Somatic cell nuclear transfer (SCNT) is a useful biotechnological tool for animal cloning. Until now, SCNT has been inefficient, especially in dog. It is believed that an embryo developmental block in SCNT embryos is cause of low production efficiency. However, no studies have been performed on canines for embryo developmental block. In this study, we attempted to evaluate the beneficial role of EDTA in canine parthenogenic (PA) embryos development to overcome embryo developmental block. The PA embryos were divided into 0.01 mM EDTA treated and non-treated groups. Embryo developmental efficiency was measured by activating chemically parthenote. After EDTA induction, PA embryos were evaluated for embryonic development, Reactive Oxygen Species (ROS) activity, mitochondrial integrity, ATP production and genomic activation. The EDTA treated PA embryos showed significantly higher survival rate and improved cavity formation compared to non-treated. Furthermore, cytoplasmic ROS level was mitigated and mitochondrial membrane potential was found significantly higher in EDTA treated group followed by higher ATP production. Moreover, major embryonic genomic activation specific markers/factors were also elevated in EDTA treated group. Conclusively, we elucidated that EDTA showed substantially positive effect to overcome embryo developmental block in canine.

Abstract
 서 론
 재료 및 방법
  1. 시약 및 배양액
  2. 체내성숙 된 개 난자의 채취와 Pathenogenesis(PA)
  3. 개 PA 배아의 세포 수 측정
  4. 개 PA 배아에서 Cytoplasmic Reactive Oxygen Species (ROS) level 의 측정
  5. JC-1 staining
  6. ATP concentration detection
  7. 핵 관련 유전자 정량분석 (Real-time PCR)
  8. 통계 처리
 결과
  1. 세포수, 생존율 및 cavity formation에 대한 EDTA의 효과
  2. EDTA 처리에 따른 cytoplasmic ROS level
  3. mitochondrial integrity를 확인하기 위한 JC-1 staining
  4. ATP production
  5. Genomic activation
 고찰
 REFERENCES

• Haeyun Jeong(Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration&2Animal Reproduction Laboratory, College of Agriculture and Life Science, Chonbuk National University) | 정해윤
• Minghui Zhao(Qingdao Agricultural University 700 Changcheng Rd, Chengyang Qu, Qingdao Shi, Shandong Sheng, China) | Zhao Minghui
• Jin-Gu No(Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) | 노진구
• Imran Ullah(Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) | Ullah Imran
• Whi-Cheul Lee(Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) | 이휘철
• Hayeon Wi(Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) | 위하연
• Sun A Ock(Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) | 옥선아
• Jae-Seok Woo(Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) | 우제석
• Tai-young Hur(Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) | 허태영
• Gi-sun Im(Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) | 임기순
• Jong-Gug Kim(Animal Reproduction Laboratory, College of Agriculture and Life Science, Chonbuk National University) | 김종국
• Seunghoon Lee(Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) | 이승훈 Corresponding Author