Study of Efficient Production of Cloned Embryos in Porcine II. Establishment of Conditional Enucleation

효율적인 돼지 복제수정란 생산에 관한 연구 II. 탈핵 여건의 확립

  • 위갑인 (전남대학교 농업생명과학대학 동물자원학부, 농업과학기술연구소) ;
  • 강만종 (전남대학교 농업생명과학대학 동물자원학부, 농업과학기술연구소) ;
  • 문승주 (전남대학교 동물자원학부)
  • Published : 2003.09.01

Abstract

This study was carried out to improve of enucleation efficiency on porcine recipient oocytes preactivated. In ethanol or $Ca^{2+}$ ionophore, effect of repeating and combinational activation with 6-DMAP or cycloheximide compared with alone activated treatment. Recipient oocytes's activation by $Ca^{2+}$ ionophore combined with 6-DMAP or cycloheximide were significantly higher than alone treatment(P<0.05). Between repeating and alone treatments were not significantly different. In ethanol, repeating treatment was significantly lower than alone(P<0.05), and combination treatments were not significantly different. On the basis of these results, efficiency of enucleation, electrical fusion and in vitro development compared preactivated with non-preactivated recipient oocytes. Enucleation and fusion rates of preactivated oocytes were improved significantly compared with non-preactivated oocytes(90.7%, 71.8 vs 77.8%, 61.1%; P<0.05). Behind the back, cleavage and in vitro development rates were significantly lower than non-preactivated oocytes(38.7%, 19.3% vs 68.8%, 30.6%; P<0.05).

체외 성숙시킨 M II기 난자의 여러 화학물질간의 중복 및 병용처리를 실시하여 수핵란으로서 적합한 활성화 조건과 활성화 후 자체 탈핵을 유도함으로써 보다 효율적인 탈핵 여건을 확립하고자 본 연구를 실시하였다. 1. Ethanol을 사용하여 6-DMAP 또는 cycloheximide 간의 단독, 중복, 그리고 병용처리를 실시한 결과, 활성화율, 난할율 및 체외 발달율에서 단독처리와 병용처리간의 차이는 없었으나 중복처리시 유의적으로 저하되었다(P<0.05). 2. $Ca^{2+}$ ionophore를 가지고 활성화 조건별로 처리하였을 경우, 6-DMAP와 병용 처리가 활성화율, 난할율 및 체외 발달율에서 각각 80.8%, 78.3%, 40.6%로 단독처리시의 58.0%, 62.9%, 27.0%보다 유의적으로 높았고(P<0.05), 또한 cycloheximide와의 병용처리도 활성화 및 난할율에서 단독처리와의 유의적 차이를 보였다(P<0.05). 그러나 중복처리시에는 효과를 나타내지 못하였다. 3. 탈핵전 수핵란에 미리 활성화 처리를 한 후 핵이식란을 재구성 한 결과 탈핵율 및 세포 융합율에서 90.7%, 71.8%로 활성화 처리하지 않은 것 77.8%, 61.1% 보다 유의적으로 높았다(P<0.05). 그러나 난할율 및 체외 발달율에서는 유의적으로 저하되었다(38.7%, 19.3% vs 68.8%, 30.6%, P<0.05). 또한 변성율도 활성화 처리하지 않은 구에 비해 유의적으로 높게 나타났다(P<0.05). 이상의 결과로 미루어 탈핵전 활성화하여 수핵란으로 이용할 경우 $Ca^{2+}$ ionophore와 6-DMAP를 병용 처리가 가장 효과적이며, 주입되는 체세포와 수핵란 간의 적정한 세포주기의 조절이 효율적인 돼지 복제수정란 생산을 위해 필요할 것이다.

Keywords

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