Mammalian Gcm genes induce Hes5 expression by active DNA demethylation and induce neural stem cells

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Signaling mediated by Notch receptors is crucial for the development of many organs and the maintenance of various stem cell populations. The activation of Notch signaling is first detectable by the expression of an effector gene, Hes5, in the neuroepithelium of mouse embryos at embryonic day (E) 8.0–8.5, and this activation is indispensable for the generation of neural stem cells. However, the molecular mechanism by which Hes5 expression is initiated in stem-producing cells remains unknown. We found that mammalian Gcm1 and Gcm2 (glial cells missing 1 and 2) are involved in the epigenetic regulation of Hes5 transcription by DNA demethylation independently of DNA replication. Loss of both Gcm genes and subsequent lack of Hes5 upregulation in the neuroepithelium of E7.5–8.5 Gcm1−/−; Gcm2−/− mice resulted in the impaired induction of neural stem cells. Our data suggest that Hes5 expression is serially activated first by Gcms and later by the canonical Notch pathway.

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Figure 1: DNA methylation in the Hes5 promoter.
Figure 2: Gcms are responsible for the demethylation of the Hes5 promoter.
Figure 3: Gcms are indispensable for Hes5 induction and neural stem cell generation.
Figure 4: Abnormal neural development in Gcm2 null mutants.
Figure 5: Gcms induce Hes5 expression in embryonic brains.
Figure 6: Gcms demethylate mitotically inactive DNA and methylated plasmids.
Figure 7: Active demethylation by Gcms in vivo.


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We thank R. Kageyama, J.S. Nye, T. Miyazaki and T. Honjo for plasmids, T.-e.U and Y. Imai for MBD4 mutant mice, and R. Kageyama, D. van der Kooy and K. Nakashima for comments. This work was supported by a Grant-in-Aid for Exploratory Research (19650096) from the Ministry of Education, Culture, Sports, Science and Technology of Japan (K.I.S.H.).

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S.H. designed and carried out the experiments, analyzed the data and wrote the paper. Y.I., A.K., S.J., K.F.T. and T.H. generated Gcm mutant mice and analyzed the phenotypes. T.K. and S.K. carried out the experiments related to MBD4 knockout mice. Y.H. and K.I. supervised the project.

Correspondence to Seiji Hitoshi.

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Hitoshi, S., Ishino, Y., Kumar, A. et al. Mammalian Gcm genes induce Hes5 expression by active DNA demethylation and induce neural stem cells. Nat Neurosci 14, 957–964 (2011) doi:10.1038/nn.2875

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