Reproductive and Developmental Biology

  • 제30권1호
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  • Pages.1-5
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  • 2006
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  • 1738-2432(pISSN)
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  • 2288-0151(eISSN)
한국동물번식학회 (The Korean Society of Animal Reproduction)
권호 표지

Chemical Agent를 이용한 추가 활성화 처리가 돼지 단위발생란의 발달에 미치는 영향

Development of Porcine Parthenogenetic Oocytes Activated with Different Combination of Chemicals

  • 서진성 (농촌진흥청 축산연구소, 순천대학교 동물자원과학과) ;
  • 황인선 (농촌진흥청 축산연구소) ;
  • 김세웅 (농촌진흥청 축산연구소) ;
  • 박효숙 (농촌진흥청 축산연구소) ;
  • 김동훈 (농촌진흥청 축산연구소) ;
  • 양병철 (농촌진흥청 축산연구소) ;
  • 공일근 (순천대학교 동물자원과학과) ;
  • 양보석 (농촌진흥청 축산연구소) ;
  • 임기순 (농촌진흥청 축산연구소)
  • 발행 : 2006.03.01
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초록

핵이식 방법을 이용하여 성공적인 복제를 이루기 위해서 인위적인 활성화 처리는 필수적인 요소이다. 본 연구는 전기자극에 의해 활성화된 난자를 chemical agent를 이용하여 추가적인 활성화 처리를 하였을 때 돼지 단위발생란의 발달에 미치는 영향을 알아보고자 수행되었다. 체외에서 $40{\sim}44$시간 동안 배양된 난자를 전기자극(E)으로 활성화 처리한 후 Thimerasol + Dithiothreitol(Thi+DTT), 6-Dimethylaminopurine(6-DMAP) 및 Cycloheximide(CH)를 사용하여 추가 활성화 처리를 하였다. 활성화 방법(E, E+Thi+DTT, E+6-DMAP 및 E+CH)에 따른 단위발생란의 배반포까지의 발달율을 조사한 결과, chemical agent에 의해 추가 활성화된 단위발생란이 전기자극만으로 처리된 구의 단위발생란보다 유의적으로 높은 발달율을 보였다($21.5{\sim}28.1%$ vs. 18.0%, P<0.05). 특히, E+Thi+DTT를 이용하였을 때 발달율이 유의적으로 높게 나타났다(28.1%, P<0.05). 활성화 처리별 전핵 형성율을 조사한 결과, chemical agent에 의해 추가 활성화 처리된 구에서 하나의 극체(1PN) 형성률은 처리별로 차이를 보이지 않았으나$(59.9{\sim}64.7%)$, 2PN 형성율은 추가 활성화 처리구에서 전기자극만을 사용하였을 때보다 유의적으로 높게 나타났다($7.2{\sim}9.7%$ vs. 4.3%, P<0.05). 이상의 결과를 살펴볼 때, 전기자극 후 chemical agent를 이용한 추가 활성화는 단위발생란의 배반포까지의 발달능력을 증가시키는 것으로 생각된다.

Artificial activation of oocytes is a prerequisite for the successful cloning by nuclear transfer. This study investigated the effect of the different combination of activation agents such as electric pulse (E), thimerosal (Thi) + dithiothreitol (DTT), 6-dimethylaminopurine (6-DMAP) or cycloheximide (CH) on the developmental ability of porcine embryos derived from parthenogenetic activation (PA). PA embryos activated with chemicals showed significantly higher developmental rate to the blastocyst stage compared to the embryos activated with E alone ($21.5{\sim}28.1%$ vs. 18.0%, respectively). Of chemicals, Thi + DTT supported higher development to the blastoryst stage (28.1%). There was no significant difference in 1 pronucleus (PN) formation rate $(59.9{\sim}64.7%)$, but 2PN formation rate was significantly higher in PA embryos with additional activation using chemicals $(7.2{\sim}9.7%)$. In conclusion, this study shows that chemical activation after electric pulse can increase the development of porcine PA embryos.

키워드

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