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Expression Analysis of Programmed Cell Death Patterns on the In-vitro Culture of Supplemented with Cell Death Regulation Factor during Development of Nuclear Transfer Embryos in Porcine

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한국동물생명공학회(구 한국수정란이식학회) (Journal of Animal Reproduction & Biotechnology)
초록

The nature of molecular mechanisms governing embryo development is largely unknown, but recent reports have demonstrated that proper execution of programmed cell death is crucial for this process. The main objective of this study is to examine the mode of programmed cell death during nuclear transfer embryos development in porcine. In particular, the relative employment of two major pathways in programmed cell death; e.g. apoptosis (type I) and autophagy (type II) was compared. Oocytes use in the study was matured in vitro in the presence of 10% FBS maturation medium. After nuclear transfer embryos were cultured for each programmed cell death control factor [Cysteamine(Cyst : 0.4mM), 3-methyladenine(3MA : 2.5mM) and Rapamycin(RP : 100nM)] in TCM-199 medium supplemented with 0.1% BSA. In this study results of among the blastocysts development in 3MA; PCNA, MAP1LC3A and ATG5 RNA gene expression level increased in the order of IVF<Cyst < 3MA < RP. However Casp-3 and TNF-r RNA gene expression level decreased in the order of IVF < 3MA and RP< Cyst. The expression of mTOR showed a pattern opposite to that of MAP1LC3A. That is, its expression was the lowest in Cyst group. And next experiments analysis of MMP expression patterns. Analysed this MMPs enzyme activation to evaluate the effectiveness of high quality brastocyst culture in porcine. In this results of the enzymatic activity of MMP-2 and MMP-9 was assessed in culture, the level of active MMP-9 was higher expression in the medium of each 3MA and RP treatment group, with the level of another treatment group being relatively higher. These results suggest that the autophagy activation culture medium is more effective for stable and innovative nuclear transfer embryos development.

저자
  • Na-Hyeon Jung(Department of Animal Life Science, Hankyong National University)
  • Sang-Hwan Kim(Institute of Genetic Engineering, Hankyong National University)
  • Enkhbolor(Major in the Animal Biotechnology The Graduate School of Biology & Information Technology, Hankyong National University)
  • Ji-Hye Lee(Major in the Animal Biotechnology The Graduate School of Biology & Information Technology, Hankyong National University)
  • Jong-Taek Yoon(Department of Animal Life Science, Hankyong National University Institute of Genetic Engineering, Hankyong National University)