title: Regulation of pluripotency in male germline stem cells by Dmrt1.
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.
authors.affiliation: Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan;
authors.forename: Seiji
authors.initials: S
authors.lastname: Takashima
authors.forename: Michiko
authors.initials: M
authors.lastname: Hirose
authors.forename: Narumi
authors.initials: N
authors.lastname: Ogonuki
authors.forename: Miki
authors.initials: M
authors.lastname: Ebisuya
authors.forename: Kimiko
authors.initials: K
authors.lastname: Inoue
authors.forename: Mito
authors.initials: M
authors.lastname: Kanatsu-Shinohara
authors.forename: Takashi
authors.initials: T
authors.lastname: Tanaka
authors.forename: Eisuke
authors.initials: E
authors.lastname: Nishida
authors.forename: Atsuo
authors.initials: A
authors.lastname: Ogura
authors.forename: Takashi
authors.initials: T
authors.lastname: Shinohara
mesh_terms: D000818:Animals; D002460:Cell Line; D065150:Cellular Reprogramming / Q000235:genetics; D019175:DNA Methylation; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D050811:Octamer Transcription Factor-1 / Q000235:genetics / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000378:metabolism / Q000502:physiology*; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D013093:Spermatogonia / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
chemical_list: C110547:DMRT1 protein; D050811:Octamer Transcription Factor-1; C494166:Pou2f1 protein, mouse; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse; D014157:Transcription Factors; D016159:Tumor Suppressor Protein p53
keywords: DNA methylation; pluripotency; reprogramming; spermatogonia; teratoma
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