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Shuang Liang,Ming-Hui Zhao,Jung-Woo Kwon,Seon-Hyang Kim,Nam-Hyung Kim,Xiang-Shun Cui 한국동물생명공학회(구 한국동물번식학회) 2014 Reproductive & Developmental Biology(Supplement) Vol.38 No.2s
After somatic cell nuclear transfer (SCNT), epigenetic state of a differentiated donor cell nucleus must be reversed to the embryonic state. Incomplete epigenetic reprogramming is thought to be causing the low cloning efficiency. The present study was carried out to investigate the effects and mechanism of scriptaid, a novel histone deacetylase inhibitor (HDACi), on the in vitro development of porcine SCNT embryos. We found that treating SCNT embryos with 300 or 500 nM scriptaid for 20 h increased the development of embryos to the blastocyst stage and total cell numbers in per blastocyst( p<0.05). Apoptosis in blastocyst was decreased (p<0.05) in the presence of scriptaid. Scriptaid treatment significantly (p<0.05) increased the levels of H3-acK9 and 5-hmc, and the levels of H3-m3K9 and 5-mc were decreased at the pronuclear stage. Expression level of mir-152 was significantly (p<0.05) increased in scriptaid treated embryos. Moreover, treating embryos with 300 nM scriptaid increased the expression level of the genes that play important roles during embryonic development (OCT4 and CDX2). Additionally, mRNA expression of Dnmt1, Cas3 and Bak were significantly decreased and Bcl-xL was increased in scriptaid treated group compared to control. In conclusion, scriptaid improves the developmental capacity, prevent apoptosis through improving the epigenetic reprogramming in porcine SCNT embryos.
Down-Regulation of PP2A-B55α Impairs Mouse Preimplantation Embryo Development
Shuang Liang,Nam-Hyung Kim,Xiang-Shun Cui 한국수정란이식학회 2016 한국수정란이식학회 학술대회 Vol.2016 No.10
PP2A-B55α, a regulatory subunit of PP2A plays an important roles in regulating cell proliferation and survival. However, the functions for PP2A-B55α in mouse early embryo development is not clear. The objective of present were to investigate the expression patterns and to explore its biological function during mouse early development. Thetranscripts of PP2A-B55α were detected at all developmental stages in mouse embryo and decreased during embryo development. Immunostaining revealed that PP2A-B55α was present in both the nucleus and cytoplasm in early cleavage stage embryos. In the late embryonic development, PP2A-B55α was predominantly located in the cytoplasm. Knockdown (KD) of PP2A-B55α using double strand RNA not affect the proportion of cleaved embryos, but resulted in significantly decreased development to blastocyst stage and reduced total cell number in blastocyst. KD PP2A-B55α is able to induce sustained DNA damage and reduced the transcripts of non-homologous end joining (NHEJ) or homologous recombination (HR) pathways relative genes in mouse early embryo. KD PP2A-B55αcaused apoptosis and increase the transcripts of pro-apoptotic genes in blastocyst. Furthermore, The KDPP2A-B55α showed significantly lower cell proliferating rates (from 5-Bromo-deoxyuridineassayresults) in blastocysts and to talareas of out growth potential was decreased. These observation provide novel in sight into PP2A-B55α expression patterns in mouse early embryos and down-regulation of PP2A-B55α negatively impacted blastocyst development, total cell number, DNA damage, apoptosis, and proliferation and post-hatchingevents.
The Role of Melatonin on Mammalian Early Embryonic Development In Vitro
Shuang Liang,Xiang-Shun Cui 충북대학교 동물생명과학연구소 2016 동물생명과학연구 Vol.8 No.-
Melatonin is a hormone produced in the brain by the pineal gland from the amino acid tryptophan. It regulates a variety of important central and peripheral actions related to circadian rhythms and reproduction. Melatonin has a multifunctional molecule that mediates several circadian and seasonal processes in animal reproduction. Melatonin and its metabolites are antioxidants and free radical scavengers and a broad-spectrum antioxidant. In addition, it up-regulates gene expression and activity of several antioxidant enzymes including glutathione peroxidase and superoxide dismutase. Different from other classic antioxidants, its metabolites also exhibit antioxidant capacity. These successive actions are classified as the antioxidant cascade of melatonin. Melatonin also has been found to preserve optimal mitochondrial function and homeostasis by protecting against mitochondrial oxidative stress, thereby curtailing subsequent apoptotic events and cell death. High concentrations of melatonin have been found in mammalian preovulatory follicular fluids. Several studies indicated that melatonin can improve mammalian embryo development. In this review, we summarize the effect of melatonin on mammalian oocytes maturation and early embryo development.
Liang, Shuang,Nie, Zheng-Wen,Guo, Jing,Niu, Ying-Jie,Shin, Kyung-Tae,Ock, Sun A.,Cui, Xiang-Shun Cambridge University Press 2018 Microscopy and Microanalysis Vol.24 No.1
<B>Abstract</B><P>MicroRNA (miR)-29b plays a crucial role during somatic cell reprogramming. The aim of the current study was to explore the effects of miR-29b on the developmental competence of bovine somatic cell nuclear transfer (SCNT) embryos, as well as the underlying mechanisms of action. The expression level of miR-29b was lower in bovine SCNT embryos at the pronuclear, 8-cell, and blastocyst stages compared with <I>in vitro</I> fertilized embryos. In addition, miR-29b regulates the expression of DNA methyltransferases (<I>Dnmt3a/3b</I> and <I>Dnmt1</I>) in bovine SCNT embryos. We further investigated SCNT embryo developmental competence and found that miR-29b overexpression during bovine SCNT embryonic development does not improve developmental potency and downregulation inhibits developmental potency. Nevertheless, the quality of bovine SCNT embryos at the blastocyst stage improved significantly. The expression of pluripotency factors and cellular proliferation were significantly higher in blastocysts from the miR-29b overexpression group than the control and downregulation groups. In addition, outgrowth potential in blastocysts after miR-29b overexpression was also significantly greater in the miR-29b overexpression group than in the control and downregulation groups. Taken together, these results demonstrated that miR-29b plays an important role in bovine SCNT embryo development.</P>
Liang, Shuang,Guo, Jing,Choi, Jeong-Woo,Kim, Nam-Hyung,Cui, Xiang-Shun CSIRO Publishing 2017 Reproduction, fertility, and development Vol.29 No.9
<P> After reaching the metaphase II (MII) stage, unfertilised oocytes undergo a time-dependent process of quality deterioration referred to as oocyte aging. The associated morphological and cellular changes lead to decreased oocyte developmental potential. This study investigated the effect of exogenous melatonin supplementation on in vitro aged bovine oocytes and explored its underlying mechanisms. The levels of cytoplasmic reactive oxygen species and DNA damage response in bovine oocytes increased during in vitro aging. Meanwhile, maturation promoting factor activity significantly decreased and the proportion of morphologically abnormal oocytes significantly increased. Melatonin supplementation significantly decreased quality deterioration in aged bovine MII oocytes (P < 0.05). Additionally, it decreased the frequency of aberrant spindle organisation and cortical granule release during oocyte aging (P < 0.05). In the melatonin-supplemented group, mitochondrial membrane potential and ATP production were significantly increased compared with control. Furthermore, melatonin treatment significantly increased the speed of development of bovine oocytes to the blastocyst stage after in vitro fertilisation and significantly decreased the apoptotic rate in the blastocysts (P < 0.05). The expression of Bax and Casp3 in the blastocysts was significantly reduced after treatment with melatonin, whereas expression of Bcl2 significantly increased (P < 0.05). In conclusion, these findings suggest that supplementation of aged bovine oocytes with exogenous melatonin improves oocyte quality, thereby enhancing the developmental capacity of early embryos. </P>
Protective Effects of Coenzyme Q10 on Developmental Competence of Porcine Early Embryos
Liang, Shuang,Niu, Ying Jie,Shin, Kyung-Tae,Cui, Xiang-Shun Cambridge University Press 2017 Microscopy and Microanalysis Vol.23 No.4
<B>Abstract</B><P>Coenzyme Q10 (Q10) plays an important role in the cellular antioxidant system by protecting the cells from free-radical oxidative damage and apoptosis. In the present study, we have investigated the effect of Q10 on the preimplantation development of porcine parthenogenetic embryos, as well as the underlying mechanism. The results showed that 100 <I>μ</I>M was the optimal concentration of Q10, which resulted in significantly increased cleavage and blastocyst formation rates and improvement of blastocyst quality. Q10 improved the blastocyst hatching rate and cellular proliferation rate in hatching blastocysts and increased the expression of hatching-related genes. Furthermore, Q10 not only decreased reactive oxygen species production, DNA damage levels, and apoptosis in the blastocysts from H2O2-induced oxidative injury, but also maintained mitochondrial function. Taken together, these results indicate that Q10 has beneficial effects on the development of porcine parthenogenetic embryos by preventing oxidative damage and apoptosis.</P>
Glucose Metabolism Influences Cytoplasmic Maturation in Porcine Oocytes
Bao Yuan,Shuang Liang,Jiabao Zhang,Jeong-Woo Kwon,Shun-Ha Park,Hai-Yang Wang,PIAO XUAN JING,Xiang-Shun Cui,Nam-Hyung Kim 한국수정란이식학회 2016 한국수정란이식학회 학술대회 Vol.2016 No.10
In the present study, we examined potential roles of glucose and pyruvate in nuclear and cytoplasmic maturation of porcine oocytes. In the presence and absence of 10% porcine follicular fluid (PFF), either 5.6 mM glucose or 2mM pyruvate effect on meiotic maturation and followed development ability. However, DOs doesn't take full advantage of the glucose in medium, only pyruvate can increase MII rate and follow early embryo development ability significance. COCs were matured with 200 uM pentose phosphate pathway (PPP) inhibitor (dehydroepiandrosterone, DHEA) or 2 μM glycolysis inhibitor (iodoacetate, IA), significantly lower levels of GHS in the DHEA an IA treated oocytes and the levels of ROS were higher significantly in the DHEA treated oocytes, treatment with DHEA significantly reduced the intra-oocyte ATP and NADPH level. Blastocysts from DHEA or IA treated group also presented higher apoptosis levels, meanwhile, the percentage of proliferating cells was dramatically lower than the non-treated group. In conclusion, our results suggest that 10% PFF promoted oocytes make full use of energy, glucose metabolism during in vitro maturation inseparable from the cumulus cells, PPP and glycolysis promoted porcine oocytes cytoplasmic maturation by supplying energy and reducing oxidative stress.