Earticle

초록

영어

This study was conducted to examine the optimal concentration and treatment time of antioxidants for inhibition of the ROS generation in bovine somatic cell nuclear transfer (SCNT) embryos. Bovine oocytes were activated parthenogenetically, during which oocytes were treated with various antioxidants to determine the optimal concentrations and kind of antioxidants. Determined antioxidants were applied to oocytes during in vitro maturation (IVM) and/or SCNT procedures. Finally, antioxidant-treated SCNT embryos were compared with in vitro fertilized (IVF) embryos. H2O2 levels were analyzed in embryos at 20 h of activation, fusion or insemination by staining of embryos in 10 μM 2'7'-dichlorodihydrofluorescein diacetate (H2DCFDA) dye, followed by fluorescence microscopy. H2O2 levels of parthenogenetic embryos were significantly lower in 25 μM β- mercaptoethanol (β-ME), 50 μM L-ascorbic acid (Vit. C), and 50 μM L-glutathione (GSH) treatment groups than each control group (24.0±1.5 vs 39.0±1.1, 29.7±1.0 vs 37.0±1.2, and 32.9±0.8 vs 36.3±0.8 pixels/embryo, p<0.05). There were no differences among above concentration of antioxidants in direct comparison (33.6±0.9～35.2±1.1 pixels/embryo). Thus, an antioxidant of 50 μM Vit. C was selected for SCNT. H2O2 levels of bovine SCNT embryos were significantly lower in embryos treated with Vit. C during only SCNT procedure (26.4±1.1 pixels/embryo, p<0.05) than the treatment group during IVM (29.9±1.1 pixels/embryo) and non-treated control (34.3±1.0 pixels/embryo). Moreover, H2O2 level of SCNT embryos treated with Vit. C during SCNT procedure was similar to that of IVF embryos. These results suggest that the antioxidant treatment during SCNT procedures can reduce the ROS generation level of SCNT bovine embryos.

목차

ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
  In Vitro Maturation of Oocytes
  Parthenogenesis
  Preparation of Donor Cells
  Nuclear Transfer
  Electrofusion and Activation
  Antioxidant Treatment
  In Vitro Fertilization
  Measurement of Intercellular ROS Content
  Statistical Analysis
 RESULS
  The H2O2 Levels of Parthenogenetic Embryos following Treatment with Various Concentrations of Antioxidants
  The H2O2 Levels of Parthenogenetic Embryos following Treatment with Different Antioxidants
  The H2O2 Levels of SCNT Embryos with Different Treatment Time of Antioxidant
  Comparison of H2O2 Levels between the SCNT Em bryosTreated with Antioxidant and IVF Embryos
 DISCUSSION
 REFERENCES

키워드

저자

• Hyo-Kyung Bae [ College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University ]
• Ji-Ye Kim [ College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University ]
• In-Sun Hwang [ College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University ]
• Choon-Keun Park [ College of Animal Life Sciences, Kangwon National University ]
• Boo-Keun Yang [ College of Animal Life Sciences, Kangwon National University ]
• Hee-Tae Cheong [ College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University ] 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