Expression Patterns And Function Research Of Jhdm1dGENE During Mouse Embryo Development

Since the discovery of histone demethylases, more and more researchers have focused on them. Many reports have confirmed that histone demethylases involved in meiosis, cell differentiation and organ formation during the organisms development. J hdmld, encoding a histone demethylase, belongs to a member of Jumonji family containing the JmjC domain. However the spatiotemporal expression patterns and function of Jhdmld m mouse embryonic development have not been systematically studied.In this study, we analyzed the expression patterns of Jhdmld m mouse different developmental stages in detail by quantitative real-time PCR (qRT-PCR), western blot,and immunohistochemistry(IHC). The epigenetic regulation mechanism (DNA methylation) of Jhdmld was analyzed by using bisulfite sequencing and its functions were analyzed in mouse C2C12cells. This work might contribute to further study of regulation mechanism and functions of this gene.qRT-PCR results showed that on mouse preimplantation embryos, the expression of Jhdmld was gradually increased from oocyte to8-cells and then gradually decreased from8-cells to blastocyst. The Jhdmld expression was the highest in8-cells and the lowest in blastocyst. In the mid-later gestation period, the expression of Jhdmld was significantly different in various tissues. At E12.5, Jhdmld was expressed abundantly in brain and liver, weakly in heart and kidney, accompanied by the lowest level in tongue and lung. At E15.5Jhdmld was expressed abundantly in liver, weakly in brain, tongue, and heart, with the lowest level in lung and kidney. At E18.5, the expressions of Jhdmld in various tissues were almost very low. The immunohistochemistry results at E12.5and E15.5were consistent with qRT-PCR. At the same time, the qRT-PCR showed that Jhdmldwere strongly increased during C2C12differentiation. The expression was the lowest at DO, and reached the highest at D7. The results of RNAi in C2C12suggested that Jhdmld expression was knocked down about85%by shRNAl and no any knockdown effect by shRNA3. Additionally, knockdown of Jhdmld can result in the syndecan-1expression decrease. The results of bisulfite sequencing and the5-AZa treated in TM3cells demonstrated that Jhdmld expression was not regulated by DNA methylation. Together, these results suggest that Jhdmld may play an important role in the process involve in the whole embryonic developmental stages, and of C2C12differentiation.