The histone methyltransferase ESET is required for the survival of spermatogonial stem/progenitor cells in mice.
Self-renewal and differentiation of spermatogonial stem cells (SSCs) are the foundation of spermatogenesis throughout a male's life. SSC transplantation will be a valuable solution for young male patients to preserve their fertility. As SSCs in the collected testis tissue from the patients are very limited, it is necessary to expansion the SSCs in vitro. Previous studies suggested that histone methyltransferase ERG-associated protein with SET domain (ESET) represses gene expression and is essential for the maintenance of the pool of embryonic stem cells and neurons. The objective of this study was to determine the role of ESET in SSCs using in vitro cell culture and germ cell transplantation. Cell transplantation assay showed that knockdown of ESET reduced the number of seminiferous tubules with spermatogenesis when compared with that of the control. Knockdown of ESET also upregulated the expression of apoptosis-associated genes (such as P53, Caspase9, Apaf1), whereas inhibited the expression of apoptosis-suppressing genes (such as Bcl2l1, X-linked inhibitor of apoptosis protein). In addition, suppression of ESET led to increase in expression of Caspase9 and activation of Caspase3 (P17) as well as cleavage of poly (ADP-ribose) polymerase. Among the five ESET-targeting genes (Cox4i2, spermatogenesis and oogenesis Specific Basic Helix-Loop-Helix 2, Nobox, Foxn1 and Dazl) examined by ChIP assay, Cox4i2 was found to regulate SSC apoptosis by the rescue experiment. BSP analyses further showed that DNA methylation in the promoter loci of Cox4i2 was influenced by ESET, indicating that ESET also regulated gene expression through DNA methylation in addition to histone methylation. In conclusion, we found that ESET regulated SSC apoptosis by suppressing of Cox4i2 expression through histone H3 lysine 9 tri-methylation and DNA methylation. The results obtained will provide unique insights that would broaden the research on SSC biology and contribute to the treatment of male infertility.
College of Animal Science and Technology, Northwest A&F University, Shaanxi, China.
J
J
An
College of Animal Science and Technology, Northwest A&F University, Shaanxi, China.
X
X
Zhang
College of Animal Science and Technology, Northwest A&F University, Shaanxi, China.
J
J
Qin
College of Animal Science and Technology, Northwest A&F University, Shaanxi, China.
Y
Y
Wan
College of Animal Science and Technology, Northwest A&F University, Shaanxi, China.
Y
Y
Hu
College of Animal Science and Technology, Northwest A&F University, Shaanxi, China.
T
T
Liu
College of Animal Science and Technology, Northwest A&F University, Shaanxi, China.
J
J
Li
College of Animal Science and Technology, Northwest A&F University, Shaanxi, China.
W
W
Dong
College of Veterinary Medicine, Northwest A&F University, Shaanxi, China.
E
E
Du
College of Animal Science and Technology, Northwest A&F University, Shaanxi, China.
C
C
Pan
College of Animal Science and Technology, Northwest A&F University, Shaanxi, China.
W
W
Zeng
D000818:Animals; D017209:Apoptosis; D002470:Cell Survival; D002478:Cells, Cultured; D019175:DNA Methylation; D003576:Electron Transport Complex IV / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D011495:Histone-Lysine N-Methyltransferase / Q000378:metabolism*; D006657:Histones / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D008297:Male; D008745:Methylation; D008810:Mice, Inbred C57BL; D008954:Models, Biological; D010641:Phenotype; D034622:RNA Interference; D013093:Spermatogonia / Q000166:cytology*; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000201:enzymology*; D013737:Testis / Q000201:enzymology; D014161:Transduction, Genetic
D006657:Histones; D003576:Electron Transport Complex IV; D011495:Histone-Lysine N-Methyltransferase; C516239:SETDB1 protein, mouse; D008239:Lysine