Treatment of fetal fibroblasts with DNA methylation inhibitors and/or histone deacetylase inhibitors improves the development of porcine nuclear transfer-derived embryos

Diao, Yun Fei; Naruse, Kenji; Han, Rong Xun; Li, Xiaoxia; Oqani, Reza K.; Lin, Tao; Jin, Dong Il

doi:10.1016/j.anireprosci.2013.08.008

Cited by 29 publications

(30 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the present study, we first investigated the effects of Zb treatment in a porcine IVP system, and we found that supplementation with Zb during IVC could affect the subsequent embryo development of SCNT embryos in different ways according to its concentration and treatment time. The results in this study are consistent with a previous study by Diao et al (), in which a low dose of Zb (5 nM) significantly improved the SCNT blastocyst development rate. Interestingly, although there was no detrimental effect on the percentage of cleaved embryos in a dose‐dependent manner, blastocyst development was significantly impaired when SCNT embryos were treated with a high dose of Zb (50 nM) compared to the low dose treatment groups (5 and 10 nM).…”

Section: Discussionsupporting

confidence: 93%

Enhancement of epigenetic reprogramming status of porcine cloned embryos with zebularine, a DNA methyltransferase inhibitor

Taweechaipaisankul

Jin

Lee

et al. 2019

Molecular Reproduction Devel

View full text Add to dashboard Cite

Aberrant epigenetic reprogramming is known to be a major cause of inefficient somatic cell nuclear transfer (SCNT) in pigs, and use of epigenetic modification agents, such as DNA methyltransferase inhibitors (DNMTis), is a promising approach for enhancing SCNT efficacy. Here, we attempted to find the optimal condition of zebularine (Zb), a DNMTi, treatment on porcine SCNT embryos during in vitro culture (IVC). As results, treatment with 5 nM Zb for 24 hr showed the highest rate of embryo development to blastocyst compared to other groups (p < .05). Also, the relative intensities of global DNA methylation levels of anti‐5‐methylcytosine in pseudo‐pronuclear (PNC), 2‐cell and 4‐cell stages were significantly lower in the Zb‐treated group (p < .05), however, changes in methylation levels of centromeric satellite repeat were noted only in PNC and blastocyst stages. In addition, significant positive alterations in the relative expression of genes related to pluripotency (OCT4 and SOX2), histone acetylation (HAT1, HDAC1, HDAC2, and HDAC3) and DNA methylation (DNMT1 and DNMT3a) were observed compared to the control (p < .05). In conclusion, we found that Zb could modify DNA methylation levels in the early stages of porcine SCNT embryos and promote their developmental competence.

show abstract

Section: Discussionsupporting

confidence: 93%

Enhancement of epigenetic reprogramming status of porcine cloned embryos with zebularine, a DNA methyltransferase inhibitor

Taweechaipaisankul

Jin

Lee

et al. 2019

Molecular Reproduction Devel

View full text Add to dashboard Cite

show abstract

“…[35]. Cumulus-free oocytes in PZM-3 medium containing 7.5 µg/ml cytochalasin B at 38°C were enucleated by aspirating the first polar body and adjacent cytoplasm with a fine glass pipette.…”

Section: Methodsmentioning

confidence: 99%

Changes in Histone H3 Lysine 36 Methylation in Porcine Oocytes and Preimplantation Embryos

Diao

Oqani

et al. 2014

PLoS ONE

Self Cite

View full text Add to dashboard Cite

Histone H3 lysine 36 (H3K36) methylation is known to be associated with transcriptionally active genes, and is considered a genomic marker of active loci. To investigate the changes in H3K36 methylation in pig, we determined the mono-, di-, and tri-methylations of H3K36 (H3K36me1, H3K36me2 and H3K36me3, respectively) in porcine fetal fibroblasts, oocytes and preimplantation embryos by immunocytochemistry using specific antibodies and confocal microscopy. These analyses revealed that only H3K36me3 in porcine fetal fibroblasts consistently colocalized with transcription sites identified as actively synthesizing RNA based on fluorouridine (FU) incorporation. Treatment of cells with flavopiridol, which blocks transcription elongation, completely abrogated both H3K36me3 signals and RNA synthesis. All three types of H3K36 methylation were present and did not significantly differ during oocyte maturation. In parthenogenetic embryos, H3K36me1 and -me2 were detected in 1-cell through blastocyst-stage embryos. In contrast, H3K36me3 was not detected in most 1-cell stage embryos. H3K36me3 signals became detectable in 2-cell stage embryos, peaked at the 4-cell stage, decreased at the 8-cell stage, and then became undetectable at blastocyst stages in both parthenogenetic and in vitro-fertilized (IVF) embryos. Unlike the case in IVF embryos, H3K36me3 could not be demethylated completely during the 1-cell stage in somatic cell nuclear transfer (SCNT) embryos. These results collectively indicate that H3K36me3, but not H3K36me1 or -me2, is associated with transcription elongation in porcine fetal fibroblasts. H3K36me3 is developmentally regulated and may be a histone mark of embryonic gene activation in pig. Aberrant H3K36 tri-methylation occurred during the nuclear reprogramming of SCNT embryos.

show abstract

“…RG108, was found to be free of cytotoxic or genotoxic effects compared to other DNMTis, such as 5-aza-2-deoxycytidine (5-aza-dC) [12], zebularine [13], and epigallocatechin-3-gallate [14], and had been used to assist the somatic nucleus to mimic DNA methylation and chromatin remodeling. In pigs, RG108 treatment improved the developmental capacity of cloned embryos [15]. However, research into these potential applications progresses slowly in pigs, because the cloning efficiency is extremely low and no authentic porcine embryonic stem cells are currently available [16][17][18][19].…”

Section: Introductionmentioning

confidence: 99%

Dynamic Methylation Changes of DNA and H3K4 by RG108 Improve Epigenetic Reprogramming of Somatic Cell Nuclear Transfer Embryos in Pigs

Zhai

Zhang

et al. 2018

Cell Physiol Biochem

View full text Add to dashboard Cite

Background/Aims: DNA methylation and histone modifications are essential epigenetic marks that can significantly affect the mammalian somatic cell nuclear transfer (SCNT) embryo development. However, the mechanisms by which the DNA methylation affects the epigenetic reprogramming have not been fully elucidated. Methods: In our study, we used quantitative polymerase chain reaction (qPCR), Western blotting, immunofluorescence staining (IF) and sodium bisulfite genomic sequencing to examine the effects of RG108, a DNA methyltransferase inhibitor (DNMTi), on the dynamic pattern of DNA methylation and histone modifications in porcine SCNT embryos and investigate the mechanism by which the epigenome status of donor cells’ affects SCNT embryos development and the crosstalk between epigenetic signals. Results: Our results showed that active DNA demethylation was enhanced by the significantly improving expression levels of TET1, TET2, TET3 and 5hmC, and passive DNA demethylation was promoted by the remarkably inhibitory expression levels of DNMT1, DNMT3A and 5mC in embryos constructed from the fetal fibroblasts (FFs) treated with RG108 (RG-SCNT embryos) compared to the levels in embryos from control FFs (FF-SCNT embryos). The signal intensity of histone H3 lysine 4 trimethylation (H3K4me3) and histone H3 lysine 9 acetylation (H3K9Ac) was significantly increased and the expression levels of H3K4 methyltransferases were more than 2-fold higher expression in RG-SCNT embryos. RG-SCNT embryos had significantly higher cleavage and blastocyst rates (69.3±1.4%, and 24.72±2.3%, respectively) than FF-SCNT embryos (60.1±2.4% and 18.38±1.9%, respectively). Conclusion: Dynamic changes in DNA methylation caused by RG108 result in dynamic alterations in the patterns of H3K4me3, H3K9Ac and histone H3 lysine 9 trimethylation (H3K9me3), which leads to the activation of embryonic genome and epigenetic modification enzymes associated with H3K4 methylation, and contributes to reconstructing normal epigenetic modifications and improving the developmental efficiency of porcine SCNT embryos.

show abstract

Treatment of fetal fibroblasts with DNA methylation inhibitors and/or histone deacetylase inhibitors improves the development of porcine nuclear transfer-derived embryos

Cited by 29 publications

References 30 publications

Enhancement of epigenetic reprogramming status of porcine cloned embryos with zebularine, a DNA methyltransferase inhibitor

Enhancement of epigenetic reprogramming status of porcine cloned embryos with zebularine, a DNA methyltransferase inhibitor

Changes in Histone H3 Lysine 36 Methylation in Porcine Oocytes and Preimplantation Embryos

Dynamic Methylation Changes of DNA and H3K4 by RG108 Improve Epigenetic Reprogramming of Somatic Cell Nuclear Transfer Embryos in Pigs

Contact Info

Product

Resources

About