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Critical Role of Tet3 in Neural Progenitor Cell Maintenance and Terminal Differentiation

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Abstract

5-Hydroxymethylcytosine (5hmC), converted from 5-methylcytocine (5mC) by Tet family of dioxygenases (Tet1, Tet2, and Tet3), is enriched in the embryonic stem cells (ESCs) and in the brain. However, the role of 5hmC and Tet family in the process of ESC differentiation especially neuronal differentiation remains elusive. Here, we showed the evidence that Tet3 is critical in neural progenitor cell (NPC) maintenance and terminal differentiation of neurons. We found that Tet3 expression is basically undetectable in ESCs, but its level increases rapidly during neuronal differentiation. Tet3 knockout ESCs appear normal in self-renewal and maintenance but impaired in neuronal differentiation. NPCs could be induced efficiently from Tet3 knockout ESCs, as the expression of NPC marker Pax6 and nestin is comparable with NPCs from wild-type ESCs, but undergo apoptosis rapidly, and the terminal differentiation of neurons is greatly reduced. Our results indicate that Tet3 is important for NPC maintenance and terminal differentiation of neurons.

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Acknowledgements

We thank Guoliang Xu for helpful discussions. This work was funded by the National Natural Sciences Foundation of China (No. 81171201) and the National Basic Research Program of China (No. 2011CB510003).

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The authors declare no competing financial interests.

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Authors and Affiliations

  1. Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200025, China

    Ting Li, Dehua Yang, Jia Li, Yu Tang, Juan Yang & Weidong Le

  2. Center for Translational Research of Neurology Disease, First Affiliated Hospital, Dalian Medical University School of Medicine, Dalian, 116011, China

    Weidong Le

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  1. Ting Li
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Supplementary Fig. S1

The mRNA levels of Tet family genes and 5hmC levels in differentiated Tet3 KO cells. (a) Quantitative RT-PCR for Tet1, Tet2, and Tet3 mRNA in wild-type (WT) and knockout cells differentiated for 8 days. Data are means ± s.e.m. for three independent experiments. (b) Dot blots of 5hmC. Genomic DNA was extracted from WT or knockout cells differentiated for 8 days. 2-fold serial dilutions of genomic DNA was treated and loaded on the membrane for further detection. (GIF 35 kb)

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Li, T., Yang, D., Li, J. et al. Critical Role of Tet3 in Neural Progenitor Cell Maintenance and Terminal Differentiation. Mol Neurobiol 51, 142–154 (2015). https://doi.org/10.1007/s12035-014-8734-5

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