Sequence analysis, expression patterns and transcriptional regulation of mouse Ifrg15 during preimplantation embryonic development

Wu, Fengrui; Ding, Biao; Qi, Bin; Shang, M.-B.; Yang, X.; Liu, Yong; Li, Wenyong

doi:10.1016/j.gene.2012.07.030

Cited by 5 publications

(7 citation statements)

References 30 publications

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“…All animals were maintained with a photoperiod of 14 h light and 10 h dark at 20–25 °C for at least 2 weeks before use. Zygotes, 2-cell, 4-cell and 8-cell embryos, morulae, and blastocysts were collected for this study as described previously [ 33 , 34 ]. All animal studies were approved by the Institutional Animal Care and Use Committee at Fuyang Teachers College in Anhui Province, China.…”

Section: Methodsmentioning

confidence: 99%

“…Fifty embryos were used for each time point, and three replicates were performed for each stage. Total RNA extraction, cDNA synthesis from all the samples, and real-time PCR were conducted according to the manufacturer’s instructions as described previously [ 33 ]. Gapdh was used as an internal control.…”

Section: Methodsmentioning

confidence: 99%

“…The threshold cycle (Ct) was defined as the fractional cycle number by the method of global minimum. The ratio change in Oct4 and Dnmt1 relative to the Gapdh control gene was determined using the 2 -△△Ct method [ 33 ]. Data are expressed as the means ± SE for the three replicates.…”

Section: Methodsmentioning

confidence: 99%

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Oct4 regulates DNA methyltransferase 1 transcription by direct binding of the regulatory element

et al. 2018

Cell Mol Biol Lett

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BackgroundThe transcription factor Oct4 plays a pivotal role in the pre-implantation development of the mouse embryo. DNA methyltransferase 1 (Dnmt1) maintains the changes in DNA methylation during mammalian early embryonic development. Little is known of the role of Oct4 in DNA methylation in mice. In this study, Kunming white mice were used as an animal model to reveal any correlation between DNA methylation and Oct4 during mammalian embryonic development.ResultsThe expressions of Dnmt1 and Oct4 were initially studied using real-time PCR. They exhibited different patterns during the pre-implantation stage. Moreover, by using a promoter assay and ChIP analysis, we found that the transcriptional activities of Dnmt1 in mouse NIH/3 T3 cells and CCE cells were regulated by Oct4 through direct binding to the − 554 to − 294 fragment of the upstream regulation element of Dnmt1. The downregulation of Dnmt1 expression and enzyme activity by mouse Oct4 were further confirmed by transfecting Oct4 siRNA into mouse CCE cells.ConclusionOur results indicate that Oct4 is involved in DNA methylation through the regulation of Dnmt1 transcription, especially during the early stages of mouse pre-implantation embryo development.Electronic supplementary materialThe online version of this article (10.1186/s11658-018-0104-2) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Oct4 regulates DNA methyltransferase 1 transcription by direct binding of the regulatory element

et al. 2018

Cell Mol Biol Lett

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“…It is well known that many development-related genes are upregulated in blastocysts compared to morulae (Tanaka and Ko, 2004). Demethylation of H3K27me3 during the blastocyst stage was synchronized with the modification of H3K4me3 (Wu et al, 2012a), indicating that many developmentrelated genes were activated in this stage to maintain the progress of embryonic development and implantation during the latter stage.…”

Section: Discussionmentioning

confidence: 98%

“…ab6002, Abcam, Cambridge, UK). Subsequent steps were also performed as described previously (Wu et al, 2012a). Samples without primary antibody were included as negative controls.…”

Section: Immunofluorescence Histochemistry For Detection Of H3k27me3mentioning

confidence: 99%

H3K27me3 may be associated with Oct4 and Sox2 in mouse preimplantation embryos

Zhang

Ding

et al. 2014

Genet. Mol. Res.

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ABSTRACT. As a core member of polycomb repressive complex 2, the transcription and enzyme activity of enhancer of zeste homolog 2 (Ezh2) is directly involved in the trimethylation of lysine 27 on histone H3. In this study, the fluorescence intensity of H3K27me3 in mouse in vivo morulae and blastocysts was compared by indirect immunofluorescence staining. We found that demethylation of H3K27me3 occurred during the blastocyst stage. Real-time polymerase chain reaction was performed to investigate Ezh2 expression in oocytes and in preimplantation embryos. Ezh2 expression peaked during the zygote stage and gradually decreased from the 2-cell stage, exhibiting an inverse pattern when compared with Oct4 and Sox2 mRNA in mouse preimplantation embryos. To understand the role of developmentrelated genes on the transcription of mouse Ezh2, a promoter assay was performed in NIH/3T3 cells. Ezh2 expression was markedly suppressed by Oct4 and Sox2 alone in a dose-dependent manner, while Ezh2 promoter activity in co-transfection with Nanog, Klf-4, and c-Myc groups showed no significant change as compared with the control. Our data suggest that the demethylation of H3K27me3 is caused by the degressive expression and activity of Ezh2 in blastocysts, leading to increased expression of developmentally important transcription factors. We also observed negative effects of Oct4 and Sox2 on the transcription of Ezh2 and identified Oct4 and Sox2 as novel negative regulators of Ezh2 at the post-translation level in a mouse preimplantation embryo.

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The interferon α-responsive gene, Ifrg15, plays vital roles during mouse early embryonic development

Yang

Wang

Zhao

et al. 2016

Cell. Mol. Life Sci.

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The interferon alpha-responsive gene (Ifrg15) mRNA is highly expressed in various stages during preimplantation mammalian embryo development. Unfortunately, few studies have investigated the effect of Ifrg15 in this process. In mammals, the fusion of male and female pronuclei generates a diploid zygote, and is an important step for subsequent cleavage and blastocyst formation. Here, by using RNA interference, rescue experiments, immunofluorescence staining and live cell observations, we found that preimplantation embryo development was arrested at the 1-cell stage after knocking down Ifrg15 expression. This induced DNA damage and prevented the cleavage of embryos. Furthermore, the effect of Ifrg15 deficiency in arresting preimplantation embryo development produced by specific short interfering RNA microinjection was concentration-dependent. Using transcriptome expression profiles, gene ontogeny functional annotation and enrichment analysis, we gained 197 enriched pathways based on 1445 differentially expressed genes (DEGs). Of these, 12 pathways and about one third of the DEGs were involved in DNA damage, DNA repair, cell cycle, and developmental processes. Thus, the IFRG15 protein might be an important molecule for maintaining genomic integrity and stability through upregulating or downregulating a cascade of genes to permit normal preimplantation embryo development.

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Sequence analysis, expression patterns and transcriptional regulation of mouse Ifrg15 during preimplantation embryonic development

Cited by 5 publications

References 30 publications

Oct4 regulates DNA methyltransferase 1 transcription by direct binding of the regulatory element

Oct4 regulates DNA methyltransferase 1 transcription by direct binding of the regulatory element

H3K27me3 may be associated with Oct4 and Sox2 in mouse preimplantation embryos

The interferon α-responsive gene, Ifrg15, plays vital roles during mouse early embryonic development

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