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Asian-Australas J Anim Sci > Accepted
doi: https://doi.org/10.5713/ajas.16.0818    [Accepted]
Ascorbic Acid Increases Demethylation in Somatic Cell Nuclear Transfer Embryos of the pig (Sus scrofa)
Minghui Zhao , Tai-Young Hur, Jingu No, Yoonseok Nam, Hyeunkyu Kim, Gi-Sun Im, Seunghoon Lee*
National Institute of Animal Science, Jellabukdo, Korea
* Corresponding Author: Seunghoon Lee ,Tel: +82-632387272, Fax: +82-2387297, Email: sage@korea.kr
Received: October 21, 2016;  Revised: December 1, 2016.  Accepted: January 3, 2017.  Published online January 13, 2017.

ABSTRACT
Objective:
Investigated the effect and mechanism of ascorbic acid on the development of porcine embryos produced by somatic cell nuclear transfer (SCNT).
Method:
Porcine embryos were produced by SCNT and cultured in the presence or absence of ascorbic acid. TET3 in oocytes was knocked down by siRNA injection. After ascorbic acid treatment, reprogramming genes were analyzed by realtime RT-PCR. Furthermore, relative 5mC and 5hmC content in pronuclear 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, Pou5f1,Sox2 and Klf were significantly increased in blastocysts. Furthermore, ascorbic acid reduced 5-methylcytosine (5mC) content in pronuclar embryos compared with the control group. Knock down of TET3 in porcine oocytes significantly prevent the demethylation of somatic cell nuclear 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 nuclear.
Keywords: ascorbic acid; SCNT; TET3; demethylation, porcine
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