Regulation of pluripotency in male germline stem cells by Dmrt1
- Seiji Takashima1,
- Michiko Hirose2,
- Narumi Ogonuki2,
- Miki Ebisuya3,
- Kimiko Inoue2,
- Mito Kanatsu-Shinohara1,
- Takashi Tanaka1,
- Eisuke Nishida4,
- Atsuo Ogura2 and
- Takashi Shinohara1,5,6
- 1Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan;
- 2RIKEN Bioresource Center, Tsukuba 305-0074, Japan;
- 3Career-Path Promotion Unit for Young Life Scientists, Kyoto University, Kyoto 606-8501, Japan;
- 4Department of Cell and Developmental Biology, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan;
- 5Japan Science and Technology Agency, CREST, Kyoto 606-8501, Japan
Abstract
Spermatogonial stem cells (SSCs) present the potential to acquire pluripotency under specific culture conditions. However, the frequency of pluripotent cell derivation is low, and the mechanism of SSC reprogramming remains unknown. In this study, we report that induction of global DNA hypomethylation in germline stem (GS) cells (cultured SSCs) induces pluripotent cell derivation. When DNA demethylation was triggered by Dnmt1 depletion, GS cells underwent apoptosis. However, GS cells were converted into embryonic stem (ES)-like cells by double knockdown of Dnmt1 and p53. This treatment down-regulated Dmrt1, a gene involved in sexual differentiation, meiosis, and pluripotency. Dmrt1 depletion caused apoptosis of GS cells, but a combination of Dmrt1 and p53 depletion also induced pluripotency. Functional screening of putative Dmrt1 target genes revealed that Dmrt1 depletion up-regulates Sox2. Sox2 transfection up-regulated Oct4 and produced pluripotent cells. This conversion was enhanced by Oct1 depletion, suggesting that the balance of Oct proteins maintains SSC identity. These results suggest that spontaneous SSC reprogramming is caused by unstable DNA methylation and that a Dmrt1–Sox2 cascade is critical for regulating pluripotency in SSCs.
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Footnotes
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↵6 Corresponding author
E-mail tshinoha{at}virus.kyoto-u.ac.jp
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Supplemental material is available for this article.
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Article published online ahead of print. Article and publication date are online at http://www.genesdev.org/cgi/doi/10.1101/gad.220194.113.
- Received April 22, 2013.
- Accepted August 23, 2013.
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