Open AccessModification of Tet1 and histone methylation dynamics in dairy goat male germline stem cells
Author(s) -
Zheng Liming,
Zhai Yuanxin,
Li Na,
Wu Chongyang,
Zhu Haijing,
Wei Zhuying,
Bai Chunling,
Li Guangpeng,
Hua Jinlian
Publication year - 2016
Publication title -
cell proliferation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.647
H-Index - 74
eISSN - 1365-2184
pISSN - 0960-7722
DOI - 10.1111/cpr.12245
Subject(s) - germline , histone , biology , methylation , dna methylation , stem cell , dynamics (music) , epigenetics , microbiology and biotechnology , genetics , dna , gene , gene expression , physics , acoustics
Objectives Tet (ten‐eleven translocation) protein 1 is a key enzyme for DNA demethylation, which modulates DNA methylation and gene transcription. DNA methylation and histone methylation are critical elements in self‐renewal of male germline stem cells ( mGSC s) and spermatogenesis. mGSC s are the only type of adult stem cells able to achieve intergenerational transfer of genetic information, which is accomplished through differentiated sperm cells. However, numerous epigenetic obstacles including incomplete DNA methylation and histone methylation dynamics make establishment of stable livestock mGSC cell lines difficult. The present study was conducted to detect effects of DNA methylation and histone methylation dynamics in dairy goat mGSC s self‐renewal and proliferation, through overexpression of Tet1. Materials and methods An immortalized dairy goat mGSC cell line bearing mouse Tet1 ( mT et1 ) gene was screened and characteristics of the cells were assayed by quantitative real‐time PCR ( qRT ‐ PCR ), immunofluorescence assay, western blotting, fluorescence activated cell sorting ( FACS ) and use of the cell counting kit ( CCK 8) assay. Results The screened immortalized dairy goat mGSC cell line bearing mT et1 , called mGSC ‐ mT et1 cells was treated with optimal doxycycline (Dox) concentration to maintain Tet1 gene expression. mGSC ‐ mT et1 cells proliferated at a significantly greater rate than wild‐type mGSC s, and mGSC s‐specific markers such as proliferating cell nuclear antigen ( PCNA ), cyclinD1 ( CCND 1), GDNF family receptor alpha 1 (Gfra1) and endogenic Tet1, Tet2 were upregulated. The cells exhibited not only reduction in level of histone methylation but also changes in nuclear location of that methylation marker. While H3K9me3 was uniformly distributed throughout the nucleus of mGSC ‐ mT et1 cells, it was present in only particular locations in mGSC s. H3K27me3 was distributed surrounding the edges of nuclei of mGSC ‐ mT et1 cells, while it was uniformly distributed throughout nuclei of mGSC s. Our results conclusively demonstrate that modification of mGSC s with mT et1 affected mGSC maintenance and seemed to promote establishment of stable goat mGSC cell lines. Conclusions Taken together, our data suggest that Tet1 had novel and dynamic roles for regulating maintenance of pluripotency and proliferation of mGSC s by forming complexes with PCNA and histone methylation dynamics. This may provide new solutions for mGSC s stability and livestock mGSC cell line establishment.