Ascorbic acid improves parthenogenetic embryo development through TET proteins in mice

Yu, Xiaofeng; Hao, Jindong; Qi, Ming‐Hui; Liang, Han; Lin, Chao; Wang, Dongxu

doi:10.1042/bsr20181730

Cited by 11 publications

(10 citation statements)

References 29 publications

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“…Immunofluorescence (IF) staining is an important method for measuring protein expression in embryos, and has been used in our previous studies (Gao et al, 2019; Hao et al, 2019; Liang et al, 2017; Yuan et al, 2017) and other related studies (Amouroux et al, 2016; Inoue & Zhang, 2011; Sakashita et al, 2014). To detect expression of apoptotic proteins in MII oocytes by IF staining in this study, we analyzed the P53 level of 21 MII oocytes of 100 μM PB1 group and 21 MII oocytes of control group, the caspase‐3 level of 24 MII oocytes of 100 μM PB1 group and 23 MII oocytes of control group.…”

Section: Methodsmentioning

confidence: 99%

Procyanidin B1 promotes in vitro maturation of pig oocytes by reducing oxidative stress

Jin

Hao

Huang

et al. 2020

Molecular Reproduction Devel

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Oxidative stress negatively affects the in vitro maturation (IVM) of oocytes. Procyanidin B1 (PB1) is a natural polyphenolic compound that has antioxidant properties. In this study, we investigated the effect of PB1 supplementation during IVM of porcine oocytes. Treatment with 100 μM PB1 significantly increased the MII oocytes rate (p <0.05), the parthenogenetic (PA) blastocyst rate (p <0.01) and the total cell number in the PA blastocyst (p < 0.01) which were cultured in regular in vitro culture (IVC) medium. The PA blastocyst rate of regular MII oocytes activated and cultured in IVC medium supplemented with 100 and 150 μM PB1 significantly increased compared with control (p < 0.01 and p < 0.05). We also evaluated the reactive oxygen species (ROS) levels, mitochondrial membrane potential (Δψm) levels, glutathione (GSH) levels, and apoptotic levels in MII oocytes and cumulus cells following 100 μM PB1 treatment. The results showed that the PB1 supplementation decreased ROS production and apoptotic levels. In addition, PB1 was found to increase Δψm levels and GSH levels. In conclusion, PB1 inhibited apoptosis of oocytes and cumulus cells by reducing oxidative stress. Moreover, PB1 improved the quality of oocytes and promoted PA embryo development. Taken together, our results suggest that PB1 is a promising antioxidant additive for IVM of oocytes.

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Section: Methodsmentioning

confidence: 99%

Procyanidin B1 promotes in vitro maturation of pig oocytes by reducing oxidative stress

Jin

Hao

Huang

et al. 2020

Molecular Reproduction Devel

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“…Several epigenetic modifications, such as DNA methylation and histone modification, have been observed to be lacked during the development of parthenogenetic embryos because of the deficiency of HDAC1 and DNMT1 expression [48]. In addition, downregulated expression of TET1, TET2, and TET3 has been confirmed in parthenogenetic embryos [49]. To detect CENPF function in both IVF and parthenogenetic embryos, MII oocytes and zygotes were used for CENPF knockdown.…”

Section: Discussionmentioning

confidence: 99%

Loss of CENPF leads to developmental failure in mouse embryos

et al. 2019

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Aneuploidy caused by abnormal chromosome segregation during early embryo development leads to embryonic death or congenital malformation. Centromere protein F (CENPF) is a member of centromere protein family that regulates chromosome segregation during mitosis. However, its necessity in early embryo development has not been fully investigated. In this study, expression and function of CENPF was investigated in mouse early embryogenesis. Detection of CENPF expression and localization revealed a cytoplasm, spindle and nuclear membrane related dynamic pattern throughout mitotic progression. Farnesyltransferase inhibitor (FTI) was employed to inhibit CENPF farnesylation in zygotes. The results showed that CENPF degradation was inhibited and its specific localization on nuclear membranes in morula and blastocyst vanished after FTI treatment. Also, CAAX motif mutation leads to failure of CENPF-C630 localization in morula and blastocyst. These results indicate that farnesylation plays a key role during CENPF degradation and localization in early embryos. To further assess CENPF function in parthenogenetic or fertilized embryos development, morpholino (MO) and Trim-Away were used to disturb CENPF function. CENPF knockdown in Metaphase II (MII) oocytes, zygotes or embryos with MO approach resulted in failure to develop into morulae and blastocysts, revealing its indispensable role in both parthenogenetic and fertilized embryos. Disturbing of CENPF with Trim-Away approach in zygotes resulted in impaired development of 2-cell and 4-cell, but did not affect the morula and blastocyst formation because of the recovered expression of CENPF. Taken together, our data suggest CENPF plays an important role during early embryonic development in mice.

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“…AA, as an irreplaceable cofactor, can effectively enhance the activity of the TET3 enzyme and promote the demethylation of active DNA (Gao et al, ). A previous study showed that in sheep IVF preimplantation embryos, the transcriptional level of TET3 was highest in the 2‐cell stage and gradually decreased in the 4‐cell, 8‐cell, and morula stages until the blastocyst stage, where it was the lowest.…”

Section: Discussionmentioning

confidence: 99%

“…In this study, TET3 fluorescence intensity analysis showed that TET3 gradually increased from the 2cell to the morula stage and was weakly present in the blastocyst stage, unlike the results of the study in cattle, TET3 is expressed in the cytoplasm at all stages of IVF preimplantation embryo development in the yak. Perhaps the previous results can explain this(Yuko et al, 2012), first, there is a highly expressed activation-induced cytidine deaminase (Aid) in the development of yak embryos, and coexpression pattern with TET, secondly, the shuttle function of Aid transfers nuclear TET3 protein to the cytoplasm, and in this study the gene expression trend of TET3 was consistent with the fluorescence intensity analysis results, which indicates that the TET3 protein regulates the mechanism of active DNA demethylation by subcellular localization.AA, as an irreplaceable cofactor, can effectively enhance the activity of the TET3 enzyme and promote the demethylation of active DNA(Gao et al, 2019). A previous study showed that in sheep IVF preimplantation embryos, the transcriptional level of TET3 was highest in the 2-cell stage and gradually decreased in the 4-cell, 8-cell, and morula stages until the blastocyst stage, where it was the lowest.…”

mentioning

confidence: 99%

DNA methylation regulated by ascorbic acids in yak preimplantation embryo helps to improve blastocyst quality

et al. 2019

Molecular Reproduction Devel

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DNA methylation as an important, essential epigenetic modification is critical for the successful development of mammalian embryos. In recent years, the important role of ascorbic acid (AA) as an irreplaceable cofactor for epigenetic regulation has been confirmed. However, the effect of AA on DNA methylation in preimplantation embryo development of plateau yak remains unknown. In this study, we explored whether AA can help regulates DNA methylation in yak preimplantation embryos to improve the blastocyst quality. First, our results indicate that the preimplantation of the yak still follows the classical pattern of DNA demethylation and remethylation, however, remethylation occurs in the blastocyst stage. Second, the unique expression pattern of the ten‐eleven translocation enzyme (TET3) in the cytoplasm plays a key role in the demethylation mechanism. Third, in the blastocyst stage, the pluripotency gene CDX2 promoter region was in a hypomethylated state, and the POU5F1, SOX2, and NANOG promoter regions were in moderate methylation states. In addition, treatment with 50 μg/ml AA mainly improved the expression levels of DNMT1, DNMT3a, and TET3, ensured the establishment, maintenance and transition of 5‐methylcytosine. After AA treatment, the methylation level of the pluripotency genes NANOG promoter regions was significantly reduced, and the mRNA transcript abundance of the pluripotency genes NANOG, POU5F1, and CDX2 was upregulated. In conclusion, our findings suggest that AA could increase blastocyst cell numbers by regulating DNA methylation of yak preimplantation embryos .

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Ascorbic acid improves parthenogenetic embryo development through TET proteins in mice

Cited by 11 publications

References 29 publications

Procyanidin B1 promotes in vitro maturation of pig oocytes by reducing oxidative stress

Procyanidin B1 promotes in vitro maturation of pig oocytes by reducing oxidative stress

Loss of CENPF leads to developmental failure in mouse embryos

DNA methylation regulated by ascorbic acids in yak preimplantation embryo helps to improve blastocyst quality

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