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SOX9 induces and maintains neural stem cells

Abstract

Neural stem cells (NSCs) are uncommitted cells of the CNS defined by their multipotentiality and ability to self renew. We found these cells to not be present in substantial numbers in the CNS until after embryonic day (E) 10.5 in mouse and E5 in chick. This coincides with the induction of SOX9 in neural cells. Gain- and loss-of-function studies indicated that SOX9 was essential for multipotent NSC formation. Moreover, Sonic Hedgehog was able to stimulate precocious generation of NSCs by inducing Sox9 expression. SOX9 was also necessary for the maintenance of multipotent NSCs, as shown by in vivo fate mapping experiments in the adult subependymal zone and olfactory bulbs. In addition, loss of SOX9 led ependymal cells to adopt a neuroblast identity. These data identify a functional link between extrinsic and intrinsic mechanisms of NSCs specification and maintenance, and establish a central role for SOX9 in the process.

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Figure 1: Acquisition of neurosphere forming ability in mouse and chick CNS and the effect of SHH signaling.
Figure 2: The expression of SOX9 coincides with the formation of NSCs.
Figure 3: Ectopic Sox9 can induce precocious neurosphere formation from embryonic CNS and increase neurosphere formation from the adult SEZ.
Figure 4: Sox9 is necessary for neurosphere formation.
Figure 5: Sox9 is necessary for neurosphere formation at E18.5, and required for multipotentiality in vitro and in vivo.
Figure 6: SOX9 and SOX10 mediate the effects of SHH signaling.
Figure 7: Sox9 is necessary for NSC function in vivo in the adult.
Figure 8: Sox9 is necessary for NSC function in vivo in the adult olfactory bulb.

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Acknowledgements

We thank A. Schedl (INSERM U636, Nice) for the conditional Sox9 mutant mice, A. Nagy (Samuel Lunenfeld Research Institute) for the pCall2 vector, W.C.W. Chan for blastocyst injections and production of Z/Sox9 mice and T. Caspary (Emory University) for the Arl13b antibody. The CMV:cre construct was a gift from S. O'Gorman (Salk Institute). Thank you to S. Guioli, F. Guillemot and L. Reynard for critical reading of the manuscript, to C. Andoniadou for advice and training in the ways of neurosphere cultures, to W. Han Yau in the photographics department at NIMR for help with illustrations, to T. Matabanadzo and other biological services staff at NIMR for help with the mouse colonies and other members of our laboratories for discussion and encouragement. This work was supported by the UK Medical Research Council (U117512772), a US National Institutes of Health (National Institute of Biomedical Imaging and Bioengineering) Quantum Grant (R.L.-B. and C.E.S.), and grants from the Hong Kong Research Grants Council and the Hong Kong University Grants Council Area of Excellence Scheme (S.L.W. and K.S.E.C.).

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C.E.S., J.B. and R.L.-B. initiated the project. C.E.S. performed the in ovo electroporations, neurosphere culturing, immunohistochemistry, RT-PCR and quantification and data analysis of all the in vivo and in vitro experiments, except for the acquisition and analysis of the microarray data (C.C.). S.L.W., B.G. and K.S.E.C. generated the Z/Sox9 mice. M.C. supplied many of the constructs used in this study. A.S. performed the adenovirus injections and S.B. perfused the adult mice. M.-V.G.G. carried out the BrdU injections. C.E.S., R.L.-B. and J.B. were involved in the study design and wrote the manuscript.

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Correspondence to Robin Lovell-Badge or James Briscoe.

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Scott, C., Wynn, S., Sesay, A. et al. SOX9 induces and maintains neural stem cells. Nat Neurosci 13, 1181–1189 (2010). https://doi.org/10.1038/nn.2646

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