Earticle

초록

영어

Cathepsin B is abundantly expressed peptidase of the papain family in the lysosomes, and closely related to the cell degradation system such as apoptosis, necrosis and autophagy. Abnormal degradation of organelles often occurs due to release of cathepsin B into the cytoplasm. Many studies have been reported that relationship between cathepsin B and intracellular mechanisms in various cell types, but porcine embryos has not yet been reported. Therefore, this study evaluated the effect of cathepsin B inhibitor (E-64) on preimplantation developmental competence and quality of porcine embryos focusing on apoptosis and oxidative stress. The expression of cathepsin B mRNA in porcine embryos was gradually decreased in inverse proportion to E-64 concentration by using real-time RT-PCR. When putative zygotes were cultured with E-64 for 24 h, the rates of early cleavage and blastocyst development were decreased by increasing E-64 concentration. However, the rate of blastocyst development in 5 μM treated group was similar to the control. On the other hand, both the index of apoptotic and reactive oxygen species (ROS) of blastocysts were significantly decreased in the 5 μM E-64 treated group compared with control. We also examined the mRNA expression levels of apoptosis related genes in the blastocysts derived from 5 μM E-64 treated and non-treated groups. Expression of the pro-apoptotic Bax gene was shown to be decreased in the E-64 treated blastocyst group, whereas expression of the anti-apoptotic Bcl-xL gene was increased. Taken together, these results suggest that proper inhibition of cathepsin B at early development stage embryos improves the quality of blastocysts, which may be related to not only the apoptosis reduction but also the oxidative stress reduction in porcine embryos.

목차

ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
  Chemicals
  In Vitro Maturation (IVM) and In Vitro Fertilization(IVF)
  In Vitro Culture (IVC)
  TUNEL Assay
  Measurement of ROS
  Gene Expression Analysis by Real-Time RT-PCR
  Statistical Analysis
 RESULTS
  Effect of E-64 on the Developmental Competence andCathepsin B Genes (CTSB) Expression of Porcine Blastocysts
  Apoptotic Patterns in Porcine Blastocysts Derived from E-64 Treated and Non-Treated Groups
  Effects of E-64 Treatment on Expression Levels of ROS in Porcine Blastocysts
 DISCUSSION
 REFERENCES

키워드

저자

• Hyeong-Hoon Son [ Department of Biotechnology, Daegu University, Gyeongsan 712-714, Korea ]
• Sung-Hun Min [ Department of Biotechnology, Daegu University, Gyeongsan 712-714, Korea ]
• Ji-Yeong Yeon [ Department of Biotechnology, Daegu University, Gyeongsan 712-714, Korea ]
• Jin-Woo Kim [ Department of Biotechnology, Daegu University, Gyeongsan 712-714, Korea ]
• Soo-Yong Park [ Department of Biotechnology, Daegu University, Gyeongsan 712-714, Korea ]
• Yong-Hee Lee [ Department of Biotechnology, Daegu University, Gyeongsan 712-714, Korea ]
• Pil-Soo Jeong [ Department of Biotechnology, Daegu University, Gyeongsan 712-714, Korea ]
• Deog-Bon Koo [ Department of Biotechnology, Daegu University, Gyeongsan 712-714, Korea ] Corresponding author

참고문헌

발행기관

간행물

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• 간행물명

  Reproductive & developmental biology

• 간기

  계간

• pISSN

  1738-2432

• eISSN

  2288-0151

• 수록기간

  1977~2018

• 십진분류

  KDC 527 DDC 636