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Forebrain ependymal cells are Notch-dependent and generate neuroblasts and astrocytes after stroke

Nature Neuroscience volume 12, pages 259–267 (2009)Cite this article

Abstract

Neurons are continuously generated from stem cells in discrete regions in the adult mammalian brain. We found that ependymal cells lining the lateral ventricles were quiescent and did not contribute to adult neurogenesis under normal conditions in mice but instead gave rise to neuroblasts and astrocytes in response to stroke. Ependymal cell quiescence was actively maintained by canonical Notch signaling. Inhibition of this pathway in uninjured animals allowed ependymal cells to enter the cell cycle and produce olfactory bulb neurons, whereas forced Notch signaling was sufficient to block the ependymal cell response to stroke. Ependymal cells were depleted by stroke and failed to self-renew sufficiently to maintain their own population. Thus, although ependymal cells act as primary cells in the neural lineage to produce neurons and glial cells after stroke, they do not fulfill defining criteria for stem cells under these conditions and instead serve as a reservoir that is recruited by injury.

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Figure 1: Ependymal cells generate neuroblasts and astrocytes in response to stroke.
Figure 2: Ependymal cell loss after stroke is inhibited by constitutive Notch signaling.
Figure 3: The Notch signaling pathway in the adult lateral ventricle wall stem cell niche.
Figure 4: Canonical Notch signaling maintains ependymal phenotype.
Figure 5: RBP-J is required for ependymal cell maintenance.
Figure 6: Proliferation of ependymal cell progeny in the absence of RBP-J.
Figure 7: Ependymal cells generate neuroblasts after RBP-J deletion.
Figure 8: Ependymal cells give rise to olfactory bulb neurons after RBP-J deletion.

References

  1. Mirzadeh, Z., Merkle, F.T., Soriano-Navarro, M., Garcia-Verdugo, J.M. & Alvarez-Buylla, A. Neural stem cells confer unique pinwheel architecture to the ventricular surface in neurogenic regions of the adult brain. Cell Stem Cell 3, 265–278 (2008).

    Article  CAS  Google Scholar 

  2. Doetsch, F., Caille, I., Lim, D.A., Garcia-Verdugo, J.M. & Alvarez-Buylla, A. Subventricular zone astrocytes are neural stem cells in the adult mammalian brain. Cell 97, 703–716 (1999).

    Article  CAS  Google Scholar 

  3. Johansson, C.B. et al. Identification of a neural stem cell in the adult mammalian central nervous system. Cell 96, 25–34 (1999).

    Article  CAS  Google Scholar 

  4. Meletis, K. et al. Spinal cord injury reveals multilineage differentiation of ependymal cells. PLoS Biol. 6, e182 (2008).

    Article  Google Scholar 

  5. Weigmann, A., Corbeil, D., Hellwig, A. & Huttner, W.B. Prominin, a novel microvilli-specific polytopic membrane protein of the apical surface of epithelial cells, is targeted to plasmalemmal protrusions of non-epithelial cells. Proc. Natl. Acad. Sci. USA 94, 12425–12430 (1997).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  7. Sakakibara, S. et al. RNA-binding protein Musashi family: roles for CNS stem cells and a subpopulation of ependymal cells revealed by targeted disruption and antisense ablation. Proc. Natl. Acad. Sci. USA 99, 15194–15199 (2002).

    Article  CAS  Google Scholar 

  8. Doetsch, F., Garcia-Verdugo, J.M. & Alvarez-Buylla, A. Cellular composition and three-dimensional organization of the subventricular germinal zone in the adult mammalian brain. J. Neurosci. 17, 5046–5061 (1997).

    Article  CAS  Google Scholar 

  9. Ellis, P. et al. SOX2, a persistent marker for multipotential neural stem cells derived from embryonic stem cells, the embryo or the adult. Dev. Neurosci. 26, 148–165 (2004).

    Article  CAS  Google Scholar 

  10. Götz, M. & Huttner, W.B. The cell biology of neurogenesis. Nat. Rev. Mol. Cell Biol. 6, 777–788 (2005).

    Article  Google Scholar 

  11. Spassky, N. et al. Adult ependymal cells are postmitotic and are derived from radial glial cells during embryogenesis. J. Neurosci. 25, 10–18 (2005).

    Article  CAS  Google Scholar 

  12. Gregg, C. & Weiss, S. Generation of functional radial glial cells by embryonic and adult forebrain neural stem cells. J. Neurosci. 23, 11587–11601 (2003).

    Article  CAS  Google Scholar 

  13. Taylor, M.D. et al. Radial glia cells are candidate stem cells of ependymoma. Cancer Cell 8, 323–335 (2005).

    Article  CAS  Google Scholar 

  14. Zhang, R.L. et al. Stroke induces ependymal cell transformation into radial glia in the subventricular zone of the adult rodent brain. J. Cereb. Blood Flow. Metab. 27, 1201–1212 (2007).

    Article  Google Scholar 

  15. Coskun, V. et al. CD133+ neural stem cells in the ependyma of mammalian postnatal forebrain. Proc. Natl. Acad. Sci. USA 105, 1026–1031 (2008).

    Article  CAS  Google Scholar 

  16. Capela, A. & Temple, S. LeX/ssea-1 is expressed by adult mouse CNS stem cells, identifying them as nonependymal. Neuron 35, 865–875 (2002).

    Article  Google Scholar 

  17. Louvi, A. & Artavanis-Tsakonas, S. Notch signaling in vertebrate neural development. Nat. Rev. Neurosci. 7, 93–102 (2006).

    Article  CAS  Google Scholar 

  18. Gaiano, N., Nye, J.S. & Fishell, G. Radial glial identity is promoted by Notch1 signaling in the murine forebrain. Neuron 26, 395–404 (2000).

    Article  CAS  Google Scholar 

  19. Ishibashi, M. et al. Persistent expression of helix-loop-helix factor HES-1 prevents mammalian neural differentiation in the central nervous system. EMBO J. 13, 1799–1805 (1994).

    Article  CAS  Google Scholar 

  20. Mizutani, K. & Saito, T. Progenitors resume generating neurons after temporary inhibition of neurogenesis by Notch activation in the mammalian cerebral cortex. Development 132, 1295–1304 (2005).

    Article  CAS  Google Scholar 

  21. Bajocchi, G., Feldman, S.H., Crystal, R.G. & Mastrangeli, A. Direct in vivo gene transfer to ependymal cells in the central nervous system using adenoviral vectors. Nat. Genet. 3, 229–234 (1993).

    Article  CAS  Google Scholar 

  22. Davidson, B.L. & Bohn, M.C. Recombinant adenovirus: a gene transfer vector for study and treatment of CNS disease. Exp. Neurol. 144, 125–130 (1997).

    Article  CAS  Google Scholar 

  23. Akli, S. et al. Transfer of a foreign gene into the brain using adenovirus vectors. Nat. Genet. 3, 224–228 (1993).

    Article  CAS  Google Scholar 

  24. Ostrowski, L.E., Hutchins, J.R., Zakel, K. & O'Neal, W.K. Targeting expression of a transgene to the airway surface epithelium using a ciliated cell–specific promoter. Mol. Ther. 8, 637–645 (2003).

    Article  CAS  Google Scholar 

  25. Lim, L., Zhou, H. & Costa, R.H. The winged helix transcription factor HFH-4 is expressed during choroid plexus epithelial development in the mouse embryo. Proc. Natl. Acad. Sci. USA 94, 3094–3099 (1997).

    Article  CAS  Google Scholar 

  26. Blatt, E.N., Yan, X.H., Wuerffel, M.K., Hamilos, D.L. & Brody, S.L. Forkhead transcription factor HFH-4 expression is temporally related to ciliogenesis. Am. J. Respir. Cell Mol. Biol. 21, 168–176 (1999).

    Article  CAS  Google Scholar 

  27. Soriano, P. Generalized lacZ expression with the Rosa26-cre reporter strain. Nat. Genet. 21, 70–71 (1999).

    Article  CAS  Google Scholar 

  28. Lindvall, O. & Kokaia, Z. Neurogenesis following stroke affecting the adult brain. in Adult Neurogenesis (eds. Gage, F.H., Kempermann, G. & Song, H.) (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 2008).

    Google Scholar 

  29. Barnabe-Heider, F. et al. Genetic manipulation of adult mouse neurogenic niches by in vivo electroporation. Nat. Methods 5, 189–196 (2008).

    Article  CAS  Google Scholar 

  30. Sang, L., Coller, H.A. & Roberts, J.M. Control of the reversibility of cellular quiescence by the transcriptional repressor HES1. Science 321, 1095–1100 (2008).

    Article  CAS  Google Scholar 

  31. Wei, Y., Yu, L., Bowen, J., Gorovsky, M.A. & Allis, C.D. Phosphorylation of histone H3 is required for proper chromosome condensation and segregation. Cell 97, 99–109 (1999).

    Article  CAS  Google Scholar 

  32. Lledo, P.M., Alonso, M. & Grubb, M.S. Adult neurogenesis and functional plasticity in neuronal circuits. Nat. Rev. Neurosci. 7, 179–193 (2006).

    Article  CAS  Google Scholar 

  33. Winner, B., Cooper-Kuhn, C.M., Aigner, R., Winkler, J. & Kuhn, H.G. Long-term survival and cell death of newly generated neurons in the adult rat olfactory bulb. Eur. J. Neurosci. 16, 1681–1689 (2002).

    Article  Google Scholar 

  34. Androutsellis-Theotokis, A. et al. Notch signaling regulates stem cell numbers in vitro and in vivo. Nature 442, 823–826 (2006).

    Article  CAS  Google Scholar 

  35. Dor, Y. & Melton, D.A. Facultative endocrine progenitor cells in the adult pancreas. Cell 132, 183–184 (2008).

    Article  CAS  Google Scholar 

  36. Dor, Y., Brown, J., Martinez, O.I. & Melton, D.A. Adult pancreatic beta-cells are formed by self-duplication rather than stem-cell differentiation. Nature 429, 41–46 (2004).

    Article  CAS  Google Scholar 

  37. Simon, A. & Frisen, J. From stem cell to progenitor and back again. Cell 128, 825–826 (2007).

    Article  CAS  Google Scholar 

  38. Nakagawa, T., Nabeshima, Y. & Yoshida, S. Functional identification of the actual and potential stem cell compartments in mouse spermatogenesis. Dev. Cell 12, 195–206 (2007).

    Article  CAS  Google Scholar 

  39. Levy, V., Lindon, C., Harfe, B.D. & Morgan, B.A. Distinct stem cell populations regenerate the follicle and interfollicular epidermis. Dev. Cell 9, 855–861 (2005).

    Article  CAS  Google Scholar 

  40. Ito, M. et al. Stem cells in the hair follicle bulge contribute to wound repair, but not to homeostasis of the epidermis. Nat. Med. 11, 1351–1354 (2005).

    Article  CAS  Google Scholar 

  41. Xu, X. et al. Beta cells can be generated from endogenous progenitors in injured adult mouse pancreas. Cell 132, 197–207 (2008).

    Article  CAS  Google Scholar 

  42. Tanigaki, K. et al. Notch–RBP-J signaling is involved in cell fate determination of marginal zone B cells. Nat. Immunol. 3, 443–450 (2002).

    Article  CAS  Google Scholar 

  43. Consiglio, A. et al. Robust in vivo gene transfer into adult mammalian neural stem cells by lentiviral vectors. Proc. Natl. Acad. Sci. USA 101, 14835–14840 (2004).

    Article  CAS  Google Scholar 

  44. Hara, H., Huang, P.L., Panahian, N., Fishman, M.C. & Moskowitz, M.A. Reduced brain edema and infarction volume in mice lacking the neuronal isoform of nitric oxide synthase after transient MCA occlusion. J. Cereb. Blood Flow Metab. 16, 605–611 (1996).

    Article  CAS  Google Scholar 

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Acknowledgements

We are grateful to U. Lendahl, O. Hermanson, A. Simon and members of the Frisén laboratory for valuable discussions; L. Ostrowski for the Foxj1 promoter construct; T. Li for antibodies to Crocc; U. Lendahl for N1IC and 12× Rbpj-dsgfp cDNA; O. Bergmann for virus injections; and M-L. Spångberg and K. Hamrin for technical assistance. This study was supported by grants from the Swedish Research Council, the Swedish Cancer Society, the Foundation for Strategic Research, the Juvenile Diabetes Research Foundation, the Karolinska Institute, Tobias Stiftelsen and the European Commission Framework VI Programme, EuroStemCell and StemStroke. M.C. and E.E. are supported by postdoctoral fellowships from the Swedish Brain Foundation (Hjärnfonden), C.G. by a postdoctoral fellowship from the Wenner-Gren Foundation and F.B.-H. by postdoctoral fellowships from the Canadian Institutes of Health Research and the Christopher and Dana Reeve Foundation.

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  1. Marie Carlén & Konstantinos Meletis

    Present address: Present address: Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.,

Authors and Affiliations

  1. Department of Cell and Molecular Biology, Medical Nobel Institute, Karolinska Institute, Stockholm, SE-171 77, Sweden

    Marie Carlén, Konstantinos Meletis, Christian Göritz, Fanie Barnabé-Heider, Maggie S Y Yeung, Robert M Cassidy & Jonas Frisén

  2. Wallenberg Neuroscience Center, University Hospital, Lund, SE-221 84, Sweden

    Vladimer Darsalia, Zaal Kokaia & Olle Lindvall

  3. Lund Strategic Research Center for Stem Cell Biology and Cell Therapy, University Hospital, Lund, SE-221 84, Sweden

    Vladimer Darsalia, Zaal Kokaia & Olle Lindvall

  4. Department of Neuroscience, Karolinska Institute, Stockholm, SE-171 77, Sweden

    Emma Evergren & Oleg Shupliakov

  5. Department of Medical Chemistry, Graduate School of Medicine, Kyoto University, Konoecho, Yoshida, Sakyo-ku, Kyoto, 606-8501, Japan

    Kenji Tanigaki & Tasuku Honjo

  6. Telethon Institute for Gene Therapy, San Raffaele Institute, via Olgettina 58, Milan, 20132, Italy

    Mario Amendola & Luigi Naldini

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Contributions

M.C. and K.M. designed the study, carried out viral injections, analyzed and documented the Rbpj ablation and stroke experiments and wrote the paper. C.G. carried out viral injections and analyzed and documented the stroke experiments. V.D. conducted the stroke experiments. E.E. performed ultrastructural analyses. K.T. helped with the initial Rbpj ablation experiments. M.A. produced lentivirus. F.B.-H. carried out electroporation. M.S.Y.Y. contributed to the documentation of neuronal maturation in the olfactory bulb. L.N. supervised the lentivirus production. T.H. provided conditional Rbjp mice. Z.K. designed and analyzed the stroke experiments. O.S. designed and performed the ultrastructural analyses. R.M.C. initiated the study, carried out viral injections, analyzed and documented the Rbpj ablation experiments and the expression of components of the Notch pathway. O.L. designed and supervised the stroke experiments and wrote the paper. J.F. designed and supervised the study and wrote the paper.

Corresponding author

Correspondence to Jonas Frisén.

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Carlén, M., Meletis, K., Göritz, C. et al. Forebrain ependymal cells are Notch-dependent and generate neuroblasts and astrocytes after stroke. Nat Neurosci 12, 259–267 (2009). https://doi.org/10.1038/nn.2268

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