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Hippocampal development and neural stem cell maintenance require Sox2-dependent regulation of Shh

Nature Neuroscience volume 12, pages 1248–1256 (2009)Cite this article

Abstract

Neural stem cells (NSCs) are controlled by diffusible factors. The transcription factor Sox2 is expressed by NSCs and Sox2 mutations in humans cause defects in the brain and, in particular, in the hippocampus. We deleted Sox2 in the mouse embryonic brain. At birth, the mice showed minor brain defects; shortly afterwards, however, NSCs and neurogenesis were completely lost in the hippocampus, leading to dentate gyrus hypoplasia. Deletion of Sox2 in adult mice also caused hippocampal neurogenesis loss. The hippocampal developmental defect resembles that caused by late sonic hedgehog (Shh) loss. In mutant mice, Shh and Wnt3a were absent from the hippocampal primordium. A SHH pharmacological agonist partially rescued the hippocampal defect. Chromatin immunoprecipitation identified Shh as a Sox2 target. Sox2-deleted NSCs did not express Shh in vitro and were rapidly lost. Their replication was partially rescued by the addition of SHH and was almost fully rescued by conditioned medium from normal cells. Thus, NSCs control their status, at least partly, through Sox2-dependent autocrine mechanisms.

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Figure 1: Sox2 conditional null allele, SOX2 protein ablation by nestin-Cre in mutant embryonic brain and in vivo morphological defects of nestin-cre Sox2-deleted mutants (Sox2null).
Figure 2: Cellular defects of nestin-cre Sox2-deleted mutant brain.
Figure 3: Sox2-deleted mutant brains have defective Shh and Wnt3a mRNA expression.
Figure 4: SHH colocalizes with SOX2 in neural cells in postnatal neurogenic regions and its expression is lost in Sox2null mutants.
Figure 5: Stimulation of the Shh signaling pathway rescues hippocampal NSC and neurogenesis in Sox2 mutants.
Figure 6: Sox2 deletion in adult brain leads to rapid loss of radial glia cells and of cell proliferation in the hippocampus dentate gyrus.
Figure 7: Impaired maintenance of Sox2null NSCs in culture and rescue by extracellular factors.
Figure 8: Shh is a direct target of SOX2.

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Acknowledgements

We thank G. Dehò and S. Zangrossi for the pcnB∷Tn10 recA∷cat C-5507 E. coli C strain, S. Dymecki for the pLM-FLRT-3 vector and the FLPeR mouse strain, R. Klein through Jackson Laboratories for the nestin-cre mouse strain, P. Chambon for the creERT2 gene, Curis for the 1.2 SHH agonist18, S. Lugert for help with the Notch pathway expression studies, J. Cozzi and GenOway for help with ES cell targeting, C. Tiveron and Layline Genomics for blastocyst injections, C. Tiveron and L. Tatangelo for Sox2-creERT2 pronuclear injections, R. Caccia for help with in situ hybridization, E. Bagnaresi and A. Marinelli for help with constructs, A. Ronchi for advice on EMSAs and ChIP and D. Santoni for animal care. The monoclonal antibody to SHH developed by T.M. Jessell was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the US National Institute of Child Health and Human Development and maintained by the University of Iowa. This work was supported by Telethon (GGP05122), the European Economic Community (STEMBRIDGE), Fondazione Cariplo, Associazione Italiana Ricerca sul Cancro, Fondazione Banca del Monte di Lombardia, the Ministero Istruzione Università e Ricerca (Cofin) and FAR 2004-7 grants to S.K.N., and by the Max Planck Gesellschaft through V. Taylor.

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Author notes

  1. Menella Valotta

    Present address: Present address: Axxam, Milano, Italy.,

  2. Menella Valotta and Anna L M Ferri: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milano, Italy

    Rebecca Favaro, Menella Valotta, Anna L M Ferri, Elisa Latorre, Jessica Mariani, Cesare Lancini, Valentina Tosetti, Sergio Ottolenghi & Silvia K Nicolis

  2. Department of Molecular Embryology, Max Planck Institute for Immunobiology, Freiburg im Breisgau, Germany

    Claudio Giachino & Verdon Taylor

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Contributions

R.F. and M.V. generated the Sox2 mutations in ES cells, and carried out animal breeding, cell cultures and RT-PCR. A.L.M.F. performed in situ hybridization and immunohistochemistry. E.L. carried out ChIP and EMSAs. J.M. performed the lentiviral rescue experiment. C.G. carried out the Notch signaling studies and initial work with in vitro cultures and for in vitro SOX2 deletion. C.L. participated in immunochemical studies. V. Tosetti participated in the in vitro culture work and immunocytochemistry. S.O. discussed the experiments and wrote the paper. V. Taylor participated in the in vivo rescue experiments, discussed data and supervised initial in vitro studies. S.K.N. devised the experiments, supervised the work and wrote the paper.

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Correspondence to Silvia K Nicolis.

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Favaro, R., Valotta, M., Ferri, A. et al. Hippocampal development and neural stem cell maintenance require Sox2-dependent regulation of Shh. Nat Neurosci 12, 1248–1256 (2009). https://doi.org/10.1038/nn.2397

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