and heterogeneous with age. Neuron 101, 459–471 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Siletti, K. et al. Transcriptomic diversity of cell types across the adult human brain. Science 382, eadd7046 (2023).

    Article  CAS  PubMed  Google Scholar 

  35. Stein-O’Brien, G. L. et al. Decomposing cell identity for transfer learning across cellular measurements, platforms, tissues, and species. Cell Syst 8, 395–411.e8 (2019).

    Article  PubMed  PubMed Central  Google Scholar 

  36. Sharma, G., Colantuoni, C., Goff, L. A., Fertig, E. J. & Stein-O’Brien, G. projectR: an R/Bioconductor package for transfer learning via PCA, NMF, correlation and clustering. Bioinformatics 36, 3592–3593 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. CZI Cell Science Program. CZ CELLxGENE Discover: a single-cell data platform for scalable exploration, analysis and modeling of aggregated data. Nucleic Acids Res. 53, D886–D900 (2021).

  38. Zheng, S. C. et al. Universal prediction of cell-cycle position using transfer learning. Genome Biol 23, 41 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Stein-O’Brien, G. L. et al. Enter the matrix: factorization uncovers knowledge from omics determining the dimensions of biology from omics data. Trends Genet. 34, 790–805 (2018).

    Article  PubMed  PubMed Central  Google Scholar 

  40. Baruch, K. et al. Aging. Aging-induced type I interferon signaling at the choroid plexus negatively affects brain function. Science 346, 89 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Xu, H., Dzhashiashvili, Y., Ming, G.-L., He, C. & Correspondence, B. P. m6A mRNA methylation is essential for oligodendrocyte maturation and CNS myelination. Neuron 105, 293–309 (2020).

    Article  CAS  PubMed  Google Scholar 

  42. Kotter, M. R., Stadelmann, C. & Hartung, H.-P. Enhancing remyelination in disease—can we wrap it up? Brain 134, 1882–1900 (2011).

    Article  PubMed  Google Scholar 

  43. Alto, L. T. & Terman, J. R. Semaphorins and their signaling mechanisms. Methods Mol. Biol. 1493, 1 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Cohen, R. I., Rottkamp, D. M., Maric, D., Barker, J. L. & Hudson, L. D. A role for semaphorins and neuropilins in oligodendrocyte guidance. J. Neurochem. 85, 1262–1278 (2003).

    Article  CAS  PubMed  Google Scholar 

  45. Bernard, F. et al. Role of transmembrane semaphorin Sema6A in oligodendrocyte differentiation and myelination. Glia 60, 1590–1604 (2012).

    Article  PubMed  Google Scholar 

  46. Segel, M. et al. Niche stiffness underlies the ageing of central nervous system progenitor cells. Nature 573, 130–134 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Emery, B. et al. Myelin gene regulatory factor is a critical transcriptional regulator required for CNS myelination. Cell 138, 172–185 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Zezula, J. et al. p21cip1 is required for the differentiation of oligodendrocytes independently of cell cycle withdrawal. EMBO Rep. 2, 27–34 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Qi, Y. et al. Control of oligodendrocyte differentiation by the Nkx2.2 homeodomain transcription factor. Development 128, 2723–2733 (2001).

    Article  CAS  PubMed  Google Scholar 

  50. Emery, B. & Lu, Q. R. Transcriptional and epigenetic regulation of oligodendrocyte development and myelination in the central nervous system. Cold Spring Harb. Perspect. Biol. 7, a020461 (2015).

    Article  PubMed  PubMed Central  Google Scholar 

  51. Iram, T. et al. Young CSF restores oligodendrogenesis and memory in aged mice via Fgf17. Nature 605, 509–515 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Zhou, Y. et al. Human and mouse single-nucleus transcriptomics reveal TREM2-dependent and -independent cellular responses in Alzheimer’s disease. Nat. Med. 26, 131 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Liu, J. Z. et al. BDH2 triggers ROS-induced cell death and autophagy by promoting Nrf2 ubiquitination in gastric cancer. J. Exp. Clin. Cancer Res. 39, 123 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Krämer, A., Green, J., Pollard, J. & Tugendreich, S. Causal analysis approaches in ingenuity pathway analysis. Bioinformatics 30, 523–530 (2014).

    Article  PubMed  Google Scholar 

  55. Neumann, B. et al. Myc determines the functional age state of oligodendrocyte progenitor cells. Nat Aging 1, 826–837 (2021).

    Article  PubMed  Google Scholar 

  56. Tang, K. et al. HIF-1α stimulates the progression of oesophageal squamous cell carcinoma by activating the Wnt/β-catenin signalling pathway. Br. J. Cancer 127, 474–487 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Xu, W. et al. Hypoxia activates Wnt/β-catenin signaling by regulating the expression of BCL9 in human hepatocellular carcinoma. Sci. Rep. 7, 40446 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Fancy, S. P. J. et al. Dysregulation of the Wnt pathway inhibits timely myelination and remyelination in the mammalian CNS. Genes Dev. 23, 1571 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. de la Fuente, A. G. et al. Changes in the oligodendrocyte progenitor cell proteome with ageing. Mol. Cell. Proteomics 19, 1281–1302 (2020).

    Article  PubMed  PubMed Central  Google Scholar 

  60. Zhang, P. et al. Senolytic therapy alleviates Aβ-associated oligodendrocyte progenitor cell senescence and cognitive deficits in an Alzheimer’s disease model. Nat. Neurosci. 22, 719–728 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  61. Chapman, T. W., Olveda, G. E., Bame, X., Pereira, E. & Hill, R. A. Oligodendrocyte death initiates synchronous remyelination to restore cortical myelin patterns in mice. Nat. Neurosci. 26, 555–569 (2023).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  62. von Streitberg, A. et al. NG2-glia transiently overcome their homeostatic network and contribute to wound closure after brain injury. Front Cell Dev Biol 9, 662056 (2021).

    Article  Google Scholar 

  63. Gibson, E. M. et al. Neuronal activity promotes oligodendrogenesis and adaptive myelination in the mammalian brain. Science 344, 1252304 (2014).

    Article  PubMed  PubMed Central  Google Scholar 

  64. Marques, S. et al. Oligodendrocyte heterogeneity in the mouse juvenile and adult central nervous system. Science 352, 1326–1329 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  65. Marques, S. et al. Transcriptional convergence of oligodendrocyte lineage progenitors during development. Dev. Cell 46, 504–517.e7 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  66. Kessaris, N. et al. Competing waves of oligodendrocytes in the forebrain and postnatal elimination of an embryonic lineage. Nat. Neurosci. 9, 173–179 (2006).

    Article  CAS  PubMed  Google Scholar 

  67. Tripathi, R. B. et al. Dorsally and ventrally derived oligodendrocytes have similar electrical properties but myelinate preferred tracts. J. Neurosci. 31, 6809–6819 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  68. Vallstedt, A., Klos, J. M. & Ericson, J. Multiple dorsoventral origins of oligodendrocyte generation in the spinal cord and hindbrain. Neuron 45, 55–67 (2005).

    Article  CAS  PubMed  Google Scholar 

  69. Crawford, A. H., Tripathi, R. B., Richardson, W. D. & Franklin, R. J. M. Developmental origin of oligodendrocyte lineage cells determines response to demyelination and susceptibility to age-associated functional decline. Cell Rep. 15, 761–773 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  70. Floriddia, E. M. et al. Distinct oligodendrocyte populations have spatial preference and different responses to spinal cord injury. Nat. Commun. 11, 5860 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  71. Hilscher, M. M. et al. Spatial and temporal heterogeneity in the lineage progression of fine oligodendrocyte subtypes. BMC Biol. 20, 122 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  72. Lin, J. P. et al. Transcriptomic architecture of nuclei in the marmoset CNS. Nat. Commun. 13, 5531 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  73. Hill, R. A. et al. NG2 cells in white matter but not gray matter proliferate in response to PDGF. J. Neurosci. 33, 14558–14566 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  74. Rivers, L. E. et al. PDGFRA/NG2 glia generate myelinating oligodendrocytes and piriform projection neurons in adult mice. Nat. Neurosci. 11, 1392–1401 (2008).

    Article  CAS  PubMed  Google Scholar 

  75. Gonsalvez, D. G. et al. The dynamics of oligodendrocyte generation: how distinct is the mouse from the human? Preprint at bioRxiv https://doi.org/10.1101/2019.12.23.887174 (2019).

  76. Wang, S. et al. Ermin is a p116 RIP -interacting protein promoting oligodendroglial differentiation and myelin maintenance. Glia 68, 2264–2276 (2020).

    Article  PubMed  Google Scholar 

  77. Xing, Y. L. et al. High-efficiency pharmacogenetic ablation of oligodendrocyte progenitor cells in the adult mouse CNS. Cell Rep. Methods 3, 100414 (2023).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  78. Buchanan, J. A. et al. Oligodendrocyte precursor cells ingest axons in the mouse neocortex. Proc. Natl Acad. Sci. USA 119, e2202580119 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  79. Auguste, Y. S. S. et al. Oligodendrocyte precursor cells engulf synapses during circuit remodeling in mice. Nat. Neurosci. 25, 1273–1278 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  80. Xiao, Y., Petrucco, L., Hoodless, L. J., Portugues, R. & Czopka, T. Oligodendrocyte precursor cells sculpt the visual system by regulating axonal remodeling. Nat. Neurosci. 25, 280–284 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  81. Levine, J. M. Increased expression of the NG2 chondroitin-sulfate proteoglycan after brain injury. J. Neurosci. 14, 4716–4730 (1994).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  82. Bradbury, E. J. et al. Chondroitinase ABC promotes functional recovery after spinal cord injury. Nature 416, 636–640 (2002).

    Article  CAS  PubMed  Google Scholar 

  83. Wei, H. et al. Glial progenitor heterogeneity and key regulators revealed by single-cell RNA sequencing provide insight to regeneration in spinal cord injury. Cell Rep 42, 112486 (2023).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  84. Schirmer, L. et al. Neuronal vulnerability and multilineage diversity in multiple sclerosis. Nature 573, 75–82 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  85. Jäkel, S. et al. Altered human oligodendrocyte heterogeneity in multiple sclerosis. Nature 566, 543–547 (2019).

    Article  PubMed  PubMed Central  Google Scholar 

  86. Safaiyan, S. et al. White matter aging drives microglial diversity. Neuron 109, 1100–1117.e10 (2021).

    Article  CAS  PubMed  Google Scholar 

  87. Kaya, T. et al. CD8+ T cells induce interferon-responsive oligodendrocytes and microglia in white matter aging. Nat. Neurosci. 25, 1446–1457 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  88. Harrington, E. P. et al. MHC class i and MHC class ii reporter mice enable analysis of immune oligodendroglia in mouse models of multiple sclerosis. eLife 12, e82938 (2023).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  89. Nicaise, A. M. et al. Cellular senescence in progenitor cells contributes to diminished remyelination potential in progressive multiple sclerosis. Proc. Natl Acad. Sci. USA 116, 9030–9039 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  90. Yuen, T. J. et al. Oligodendrocyte-encoded HIF function couples postnatal myelination and white matter angiogenesis. Cell 158, 383–396 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  91. Allan, K. C. et al. Non-canonical targets of HIF1a impair oligodendrocyte progenitor cell function. Cell Stem Cell 28, 257–272.e11 (2021).

    Article  CAS  PubMed  Google Scholar 

  92. Kuhlmann, T. et al. Differentiation block of oligodendroglial progenitor cells as a cause for remyelination failure in chronic multiple sclerosis. Brain 131, 1749–1758 (2008).

    Article  CAS  PubMed  Google Scholar 

  93. Call, C. L. & Bergles, D. E. Cortical neurons exhibit diverse myelination patterns that scale between mouse brain regions and regenerate after demyelination. Nat. Commun. 12, 4767 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  94. Melsted, P. et al. Modular, efficient and constant-memory single-cell RNA-seq preprocessing. Nat. Biotechnol. 39, 813–818 (2021).

    Article  CAS  PubMed  Google Scholar 

  95. Cao, J. et al. The single-cell transcriptional landscape of mammalian organogenesis. Nature 566, 496–502 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  96. Haghverdi, L., Lun, A. T. L., Morgan, M. D. & Marioni, J. C. Batch effects in single-cell RNA-sequencing data are corrected by matching mutual nearest neighbors. Nat. Biotechnol. 36, 421–427 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  97. Becht, E. et al. Dimensionality reduction for visualizing single-cell data using UMAP. Nat. Biotechnol. 37, 38–44 (2018).

    Article  Google Scholar 

  98. Schwarzkopf, M. et al. Hybridization chain reaction enables a unified approach to multiplexed, quantitative, high-resolution immunohistochemistry and in situ hybridization. Development 148, dev199847 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank C. Hawkins at JHMI Transgenic Core Laboratory for performing CRISPR–Cas9 microinjections and assisting in the generation of the Matn4-mEGFP mouse line. We also thank M. Pucak and A. Smirnov at JHMI Neuroscience Imaging Center for their assistance with image acquisition and analysis. R. Kawaguchi at UCLA provided oversight for the sequencing of 10x scRNA-seq libraries through AMRF. We also thank our colleagues for their invaluable support throughout this study. This research was supported by grants from the NIH (AG072305, NS041435), the Goldman Foundation and the Dr. Miriam and Sheldon G. Adelson Medical Research Foundation (AMRF). D.H. and A.K. were supported by fellowships from the NIH (F31NS110204 and F30AG084193, respectively). Y.M. was supported by a fellowship from the National MS Society (FG-1708-28962).

Author information

Author notes

  1. Dongeun Heo

    Present address: Vollum Institute, Oregon Health and Science University, Portland, OR, USA

  2. Björn Neumann & Robin J. M. Franklin

    Present address: Altos Labs - Cambridge Institute of Science, Granta Park, Cambridge, UK

Authors and Affiliations

  1. The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University, Baltimore, MD, USA

    Dongeun Heo, Anya A. Kim, Valerie N. Doze, Yu Kang T. Xu, Yevgeniya A. Mironova, Loyal A. Goff & Dwight E. Bergles

  2. Wellcome-Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, Cambridge, UK

    Björn Neumann & Robin J. M. Franklin

  3. Department of Genetic Medicine, Johns Hopkins School of Medicine, Baltimore, MD, USA

    Jared Slosberg & Loyal A. Goff

  4. Kavli Neuroscience Discovery Institute, Johns Hopkins University, Baltimore, MD, USA

    Jared Slosberg, Loyal A. Goff & Dwight E. Bergles

Authors
  1. Dongeun Heo
    View author publications

    Search author on:PubMed Google Scholar

  2. Anya A. Kim
    View author publications

    Search author on:PubMed Google Scholar

  3. Björn Neumann
    View author publications

    Search author on:PubMed Google Scholar

  4. Valerie N. Doze
    View author publications

    Search author on:PubMed Google Scholar

  5. Yu Kang T. Xu
    View author publications

    Search author on:PubMed Google Scholar

  6. Yevgeniya A. Mironova
    View author publications

    Search author on:PubMed Google Scholar

  7. Jared Slosberg
    View author publications

    Search author on:PubMed Google Scholar

  8. Loyal A. Goff
    View author publications

    Search author on:PubMed Google Scholar

  9. Robin J. M. Franklin
    View author publications

    Search author on:PubMed Google Scholar

  10. Dwight E. Bergles
    View author publications

    Search author on:PubMed Google Scholar

Contributions

D.H., L.A.G. and D.E.B. conceived the project, designed the experiments, and wrote the manuscript with input from the other authors. A.A.K. and V.N.D. performed FISH validation experiments. B.N. and R.J.M.F. performed and analyzed in vitro pharmacological validation experiments. Y.K.T.X. and Y.A.M. performed in vivo imaging of OPC reporter mouse lines. D.H., J.S. and L.A.G. performed bioinformatical analyses of the scRNA-seq dataset. L.A.G., R.J.M.F. and D.E.B. provided funding for the study.

Corresponding author

Correspondence to Dwight E. Bergles.

Ethics declarations

Competing interests

The authors declare no competing interests.

Peer review

Peer review information

Nature Aging thanks the anonymous reviewer(s) for their contribution to the peer review of this work.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Extended Data Fig. 1 Matn4-mEGFP expression is restricted to OPCs and a subset of neurons.

a. Matn4 expression is specific to OPCs and newly-formed oligodendrocytes (NFO) in 6-7 week-old mouse V1 cortex (reanalysis of a publicly available scRNA-seq dataset18). b. Genotyping result of Matn4-mEGFP mouse line. Wildtype (wt) band size is 178 bp whereas the mutant, knock-in band size is 356 bp. c. EGFP signal in the optic nerve of Matn4-mEGFP mouse line is restricted to NG2 + PDGFRα+ OPCs. d. Matn4-mEGFP is also expressed by hippocampal granule cells and neurons in the somatosensory cortex barrel field and retrosplenial cortex. e. Matn4-mEGFP signal is absent from Iba1+ microglia and GFAP+ astrocytes. f. In vivo imaging of GFP+ cells in Matn4-mEGFP, NG2-mEGFP, and Pdgfra-CreER; RCE mouse lines. None of the vascular cells (red arrowheads) express EGFP in the cortex of Matn4-mEGFP mice.

Extended Data Fig. 2 Preprocessing of the OPC scRNA-seq dataset.

a. Expression of oligodendrocyte lineage cell genes in the uncleaned dataset. Most cells in the dataset express Cspg4, Pdgfra, and Olig2 (OPCs) or Enpp6 and Olig2 (differentiating OPCs). b. Only a small group of cells that were FACS isolated from Matn4-mEGFP mouse line express non-oligodendrocyte lineage cell genes. c. UMAP plot of uncleaned dataset colorized by the percentage of mitochondrial-related genes (cutoff at 10%). Those cells with relatively high mitochondrial gene ratio (>5%) were removed for downstream analyses. d. Expression of classic oligodendrocyte lineage marker genes (oligodendrocyte lineage: Olig2, Sox10; OPC: Pdgfra, Cspg4, Matn4; differentiating OPC: Bcas1, Enpp6, 9630013A20Rik; oligodendrocyte: Mbp, Mobp) as well as the subtype marker genes identified in this study (Cycling OPC: Top2a, Mcm3, Mki67; Transitioning OPC: Gap43, Rplp0) in the cleaned, preprocessed, final dataset.

Extended Data Fig. 3 Fluorescent in situ hybridization (FISH) for Cycling and Differentiating OPC subtypes.

a. FISH for Top2a, Pdgfra, and Sox10 to identify Cycling OPC in situ in postnatal day 9 (P9) mouse brain. b. Comparison of the density of Cycling OPC in highly myelinated, somatosensory cortex (SS) and that in sparsely myelinated, temporal association cortex (TEA). c. Quantification of the density of Cycling OPC (Top2a+ Pdgfra + ) in SS and TEA. d. FISH for LncOL1 to identify Differentiating OPC in situ in P74 mouse brain. e. Quantification of the frequency of LncOL1+ Differentiating OPC in SS and TEA. The exact p-values are reported in Source Data file for Extended Data Fig. 3.

Source data

Extended Data Fig. 4 OPC subtype-specific gene patterns are shared between mouse and human cortical OPCs.

a. UMAP plot of the dataset colorized by their four age groups (blue: 29-yr old, red: 42-yr old, green: 50-yr old, and purple: 60-yr old). b. UMAP plot of 26,357 human cortical OPCs colorized by their identified subtypes (quiescent OPC, cycling OPC, differentiating OPC, and oligodendrocytes). c. Expression of classic oligodendrocyte lineage marker genes (oligodendrocyte lineage: OLIG2, SOX10; OPC: PDGFRA, CSPG4, MATN4; differentiating OPC: BCAS1, ENPP6; oligodendrocyte: MBP, MOBP) as well as the subtype marker genes identified in this study (Cycling OPC: TOP2A, MCM3, MKI67; Transitioning OPC: GAP43, RPLP0) in the human cortical OPC dataset. d. UMAP plots of mouse Cycling OPC gene pattern 5 projected on the mouse scRNA-seq dataset and on the human snRNA-seq dataset. e. UMAP plots of mouse Differentiating OPC gene pattern 25 projected on the mouse scRNA-seq dataset and on the human snRNA-seq dataset. f. Dot plot of HIF1A and CTNNB1 expression in quiescent OPCs in the human cortex across aging.

Extended Data Fig. 5 Expression changes in individual cycling genes and groups of genes in Cycling OPC 1 and directly anteceding Quiescent OPC.

a. Expression levels of known cycling genes enriched in cycling OPCs are comparable in Cycling OPC 1 throughout aging. b. NMF gene patterns that are associated with either aged (P180-720) or young (P30) OPCs.

Extended Data Fig. 6 OPCs upregulate C4b, Hif1a, and Ctnnb1 mRNA with aging.

a. FISH with immunofluorescence staining (IF) for Pdgfra and C4b in P35 and P315 Matn4-mEGFP mouse cortex. OPC cell body masks were created based on EGFP fluorescence and Pdgfra FISH signal and used to quantify C4b transcript puncta/OPC (Two-tailed, Mann-Whitney U test, n = 65, 49 cells, * p-value = 0.0177, error bars = SEM). b. FISH with IF staining for Pdgfra and Hif1a in the P35 and P315 Matn4-mEGFP mouse cortex. Hif1a transcript puncta/OPC was quantified as described above (Two-tailed, Mann-Whitney U test, n = 56, 71 cells, * p-value = 0.0385, error bars = SEM). c. FISH with IF staining for Pdgfra and Ctnnb1 in the P35 and P315 Matn4-mEGFP mouse cortex (Two-tailed, Mann-Whitney U test, n = 59, 55 cells, ** p-value = 0.0069, error bars = SEM). The exact p-values are reported in Source Data file for Extended Data Fig. 6.

Source data

Extended Data Fig. 7 Wnt signaling pathway is activated in aged OPCs and may contribute to their decreased differentiation potential.

a. Dot plot of Ctnnb1 expression in Quiescent OPC from P180, P360, and P720 timepoints. b. Schematic of how two different Wnt inhibitors (IWP-2 and XAV939) differentially block Wnt signaling pathway. IWP-2 globally inhibits the Wnt pathway whereas XAV939 preferentially inhibits the canonical Wnt signaling pathway. Both non-canonical and canonical Wnt signaling pathways have been shown to regulate DNA damage response. c. Quantification of MBP+ differentiating OPC/Olig2+ oligodendrocyte proportions with and without Wnt inhibitor treatments in OPCs isolated from YA or AA (two-way ANOVA with Tukey’s multiple comparisons test, n = 6, 6, * p-value < 0.05, ** p-value < 0.01, error bars = SEM). The exact p-values are reported in Source Data file for Extended Data Fig. 7.

Source data

Extended Data Fig. 8 Matn4-mEGFP signal is restricted to OPCs even after a stab wound injury.

IHC against NG2 (red) and EGFP (green) was performed on the Matn4-mEGFP mouse following a stab wound injury to demonstrate the utility of the mouse line in studying OPC dynamics following injury and inflammation.

Supplementary information

Supplementary Information (download PDF )

Supplementary Figs. 1 and 2.

Reporting Summary (download PDF )

Supplementary Tables 1–5. (download XLSX )

Table 1, List of marker genes used to define OPC subtypes. Table 2, List of genes corresponding to different gene modules associated with Quiescent, Transitioning and Differentiating OPC clusters. Table 3, Pseudotime differential gene expression (graph_test) of genes that encode transcription factors along the OPC differentiation trajectory. Table 4, Differential gene expression results of Quiescent OPCs in aging (P30 versus P180, P360 and P720). Table 5, Gene weights for Cycling and Differentiating OPC subtypes.

Supplementary Video 1 (download AVI )

1-h time-lapse imaging of OPCs in the motor cortex of Matn4-mEGFP.

Supplementary Video 2 (download AVI )

Z-stack image of the OPCs pseudocolored in green (at baseline of imaging) and magenta (50 min after baseline).

Source data

Source Data Fig. 1 (download XLSX )

Quantification for the graphs Fig. 1f–g.

Source Data Fig. 2 (download XLSX )

Raw cell counts for Fig. 2e.

Source Data Fig. 6 (download XLSX )

Quantification for the graphs Fig. 6c,e.

Source Data Extended Data Fig. 3 (download XLSX )

Quantification for the graphs Extended Data Fig. 3c,e.

Source Data Extended Data Fig. 6 (download XLSX )

Raw puncta quantification for the graphs Extended Data Fig. 6.

Source Data Extended Data Fig. 7 (download XLSX )

Quantification for Fig. 7c.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Heo, D., Kim, A.A., Neumann, B. et al. Transcriptional profiles of mouse oligodendrocyte precursor cells across the lifespan. Nat Aging 5, 675–690 (2025). https://doi.org/10.1038/s43587-025-00840-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Version of record:

  • Issue date:

  • DOI: https://doi.org/10.1038/s43587-025-00840-2

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing