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수핵란의 전 활성화가 토끼 핵이식 수정란의 핵 재구성에 미치는 효과
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  • 수핵란의 전 활성화가 토끼 핵이식 수정란의 핵 재구성에 미치는 효과
  • Effect of Electrical Preactivation of Recipient Cytoplasm on Nuclear Remodelling in Nuclear Transplant Rabbit Embryos
저자명
전병균,김윤연,정기화,곽대오,이효종,최상용,박충생
간행물명
韓國家畜繁殖學會誌
권/호정보
1997년|21권 3호|pp.229-238 (10 pages)
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한국동물번식학회
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

Chromosome condensation and swelling of the donor nucleus have been known as the early morphological indicators of chromatin remodelling after injection of a foreign nucleus into an enucleated recipient cytoplasm. The effects of non-preactivation and electrical preactivation of recipient cytoplasm, prior to fusing a donor nucleus, on the profile of nuclear remodelling in the nuclear transplant rabbit embryos were evaluated. The embryos of 16-cell stage were collected and synchronized to G1 phase of 32-cell stage. The recipient cytoplasms were obtained by removing the first polar body and chromosome mass by non-disruptive microsurgical procedure. The separated G1 phase blastomeres of 32-cell stage were injected into non-preactivated recipient cytoplasms. Otherwise, the enucleated recipient cytoplasms were preactivated by electrical stimulation and the separated G1 phase blastomeres of 32-cell stage were injected. After culture until 20h post-hCG injection, the nuclear transplant oocytes were electrofused by electrical stimulation. The nuclei of nuclear transplant embryos fused into non-preactivated and/or preactivated recipient cytoplasm were stained by Hoechst 33342 at 0, 1.5, 2, 4, 6, 8, 10 hrs post-fusion and were observed under an fluorescence microscopy. Accurate measurements of nuclear diameter were revealed with an ocular micrometer at 200$ imes$. Upon blastomere fusion into non-preactivated recipient cytoplasm, a prematurely chromosome condensation at 1.5 hrs post-fusion and nuclear swelling at 8 hrs post-fusion were occurred as 91.6% and 86.1%, respectively. But the nuclei of nuclear transplant embryos fused into preactivated recipient cytoplasm, as o, pp.sed to non-preactivated recipient cytoplasm, were not occurred chromosome condensation and extensive nuclear swelling. Nuclear diameter fused into non-preactivated and preactivated recipient cytoplasm at hrs post-fusion was 30.2$pm$0.74 and 15.2$pm$1.32${mu}{ extrm}{m}$, respectively. These results indicated that onset of unclear condensation and swelling which was associated with oocytes activation were critical steps in the process of chromatin swelling. Futhermore, complete reprogramming seemed only possible after remodelling of the donor nucleus by chromosome condensation and nuclear swelling.