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Effect of Osmolarity of Culture Medium on Imprinting and Apoptotic Gene Expression in Miniature Pig Nuclear Transfer Embryos

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  • URLhttps://db.koreascholar.com/Article/Detail/123589
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한국동물번식학회 (The Korean Society of Animal Reproduction)
초록

This study was conducted to investigate the development and gene expression in miniature pig nuclear transfer (mNT) embryos produced under different osmolarity culture conditions. Control group of mNT embryos was cultured in PZM-3 for 6 days. Treatment group of mNT embryos was cultured in modified PZM-3 with NaCl (mPZM-3, 320 mOsmol) for 2 days, and then cultured in PZM-3 (270 mOsmol) for 4 days. Blastocyst formation rate of the treatment group was significantly higher than the control and the apoptosis rate was significantly lower in treatment group. Bax- and caspase-3 mRNA expression were significantly higher in the control than the treatment group. Also, the majority of imprinting genes were expressed aberrantly in in vitro produced mNT blastocysts compared to in vivo derived blastocyst H19 and Xist mRNA expression were significantly lower in the control than the treatment group or in vivo. IGF2 mRNA expression was significantly higher in the control than the treatment group or in vivo. IGF2r mRNA expression was significantly lower in the control. Methylation profiles of individual DNA strands in H19 upstream T-DMR sequences showed a similar methylation status between treatment group and in vivo. These results indicate that the modification of osmolarity in culture medium at early culture stage could provide more beneficial culture environments for mNT embryos.

목차
ABSTRACT   INTRODUCTION   MATERIALS AND METHODS    Oocyte Recovery and Maturation    Preparation of Nuclear Donor Cells    Production of Nuclear Transfer Embryos    Terminal DeoxynucleotidyI Transferase-Mediated dUTP Nick-End Labeling (TUNEL) Assay    Collection of In Vivo Blastocysts    Realtime PCR Quantification    Statistical Analysis   RESULTS   DISCUSSION   REFERENCES
저자
  • Mi-Rung Park(Division of Animal Biotechnology, National Institute of Animal Science, RDA)
  • In-Sun Hwang(Division of Animal Biotechnology, National Institute of Animal Science, RDA)
  • Joo-Hyun Shim(Division of Animal Biotechnology, National Institute of Animal Science, RDA)
  • Hyo-Jin Moon(Division of Animal Biotechnology, National Institute of Animal Science, RDA)
  • Dong-Hoon Kim(Division of Animal Biotechnology, National Institute of Animal Science, RDA)
  • Yeoung-Kyu Ko(Division of Animal Biotechnology, National Institute of Animal Science, RDA)
  • Hwan-Hoo Seong(Division of Animal Biotechnology, National Institute of Animal Science, RDA)
  • Gi-Sun Im(Division of Animal Biotechnology, National Institute of Animal Science, RDA) Corresponding author