Ascorbic Acid Increases Demethylation in Somatic Cell Nuclear Transfer Embryos of the Pig ()

Overview

Journal Asian-Australas J Anim Sci

Specialty Veterinary Medicine

Date 2017 Jan 24

PMID 28111439

Citations 3

Authors

Minghui Zhao

Tai-Young Hur

Jingu No

Yoonseok Nam

Hyeunkyu Kim

Gi-Sun Im

Seunghoon Lee

Affiliations

Soon will be listed here.

Abstract

Objective: Investigated the effect and mechanism of ascorbic acid on the development of porcine embryos produced by somatic cell nuclear transfer (SCNT).

Methods: Porcine embryos were produced by SCNT and cultured in the presence or absence of ascorbic acid. Ten-eleven translocation 3 (TET3) in oocytes was knocked down by siRNA injection. After ascorbic acid treatment, reprogramming genes were analyzed by realtime reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, relative 5-methylcytosine and 5-hydroxymethylcytosine content in pronucleus were detected by realtime PCR.

Results: Ascorbic acid significantly increased the development of porcine embryos produced by SCNT. After SCNT, transcript levels of reprogramming genes, , , and were significantly increased in blastocysts. Furthermore, ascorbic acid reduced 5-methylcytosine content in pronuclear embryos compared with the control group. Knock down of TET3 in porcine oocytes significantly prevents the demethylation of somatic cell nucleus after SCNT, even if in the presence of ascorbic acid.

Conclusion: Ascorbic acid enhanced the development of porcine SCNT embryos via the increased TET3 mediated demethylation of somatic nucleus.

Citing Articles

- Invited Review - Reproductive technologies needed for the generation of precise gene-edited pigs in the pathways from laboratory to farm.

Tu C, Peng S, Chuang C, Wong C, Yang T Anim Biosci. 2022; 36(2):339-349.

PMID: 36397683 PMC: 9899582. DOI: 10.5713/ab.22.0389.

Utilization of Rosmarinic and Ascorbic Acids for Maturation Culture Media in Order to Increase Sow Oocyte Quality Prior to IVF.

Boldura O, Marc S, Otava G, Hutu I, Balta C, Tulcan C Molecules. 2021; 26(23).

PMID: 34885797 PMC: 8659116. DOI: 10.3390/molecules26237215.

Reactive oxygen and nitrogen species regulate porcine embryo development during pre-implantation period: A mini-review.

Luo Z, Yao J, Xu J Anim Nutr. 2021; 7(3):823-828.

PMID: 34466686 PMC: 8384778. DOI: 10.1016/j.aninu.2021.03.007.

References

Morgan H, Santos F, Green K, Dean W, Reik W . Epigenetic reprogramming in mammals. Hum Mol Genet. 2005; 14 Spec No 1:R47-58. DOI: 10.1093/hmg/ddi114. View

Wilmut I, Schnieke A, McWhir J, Kind A, Campbell K . Viable offspring derived from fetal and adult mammalian cells. Nature. 1997; 385(6619):810-3. DOI: 10.1038/385810a0. View

Portela A, Esteller M . Epigenetic modifications and human disease. Nat Biotechnol. 2010; 28(10):1057-68. DOI: 10.1038/nbt.1685. View

Tan L, Shi Y . Tet family proteins and 5-hydroxymethylcytosine in development and disease. Development. 2012; 139(11):1895-902. PMC: 3347683. DOI: 10.1242/dev.070771. View

Hemberger M, Dean W, Reik W . Epigenetic dynamics of stem cells and cell lineage commitment: digging Waddington's canal. Nat Rev Mol Cell Biol. 2009; 10(8):526-37. DOI: 10.1038/nrm2727. View

Huang Y, Tang X, Xie W, Zhou Y, Li D, Zhou Y . Vitamin C enhances in vitro and in vivo development of porcine somatic cell nuclear transfer embryos. Biochem Biophys Res Commun. 2011; 411(2):397-401. DOI: 10.1016/j.bbrc.2011.06.160. View

Zhao M, Kim N, Cui X . GlutaMAX prolongs the shelf life of the culture medium for porcine parthenotes. Theriogenology. 2015; 85(3):368-75. DOI: 10.1016/j.theriogenology.2015.08.014. View

Li E . Chromatin modification and epigenetic reprogramming in mammalian development. Nat Rev Genet. 2002; 3(9):662-73. DOI: 10.1038/nrg887. View

Kohli R, Zhang Y . TET enzymes, TDG and the dynamics of DNA demethylation. Nature. 2013; 502(7472):472-9. PMC: 4046508. DOI: 10.1038/nature12750. View

10.

Wakayama S, Mizutani E, Kishigami S, Van Thuan N, Ohta H, Hikichi T . Mice cloned by nuclear transfer from somatic and ntES cells derived from the same individuals. J Reprod Dev. 2005; 51(6):765-72. DOI: 10.1262/jrd.17061. View

11.

Niemann H, Tian X, King W, Lee R . Epigenetic reprogramming in embryonic and foetal development upon somatic cell nuclear transfer cloning. Reproduction. 2008; 135(2):151-63. DOI: 10.1530/REP-07-0397. View

12.

Jeong Y, Hossein M, Bhandari D, Kim Y, Kim J, Park S . Effects of insulin-transferrin-selenium in defined and porcine follicular fluid supplemented IVM media on porcine IVF and SCNT embryo production. Anim Reprod Sci. 2007; 106(1-2):13-24. DOI: 10.1016/j.anireprosci.2007.03.021. View

13.

Matoba S, Liu Y, Lu F, Iwabuchi K, Shen L, Inoue A . Embryonic development following somatic cell nuclear transfer impeded by persisting histone methylation. Cell. 2014; 159(4):884-95. PMC: 4243038. DOI: 10.1016/j.cell.2014.09.055. View

14.

Xu W, Wang Y, Li Y, Wang L, Xiong X, Su J . Valproic acid improves the in vitro development competence of bovine somatic cell nuclear transfer embryos. Cell Reprogram. 2012; 14(2):138-45. PMC: 3316478. DOI: 10.1089/cell.2011.0084. View

15.

Zhao M, Liang S, Guo J, Choi J, Kim N, Lu W . Analysis of Ferrous on Ten-Eleven Translocation Activity and Epigenetic Modifications of Early Mouse Embryos by Fluorescence Microscopy. Microsc Microanal. 2016; 22(2):342-8. DOI: 10.1017/S1431927616000040. View

16.

Yin R, Mao S, Zhao B, Chong Z, Yang Y, Zhao C . Ascorbic acid enhances Tet-mediated 5-methylcytosine oxidation and promotes DNA demethylation in mammals. J Am Chem Soc. 2013; 135(28):10396-403. DOI: 10.1021/ja4028346. View

17.

You J, Kim J, Lim J, Lee E . Anthocyanin stimulates in vitro development of cloned pig embryos by increasing the intracellular glutathione level and inhibiting reactive oxygen species. Theriogenology. 2010; 74(5):777-85. DOI: 10.1016/j.theriogenology.2010.04.002. View

18.

Lee K, Hamm J, Whitworth K, Spate L, Park K, Murphy C . Dynamics of TET family expression in porcine preimplantation embryos is related to zygotic genome activation and required for the maintenance of NANOG. Dev Biol. 2013; 386(1):86-95. DOI: 10.1016/j.ydbio.2013.11.024. View

19.

Walsh C, Bestor T . Cytosine methylation and mammalian development. Genes Dev. 1999; 13(1):26-34. PMC: 316374. DOI: 10.1101/gad.13.1.26. View

20.

Gu T, Guo F, Yang H, Wu H, Xu G, Liu W . The role of Tet3 DNA dioxygenase in epigenetic reprogramming by oocytes. Nature. 2011; 477(7366):606-10. DOI: 10.1038/nature10443. View