Effect of supplemetation of Zebularine and Scriptaid on efficiency of <i>in vitro</i> developmental competence of ovine somatic cell nuclear transferred embryos

Fang, Xiaohuan; Xia, Wei; Cao, Hui; Guo, Yanhua; Wang, Han; Zhang, Xiaosheng; Wan, Pengcheng; Qiaoli, Wei; Sun, Song; Tian, Shujun; Liu, Jun-Jie; Wang, Zhigang

doi:10.1080/10495398.2018.1559846

Cited by 11 publications

(7 citation statements)

References 41 publications

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“…These data suggest that BCB staining highlights the most competent oocytes, irrespective of follicular size. Our results are consistent with those of other studies, where BCB+ oocytes had a reduced G6PDH activity, and where oocytes from large follicles had probably completed their growth (DE BEM et al, 2014;FANG et al, 2019;KIM et al, 2010).…”

Section: Discussionsupporting

confidence: 93%

“…We discovered that the follicular size of oocytes affects cleavage and blastocyst rates, regardless of their BCB classification, whereas the proportion of hatched blastocysts and the mean number of embryonic cells did not differ between treatments. These results are partly consistent with those of other studies, which suggest that oocytes from larger follicles are more competent (DE BEM et al, 2014), and that BCB+ oocytes result in higher blastocyst rates as compared to BCB-oocytes (FANG et al, 2019).…”

Section: Discussionsupporting

confidence: 92%

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In vitro embryos production of oocytes selected based on follicular size and by brilliant cresyl blue staining

Barros

Adona

Guemra

et al. 2019

SCA

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We evaluated the competence of bovine oocytes via morphological selection associated with follicular diameter and by brilliant cresyl blue (BCB) staining, in order to improve oocyte selection and the efficiency of in vitro embryo production. Follicles with small (2 < 4 mm), large (4–8 mm) or mixed (2–8 mm) diameters were aspirated from slaughterhouse ovaries. Next, the morphologically categorized oocytes were stained and evaluated with 26 ?M of BCB (90 min), followed by an assessment of in vitro maturation (MII) and blastocyst production. Oocytes from the large (61.1 ± 3.9%) and mixed (52.9 ± 4.6%) follicular groups showed comparable (p > 0,05) BCB+ rates, but both differed (p < 0.05) as compared to the small-diameter group (41.0 ± 4.5%). Between the BCB+ and BCB – oocytes, there was no difference (p > 0,05) in MII rate in the mixed (respectively 86.7 ± 3.4 and 75.8 ± 2.2), large (respectively 89.4 ± 2.6 and 79.7 ± 2.3) and small (respectively 76.7 ± 2.9 and 67.2 ± 2.5) diameter groups. Comparing BCB (+ and -) oocytes, we observed (p > 0,05) no change in the percentage of embryos in the large group (respectively 67.3 ± 3.3 and 53.5 ± 5.8) and in the small group (respectively 43.8 ± 5.3 and 33.9 ± 2.2), but there was a difference (p < 0.05) in the mixed group (respectively 57.8 ± 3.9 and 39.6 ± 7.5). Furthermore, when we compared between follicular groups, the BCB+/large-diameter group (61.1 ± 3.9%) had a significant higher percentage of embryos (p < 0.05) than the small-diameter BCB+ (43.8 ± 5.3) groups. To conclude, selection procedures that use follicle size and/or BCB staining may be used as an additional tool to improve the in vitro production of embryos.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionsupporting

confidence: 92%

In vitro embryos production of oocytes selected based on follicular size and by brilliant cresyl blue staining

Barros

Adona

Guemra

et al. 2019

SCA

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“…In most cancer cells, Zb shows a consistent pattern of growth inhibition and demethylation effects with little evidence of toxicity (Cheng et al, ; Cheng et al, ). In embryo production studies, Zb has been applied to modify the epigenetic status of donor fibroblast cells, and it could enhance the subsequent in vitro developmental competence and quality of SCNT embryos of pigs (Diao et al, ), yaks (Xiong et al, ), and sheep (Cao et al, ; Fang et al, ). 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.…”

Section: Discussionmentioning

confidence: 99%

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

Taweechaipaisankul

Jin

Lee

et al. 2019

Molecular Reproduction Devel

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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.

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“…In the future it would be interesting to focus on the DNA methylation levels of the selected genes (e.g., SOD1, SOD2) as well as on the genome-wide methylation pattern and the relationships with the gene expression. et al, 2018), but also by somatic cell cloning (Fang et al, 2020;Gorczyca et al, 2021;Nadri et al, 2022;Wiater et al, 2021).…”

Section: Discussionmentioning

confidence: 99%

Global DNA methylation, DNA methyltransferase and stress‐related gene expression in ovine oocytes and embryos after exposure to metabolic stressors

Nandi

Tripathi

Singh

et al. 2023

Reprod Domestic Animals

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DNA methylation, considered the most prominent epigenetic mark was important for the gene regulation in embryonic development. The present study aimed at evaluating the effects of metabolic stressors [Non‐esterified fatty acid (NEFA), β‐hydroxy‐butyric acid (BHB), ammonia and urea] exposure during the in vitro ovine oocyte maturation, global DNA methylation, DNA methyltransferase and stress‐related gene expression. Colorimetric analysis of global DNA methylation and the expression of the DNA methyltransferase genes (DNMT1, DNMT3A, and DNMT3B) were assessed in the matured oocytes, 2‐cell embryos and blastocysts produced in vitro from oocytes exposed with the metabolic stressors during 24 h of the in vitro maturation (IVM). Further, the mRNA expression of the stress‐related genes (SOD1, SOD2) in the matured oocytes, 2‐cell embryos and blastocysts produced was assessed. Significant difference in global DNA methylation levels between all the treatments tested was observed when compared with control in oocytes, two‐cell embryos and blastocysts. Elevated concentration of metabolic stressors resulted in increased expressions of several stress‐related genes, i.e., SOD1, SOD2 and in mRNA expression of DNA methyltransferase genes. The present study is the first to report that the DNA methylation was sensitive to the effects of the metabolic stressors in ovine oocytes/embryos. The aberrant expressions of genes during oocyte development targeted in the present study can provide evidence for the early embryo developmental arrest and blastocysts quality. These results highlighted the sensitivity of the early embryogenesis and more precisely of the reprogramming period to metabolites challenges, in a realistic situation of elevated concentration of metabolic stressors.

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Effect of supplemetation of Zebularine and Scriptaid on efficiency of in vitro developmental competence of ovine somatic cell nuclear transferred embryos

Cited by 11 publications

References 41 publications

In vitro embryos production of oocytes selected based on follicular size and by brilliant cresyl blue staining

In vitro embryos production of oocytes selected based on follicular size and by brilliant cresyl blue staining

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

Global DNA methylation, DNA methyltransferase and stress‐related gene expression in ovine oocytes and embryos after exposure to metabolic stressors

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