jhdm1d基因在小鼠胚胎中的表达及功能分析-expression and functional analysis of jhd m1 d gene in mouse embryos.docxVIP

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jhdm1d基因在小鼠胚胎中的表达及功能分析-expression and functional analysis of jhd m1 d gene in mouse embryos.docx

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jhdm1d基因在小鼠胚胎中的表达及功能分析-expression and functional analysis of jhd m1 d gene in mouse embryos

AbstractSince 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. Jhdm1d, encoding a histone demethylase, belongs to a member of Jumonji family containing the JmjC domain. However the spatiotemporal expression patterns and function of Jhdm1d in mouse embryonic development have not been systematically studied.In this study, we analyzed the expression patterns of Jhdm1d in mouse differentdevelopmental stages in detail by quantitative real-time PCR (qRT-PCR), western blot,and immunohistochemistry(IHC). The epigenetic regulation mechanism (DNA methylation) of Jhdm1d was analyzed by using bisulfite sequencing and its functions were analyzed in mouse C2C12 cells. 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 Jhdm1d was gradually increased from oocyte to 8-cells and then gradually decreased from 8-cells to blastocyst. The Jhdm1d expression was the highest in 8-cells and the lowest in blastocyst. In the mid-later gestation period, the expression of Jhdm1d was significantly different in various tissues. At E12.5, Jhdm1d was expressed abundantly in brain and liver, weakly in heart and kidney, accompanied by the lowest level in tongue and lung. At E15.5 Jhdm1d 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 Jhdm1d in various tissues were almost very low. The immunohistochemistry results at E12.5 and E15.5 were consistent with qRT-PCR. At the same time, the qRT-PCR showed that Jhdm1d were strongly increased during C2C12 differentiation. The expression was the lowest at D0, and reached the highest at D7. The results of RNAi in C2C12 suggested that Jhdm1d expression was