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miR-124 regulates adult neurogenesis in the subventricular zone stem cell niche

Nature Neuroscience volume 12, pages 399–408 (2009)Cite this article

Abstract

The subventricular zone (SVZ) is the largest neurogenic niche in the adult mammalian brain. We found that the brain-enriched microRNA miR-124 is an important regulator of the temporal progression of adult neurogenesis in mice. Knockdown of endogenous miR-124 maintained purified SVZ stem cells as dividing precursors, whereas ectopic expression led to precocious and increased neuron formation. Furthermore, blocking miR-124 function during regeneration led to hyperplasias, followed by a delayed burst of neurogenesis. We identified the SRY-box transcription factor Sox9 as being a physiological target of miR-124 at the transition from the transit amplifying cell to the neuroblast stage. Sox9 overexpression abolished neuronal differentiation, whereas Sox9 knockdown led to increased neuron formation. Thus miR-124–mediated repression of Sox9 is important for progression along the SVZ stem cell lineage to neurons.

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Figure 1: miR-124 expression in the adult SVZ niche.
Figure 2: Knockdown of miR-124 in vitro.
Figure 3: Ectopic expression of miR-124 in vitro.
Figure 4: miR-124 overexpression and knockdown in vivo.
Figure 5: miR-124 knockdown delays SVZ regeneration.
Figure 6: Sox9 is a miR-124 target.
Figure 7: Effect of miR-124 knockdown on Sox9 protein levels.
Figure 8: Sox9 knockdown induces neuronal differentiation.

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Acknowledgements

We thank K. Gordon for assistance with FACS at the Herbert Irving Comprehensive Cancer Center at Columbia University. We thank members of the Doetsch and Wichterle laboratories for critical reading of the manuscript. We are grateful to C. Troy for advice on penetratin 1–mediated delivery, N. Heintz for antibody to BLBP and R. Tsien for pCMV-Cherry plasmid. The nestin, M2 and M6 antibodies developed by S. Hockfield and C. Lagenaur, respectively, were obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the National Institute of Child Health and Human Development and maintained by the University of Iowa. This work was supported by a US National Institutes of Health National Institute of Neurological Disorders and Stroke grant to F.D. E.P. was supported by a US National Institutes of Health training grant and a grant from the Spanish Ministerio de Educacion y Ciencia. M.T. was supported by a Medical Scientist Training Program fellowship from the US National Institutes of Health, F.D. is a Packard Foundation Fellow and an Irma T. Hirschl Fellow, and this work was partially supported by the Jerry and Emily Spiegel Laboratory for Cell Replacement Therapies and the Anne and Bernard Spitzer Fund for Cell Replacement Therapy. We dedicate this article to the memory of Emily Spiegel.

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Authors and Affiliations

  1. Departments of Pathology and Cell Biology Columbia University, College of Physicians and Surgeons, New York, New York, USA

    Li-Chun Cheng, Erika Pastrana & Fiona Doetsch

  2. Department of Neuroscience, Columbia University, College of Physicians and Surgeons, New York, New York, USA

    Masoud Tavazoie & Fiona Doetsch

  3. Department of Neurology, Columbia University, College of Physicians and Surgeons, New York, New York, USA

    Fiona Doetsch

  4. Center for Motor Neuron Biology and Disease, Columbia University, College of Physicians and Surgeons, New York, New York, USA

    Fiona Doetsch

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  1. Li-Chun Cheng
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Contributions

L.-C.C. and F.D. initiated the project. L.-C.C. generated the pSUPERretro (pSR)-GFP, pSR-124mt and pSR-shRNA constructs. L.-C.C. and M.T. generated the pSR-124 construct. M.T. and E.P. generated the Sox9 overexpression construct. L.-C.C. performed the in situ hybridization, qRT-PCR, neurosphere assays, neuron survival assays, Sox9 expression assay, retroviral production, penetratin conjugation, in vivo delivery and Ara-C experiments. L.-C.C. and E.P. carried out the FACS sorting, cocultures, adherent cultures and in vivo injections. E.P. performed the Gene Ontology studies. M.T. carried out the luciferase assays and Sox9 immunohistochemistry. F.D. was involved in the study design, data collection, quantification and data analysis. The manuscript was written by F.D., L.-C.C. and E.P. All authors performed data quantification, discussed the results and commented on the manuscript.

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Correspondence to Fiona Doetsch.

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Supplementary Figures 1–8, Supplementary Tables 1 and 2, and Supplementary Methods (PDF 2797 kb)

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Cheng, LC., Pastrana, E., Tavazoie, M. et al. miR-124 regulates adult neurogenesis in the subventricular zone stem cell niche. Nat Neurosci 12, 399–408 (2009). https://doi.org/10.1038/nn.2294

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