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Epigenetic reprogramming in nuclear transferred embryos Autophagy and mitochondrial aspect
XingHui Shen 한국동물생명공학회(구 한국동물번식학회) 2017 발생공학 국제심포지엄 및 학술대회 Vol.2017 No.10
Autophagy is an essential cellular mechanism that degrades cytoplasmic proteins and organelles to recycle their components. Moreover, autophagy is essential for preimplantation development in mammals. Here we show that autophagy is also important for reprogramming in somatic cell nuclear transfer (SCNT). Our data indicate that unlike fertilized oocytes, autophagy is not triggered in SCNT embryos during 6 hours of activation. Mechanistically, the inhibited autophagic induction during SCNT activation is due to the cytochalasin B (CB) caused depolymerization of actin filaments. In this study, we induced autophagy during SCNT activation by rapamycin and pp242, which could restore the expected level of autophagy and significantly enhance the development of SCNT embryos to the blastocyst stage when compared with the control (68.5% and 68.7% vs. 41.5%, p<0.05). Furthermore, the treatment of rapamycin and pp242 accelerates active DNA demethylation indicated by the conversion of 5 mC to 5 hmC, and treatment of rapamycin improves degradation of maternal mRNA as well. Thus, our findings reveal that autophagy is important for development of SCNT embryos and inhibited autophagic induction during SCNT activation might be one of the serious causes of low efficiency of SCNT.
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2i System Enhances Porcine Blastocyst Quality by Regulating of Epigenetic Modification
Jeongwoo Kwon,XingHui Shen,Jiang Hao,Yao Xuerui,Nam-Hyung Kim 한국동물생명공학회(구 한국동물번식학회) 2017 발생공학 국제심포지엄 및 학술대회 Vol.2017 No.10
2i system is useful for maintenance of navie state of embryonic stem cells in various species. However, the role of 2i system in porcine preimplantation stage embryos unclear. In this study, we investigated the quality of blastocyst and pluripotent-related factor by 2i system. And, we also investigated about DNA and RNA epigenetic modification status by 2i system during porcine early embryo development. After treatment of MEK1/2 inhibitor PD0325901(4 μM), GSK-3 inhibitor CHIR99021(0.3 μM) and 2i (PD0325901+CHIR99021) during pre-implantation development, blastocyst formation rate had no significantly difference. However, blastocyst size increased in 2i treatment embryos. Blastocyst quality also increased after treatment of CHIR and 2i using TUNEL assay and Brdu assay. Interestingly, main transcription factor OCT4 and SOX2 positive cells in blastocyst was significantly increased in 2i groups. Especially, histone modification- relation proteins like h3k9me3 and h3k9ac changed after treatment of CHIR and 2i. Global gene expression patterns related with pluripotency and epigenetic also changed in inhibitor treatment groups. Furthermore, bi-sulfite sequencing results showed that 2i treatment groups demethylated in satellite I region. In conclusion, our findings strongly suggest that 2i system can be useful method for increasing of blastocyst quality by affecting of ICM/TE formation by regulating epigenetic modification during early embryo development.
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Inhibition of MEK1/2 and GSK-3 Enhance Quality of Porcine Blastocyst Culturing In Vitro
Jeongwoo Kwon,Yujin Jo,XingHui Shen,Nam-Hyung Kim 한국동물생명공학회(구 한국동물번식학회) 2017 Reproductive & Developmental Biology(Supplement) Vol.41 No.2
Inhibition of MEK1/2 and GSK-3(2i system) is useful for maintenance of navie state of embryonic stem cells in various species. However, the role of 2i system in porcine preimplantation stage embryos is not clear. In this study, we investigated the quality of blastocyst and pluripotent-related factors by 2i system and epigenetic modification status by 2i system during porcine early embryo development. Treatment of MEK1/2 inhibitor PD0325901 (4 uM), GSK-3 inhibitor CHIR99021 (0.3 uM) and 2i (PD0325901+ CHIR99021) did not enhanced the rate of blastocyst formation. However, the size of blastocyst was increased in 2i treated embryos. TUNEL assay showed that CHIR- 99021 and 2i treatment enhanced blastocyst quality. Furthermore, 2i treated blastocyst increased OCT4 and SOX2 positive cells and decreased GATA4 positive cells. Methylation of histone H3 lysin trime3 (h3k9me3) were reduced, and histone H3 deacetylaton (h3k9ac) were increased in 2i treated embryos. Furthermore, bi-sulfite sequencing results showed that 2i treatment groups demethylated in satellite I region. And, RNA methylatransferase protein METTL3 was increased while demethylase FTO was increased after treatment of 2i. In conclusion, our findings strongly suggest that 2i system can be useful method for increasing of blastocyst quality by affecting of ICM/TE formation by regulating epigenetic modification during early embryo development.
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Laminarin enhances the quality of aged pig oocytes by reducing oxidative stress
YAO, Xuerui,JIANG, Hao,LIANG, Shuang,SHEN, Xinghui,GAO, Qingshan,XU, Yong Nan,KIM, Nam-Hyung 家畜繁殖硏究所 2018 Journal of Reproduction and Development Vol.64 No.6
<P>Laminarin (LAM) is a β-glucan oligomer known to possess biological activities such as anticancer and antioxidant effects. This study explored the influence of LAM supplementation on <I>in vitro</I> aged porcine oocytes and the underlying mechanisms behind this influence. We found that LAM delayed the aging process and improved the quality of aged oocytes. LAM supplementation enhanced the subsequent developmental competence of aged oocytes during the <I>in vitro</I> aging process. The blastocyst formation rate was significantly increased in aged oocytes treated with 20 µg/ml LAM compared to non-treated aged oocytes (45.3% <I>vs</I>. 28.7%, P < 0.01). The mRNA levels of apoptosis-related genes, B cell lymphoma-2-associated X protein (<I>Bax)</I> and <I>Caspase-3,</I> were significantly lower in blastocysts derived from the LAM-treated aged oocytes during the <I>in vitro</I> aging process. Furthermore, the level of intracellular reactive oxygen species was significantly decreased and that of glutathione was significantly increased in aged oocytes following LAM treatment. Mitochondrial membrane potential was increased, and the activities of caspase-3 and cathepsin B were significantly reduced in the LAM-treated aged oocytes compared with the non-treated aged oocytes. Taken together, these results suggest that LAM is beneficial for delaying the aging process in porcine oocytes.</P>
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Yuanfeng Wu,Xianghai Song,Shuai Li,Jiahui Zhang,Xinghui Yang,Pengxin Shen,Lijing Gao,Ruiping Wei,Jin Zhang,Guomin Xiao 한국공업화학회 2018 Journal of Industrial and Engineering Chemistry Vol.58 No.-
[(CH3)2NH2][M3(BTC)(HCOO)4(H2O)].H2O (M–BTC, M = Mn, Ni, Co) were prepared under hydrothermal conditions and used as highly efficient catalysts for cycloaddition of CO2 with epichlorohydrin (ECH). The microstructure and physicochemical properties of the compounds were determined by PXRD, FT-IR, XPS, N2-adsorption, TG–DSC, NH3–TPD and CO2–TPD. 98.01% conversion of ECH and 96.05% selectivity to chloropropene carbonate was obtained over the Mn–BTC under the optimized reaction conditions (105 °C, 3.0 MPa, 9 h, 1.5 wt.% of ECH). Besides, the recyclability result exhibited the Mn–BTC compound can be utilized as least three times with a slight reduction in its catalytic ability. In addition, cycloaddition of CO2 with other epoxides and DFT calculation were also performed. The result exhibited the yield followed the order: ECH > 1, 2-epoxybutane > propene oxide > Allyl glycidyl ether, which was mainly determined by the energy of reaction.
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