Earticle

초록

영어

This study investigated whether the addition of porcine sperm cytosolic factor (SCF) at fusion/activation affects in vitro development of porcine parthenogenetic(PA) and nuclear transfer (NT) embryos. To determine the optimum concentration of SCF, control group of oocytes was activated with 0.3M mannitol (1.0 mM CaCl2 ․ 2H2O), other three groups of oocytes were parthenogentically activated with the fusion medium (0.1mM CaCl2 ․ 2H2O) supplemented with 100, 200 or 300 μg/ml SCF, respectively. Matured oocytes were activated with two electric pulses (DC) of 1.2 kv/cm for 30 μsec. The activated embryos were cultured in PZM-3 under 5% CO2 in air at 38.5℃ for 6 days. Oocytes activated in the presence of SCF showed a significantly higher blastocyst rate than control (p<0.05). Apoptosis rate was significantly lower in 100 μg/ml SCF group than other groups (p<0.05). Cdc2 kinase activity in control and SCF treatment group of oocytes was determined using MESACUP cdc2 kinase assay kit at 1, 5, 10, 15, 30, 45 and 60 min after activation. Cdc2 kinase activity was significantly decreased (p<0.05) in SCF group than MII oocytes or control within 5 min. For NT embryo production, reconstructed oocytes were fused in the fusion medium supplemented with 0.1 mM CaCl2 ․ 2H2O (T1), 1.0 mM CaCl2 ․ 2H2O (T2) and 0.1 mM CaCl2 ․ 2H2O with 100 μg/ml SCF (T3). Fused embryos were cultured in PZM-3 under 5% CO2 in air at 38.5℃ for 6 days. Developmental rate to blastocyst stage was significantly higher in T3 than other groups (23.0% vs. 13.5 to 15.2%) (p<0.05). Apoptosis rate was significantly lower in T3 than T1 or T2 (p<0.05). The relative abundance of Bax-α/Bcl-xl was significantly lower in in vivo or SCF group than that of control (p<0.05). Moreover, the expression of p53 and caspase3 mRNA was significantly lower in in vivo or SCF group than that of control (p<0.05). These results indicate that the addition of SCF at fusion/activation might improve in vitro development of porcine NT embryos through regulating cdc2 kinase level and expression of apoptosis related genes.

목차

ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
  Preparation of SCF
  Collection of Oocytes and In Vitro Mmaturation (IVM)
  Production of Parthenogenetic (PA) Embryos
  Preparation of Porcine Fetal Fibroblast Cells
  Production of Nuclear Transfer (NT) Embryos
  Apoptosis Assays
  Collection of In Vivo Blastocysts
  RealTime RT-PCR Quantification
  Cdc2 Kinase Assay
  Experimental Designs
  Statistical Analysis
 RESULTS
  Development of PA Embryos Produced in the Presence of SCF
  Cdc2 Kinase Activity
  Development of NT Embryos Produced in the Presence of SCF
  Expression of Apoptosis-Related Genes in NT Embryos
 DISSCUSSION
 REFERENCES

키워드

저자

• Joo-Hyun Shim [ Animal Biotechnology Division, National Institute of Animal Science, RDA, Chungnam National University ]
• Dong-Hoon Kim [ Animal Biotechnology Division, National Institute of Animal Science, RDA ]
• Yeoung-Gyu Ko [ Animal Biotechnology Division, National Institute of Animal Science, RDA ]
• Seong-Soo Hwang [ Animal Biotechnology Division, National Institute of Animal Science, RDA ]
• Keon-Bong Oh [ Animal Biotechnology Division, National Institute of Animal Science, RDA ]
• Boh-Suk Yang [ Animal Biotechnology Division, National Institute of Animal Science, RDA ]
• Dong-Il Jin [ Chungnam National University ]
• Jin-Ki Park [ Animal Biotechnology Division, National Institute of Animal Science, RDA ]
• Gi-Sun Im [ Animal Biotechnology Division, National Institute of Animal Science, RDA ] Corresponding author

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  Reproductive & developmental biology

• 간기

  계간

• pISSN

  1738-2432

• eISSN

  2288-0151

• 수록기간

  1977~2018

• 십진분류

  KDC 527 DDC 636