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In vivo reprogramming of astrocytes to neuroblasts in the adult brain

Nature Cell Biology volume 15, pages 11641175 (2013) | Download Citation

Abstract

Adult differentiated cells can be reprogrammed into pluripotent stem cells or lineage-restricted proliferating precursors in culture; however, this has not been demonstrated in vivo. Here, we show that the single transcription factor SOX2 is sufficient to reprogram resident astrocytes into proliferative neuroblasts in the adult mouse brain. These induced adult neuroblasts (iANBs) persist for months and can be generated even in aged brains. When supplied with BDNF and noggin or when the mice are treated with a histone deacetylase inhibitor, iANBs develop into electrophysiologically mature neurons, which functionally integrate into the local neural network. Our results demonstrate that adult astrocytes exhibit remarkable plasticity in vivo, a feature that might have important implications in regeneration of the central nervous system using endogenous patient-specific glial cells.

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References

  1. 1.

    , & Mechanisms and functional implications of adult neurogenesis. Cell 132, 645–660 (2008).

  2. 2.

    & The glial nature of embryonic and adult neural stem cells. Annu. Rev. Neurosci. 32, 149–184 (2009).

  3. 3.

    , , & Adult neural stem cells in the mammalian central nervous system. Cell Res. 19, 672–682 (2009).

  4. 4.

    Orchestrating transcriptional control of adult neurogenesis. Genes Dev. 26, 1010–1021 (2012).

  5. 5.

    , , & Milestones of neuronal development in the adult hippocampus. Trends Neurosci. 27, 447–452 (2004).

  6. 6.

    et al. Homeostatic neurogenesis in the adult hippocampus does not involve amplification of Ascl1(high) intermediate progenitors. Nat. Commun. 3, 670 (2012).

  7. 7.

    , , , & Neuronal replacement from endogenous precursors in the adult brain after stroke. Nat. Med. 8, 963–970 (2002).

  8. 8.

    , & Induction of neurogenesis in the neocortex of adult mice. Nature 405, 951–955 (2000).

  9. 9.

    et al. Chordin-induced lineage plasticity of adult SVZ neuroblasts after demyelination. Nat. Neurosci. 13, 541–550 (2010).

  10. 10.

    & Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126, 663–676 (2006).

  11. 11.

    et al. Direct reprogramming of fibroblasts into neural stem cells by defined factors. Cell Stem Cell 10, 465–472 (2012).

  12. 12.

    et al. Direct reprogramming of mouse fibroblasts to neural progenitors. Proc. Natl Acad. Sci. USA 108, 7838–7843 (2011).

  13. 13.

    et al. Direct reprogramming of mouse and human fibroblasts into multipotent neural stem cells with a single factor. Cell Stem Cell 11, 100–109 (2012).

  14. 14.

    et al. Direct conversion of fibroblasts into stably expandable neural stem cells. Cell Stem Cell 10, 473–479 (2012).

  15. 15.

    , , , & Direct conversion of mouse fibroblasts to self-renewing, tripotent neural precursor cells. Proc. Natl Acad. Sci. USA 109, 2527–2532 (2012).

  16. 16.

    et al. Direct reprogramming of adult human fibroblasts to functional neurons under defined conditions. Cell Stem Cell 9, 113–118 (2011).

  17. 17.

    et al. Direct generation of functional dopaminergic neurons from mouse and human fibroblasts. Nature 476, 224–227 (2011).

  18. 18.

    et al. Directing astroglia from the cerebral cortex into subtype specific functional neurons. PLoS Biol. 8, e1000373 (2010).

  19. 19.

    et al. Glial cells generate neurons: the role of the transcription factor Pax6. Nat. Neurosci. 5, 308–315 (2002).

  20. 20.

    et al. Direct conversion of human fibroblasts to dopaminergic neurons. Proc. Natl Acad. Sci. USA 108, 10343–10348 (2011).

  21. 21.

    et al. Direct conversion of fibroblasts to functional neurons by defined factors. Nature 463, 1035–1041 (2010).

  22. 22.

    et al. Reprogramming of pericyte-derived cells of the adult human brain into induced neuronal cells. Cell Stem Cell 11, 471–476 (2012).

  23. 23.

    , , , & In vivo reprogramming of adult pancreatic exocrine cells to beta-cells. Nature 455, 627–632 (2008).

  24. 24.

    et al. Heart repair by reprogramming non-myocytes with cardiac transcription factors. Nature 485, 599–604 (2012).

  25. 25.

    et al. In vivo reprogramming of murine cardiac fibroblasts into induced cardiomyocytes. Nature 485, 593–598 (2012).

  26. 26.

    et al. Generation of induced neurons via direct conversion in vivo. Proc. Natl Acad. Sci. USA 110, 7038–7043 (2013).

  27. 27.

    , & The stem cell potential of glia: lessons from reactive gliosis. Nat. Rev. Neurosci. 12, 88–104 (2011).

  28. 28.

    & Astrocytes: biology and pathology. Acta Neuropathol. 119, 7–35 (2010).

  29. 29.

    Molecular dissection of reactive astrogliosis and glial scar formation. Trends Neurosci. 32, 638–647 (2009).

  30. 30.

    et al. Direct reprogramming of human astrocytes into neural stem cells and neurons. Exp. Cell Res. 318, 1528–1541 (2012).

  31. 31.

    , , & GFAP promoter elements required for region-specific and astrocyte-specific expression. Glia 56, 481–493 (2008).

  32. 32.

    et al. SOX9 induces and maintains neural stem cells. Nat. Neurosci. 13, 1181–1189 (2010).

  33. 33.

    et al. Postnatal deletion of Numb/Numblike reveals repair and remodeling capacity in the subventricular neurogenic niche. Cell 127, 1253–1264 (2006).

  34. 34.

    et al. hGFAP-cre transgenic mice for manipulation of glial and neuronal function in vivo. Genesis 31, 85–94 (2001).

  35. 35.

    et al. Pten deletion in adult neural stem/progenitor cells enhances constitutive neurogenesis. J. Neurosci. 29, 1874–1886 (2009).

  36. 36.

    et al. Molecular diversity of astrocytes with implications for neurological disorders. Proc. Natl Acad. Sci. USA 101, 8384–8389 (2004).

  37. 37.

    , , , & Involvement of dopaminergic neuronal cystatin C in neuronal injury-induced microglial activation and neurotoxicity. J. Neurochem. 122, 752–763 (2012).

  38. 38.

    et al. Cystatin C, a cysteine protease inhibitor, is persistently up-regulated in neurons and glia in a rat model for mesial temporal lobe epilepsy. Eur. J. Neurosci. 14, 1485–1491 (2001).

  39. 39.

    et al. Amyotrophic lateral sclerosis with dementia: an autopsy case showing many Bunina bodies, tau-positive neuronal and astrocytic plaque-like pathologies, and pallido-nigral degeneration. Acta Neuropathol. 112, 633–645 (2006).

  40. 40.

    , , & Physiology of microglia. Physiol. Rev. 91, 461–553 (2011).

  41. 41.

    , & Heterogeneity of astrocyte resting membrane potentials and intercellular coupling revealed by whole-cell and gramicidin-perforated patch recordings from cultured neocortical and hippocampal slice astrocytes. J. Neurosci. 17, 6850–6863 (1997).

  42. 42.

    , , , & Local generation of glia is a major astrocyte source in postnatal cortex. Nature 484, 376–380 (2012).

  43. 43.

    , , & NG2-glia as multipotent neural stem cells: fact or fantasy? Neuron 70, 661–673 (2011).

  44. 44.

    , & NG2 cells generate both oligodendrocytes and gray matter astrocytes. Development 135, 145–157 (2008).

  45. 45.

    , & Temporally controlled targeted somatic mutagenesis in the mouse brain. Eur. J. Neurosci. 14, 1777–1783 (2001).

  46. 46.

    et al. Cyclin-dependent kinase 5 governs learning and synaptic plasticity via control of NMDAR degradation. Nat. Neurosci. 10, 880–886 (2007).

  47. 47.

    et al. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat. Neurosci. 13, 133–140 (2010).

  48. 48.

    , , & Adenovirally expressed noggin and brain-derived neurotrophic factor cooperate to induce new medium spiny neurons from resident progenitor cells in the adult striatal ventricular zone. J. Neurosci. 24, 2133–2142 (2004).

  49. 49.

    et al. Induction of neostriatal neurogenesis slows disease progression in a transgenic murine model of Huntington disease. J. Clin. Invest. 117, 2889–2902 (2007).

  50. 50.

    , , , & Histone deacetylase inhibition-mediated neuronal differentiation of multipotent adult neural progenitor cells. Proc. Natl Acad. Sci. USA 101, 16659–16664 (2004).

  51. 51.

    et al. Mood stabilizer valproate promotes ERK pathway-dependent cortical neuronal growth and neurogenesis. J. Neurosci. 24, 6590–6599 (2004).

  52. 52.

    Physiology and pharmacology of striatal neurons. Annu. Rev. Neurosci. 32, 127–147 (2009).

  53. 53.

    , , & Heterogeneity and diversity of striatal GABAergic interneurons. Front. Neuroanat. 4, 150 (2010).

  54. 54.

    & The Yin and Yang of Sox proteins: Activation and repression in development and disease. J. Neurosci. Res. 87, 3277–3287 (2009).

  55. 55.

    , , & SOX2 functions to maintain neural progenitor identity. Neuron 39, 749–765 (2003).

  56. 56.

    et al. Sox2 deficiency causes neurodegeneration and impaired neurogenesis in the adult mouse brain. Development 131, 3805–3819 (2004).

  57. 57.

    et al. Consequence of the loss of Sox2 in the developing brain of the mouse. FEBS Lett. 582, 2811–2815 (2008).

  58. 58.

    et al. Hippocampal development and neural stem cell maintenance require Sox2-dependent regulation of Shh. Nat. Neurosci. 12, 1248–1256 (2009).

  59. 59.

    et al. Role of Sox2 in the development of the mouse neocortex. Dev. Biol. 295, 52–66 (2006).

  60. 60.

    & Neurotrophins: roles in neuronal development and function. Annu. Rev. Neurosci. 24, 677–736 (2001).

  61. 61.

    et al. Anterograde transport of brain-derived neurotrophic factor and its role in the brain. Nature 389, 856–860 (1997).

  62. 62.

    et al. Noggin antagonizes BMP signaling to create a niche for adult neurogenesis. Neuron 28, 713–726 (2000).

  63. 63.

    , , , & Chronic lithium treatment increases the expression of brain-derived neurotrophic factor in the rat brain. Psychopharmacology (Berl) 158, 100–106 (2001).

  64. 64.

    et al. Valproate regulates GSK-3-mediated axonal remodeling and synapsin I clustering in developing neurons. Mol. Cell Neurosci. 20, 257–270 (2002).

  65. 65.

    et al. Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev. Biol. 1, 4 (2001).

  66. 66.

    , , & Activation of postnatal neural stem cells requires nuclear receptor TLX. J. Neurosci. 31, 13816–13828 (2011).

  67. 67.

    et al. Sox3 expression identifies neural progenitors in persistent neonatal and adult mouse forebrain germinative zones. J. Comp. Neurol. 497, 88–100 (2006).

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Acknowledgements

We thank members of the Zhang laboratory for discussions and reagents. We also thank J. Bibb (UT Southwestern, USA) and P. Chambon (I.G.B.M.C., France) for providing PrP–CreERT mice, J. Hsieh (UT Southwestern, USA), C. Kuo (Duke University, USA) and Y. Jan (UCSF, USA) for NesCreERTM mice, M. Klymkowsky (UC Boulder, USA) for SOX3 antibody, K. Huber for sharing equipment, and E. Olson for critical reading of the manuscript. C-L.Z. is a W. W. Caruth, Jr. Scholar in Biomedical Research. This work was supported by The American Heart Association (09SDG2260602), The Whitehall Foundation Award (2009-12-05), The Welch Foundation Award (I-1724), The Ellison Medical Foundation Award (AG-NS-0753-11), and NIH Grants (1DP2OD006484 and R01NS070981; to C-L.Z.).

Author information

Affiliations

  1. Department of Molecular Biology, University of Texas Southwestern Medical Center, 6000 Harry Hines Boulevard, Dallas, Texas 75390, USA

    • Wenze Niu
    • , Tong Zang
    • , Yuhua Zou
    • , Sanhua Fang
    • , Derek K. Smith
    •  & Chun-Li Zhang
  2. Department of Pharmacology and Institute of Neuroscience, School of Medicine, Zhejiang University, 866 Yuhangtang Road, Hangzhou, Zhejiang 310058, China

    • Sanhua Fang
  3. Department of Neurology, University of Texas Southwestern Medical Center, 6000 Harry Hines Boulevard, Dallas, Texas 75390, USA

    • Robert Bachoo

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Contributions

W.N., T.Z. and C-L.Z. conceived and designed the experiments. W.N., T.Z., Y.Z. and C-L.Z. performed experiments. S.F. contributed to partial surgical experiments. D.K.S. critically reviewed and edited the manuscript. R.B. developed the Cst3–CreERT2 transgenic mice. W.N., T.Z. and C-L.Z. analysed data and prepared the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Chun-Li Zhang.

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https://doi.org/10.1038/ncb2843

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