Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
권호 표지

$In$ $Vitro$ Development of Goat Parthenogenetic Oocytes Derived from Different Activation Methods

도축장 유래 산양난자의 단위 발생 유기 방법에 따른 체외 발달

  • Yun, Yun-Jin (Department of Animal Science and Biotechnology, Gyeongnam National University of Science and Technology) ;
  • Park, Kyeong-Jin (Department of Animal Science and Biotechnology, Gyeongnam National University of Science and Technology) ;
  • Park, Hee-Sung (Department of Animal Science and Biotechnology, Gyeongnam National University of Science and Technology)
  • 윤윤진 (경남과학기술대학교 동물생명과학과) ;
  • 박경진 (경남과학기술대학교 동물생명과학과) ;
  • 박희성 (경남과학기술대학교 동물생명과학과)
  • Received : 2012.01.10
  • Accepted : 2012.01.24
  • Published : 2012.03.31
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Abstract

Efficient oocyte activation is a key step for the success of nuclear transfer in cloning. Ionomycin sequentially combined with 6-DMAP is now widely used to activate normal oocytes for analytical studies of oocyte activation and to activate reconstructed oocytes after nuclear transfer. The present study investigated sources of oocytes, duration of ionomycin and 6-DMAP, laser and electric stimulation in goat oocyte activation in order to optimize the protocols. Goat ovaries were collected in individual abattoirs during the breeding season and were delivered to the laboratory within 6 h in saline with 100 IU/ml streptomycin and 0.05 mg/ml penicillin. The oocytes were denuded from the cumulus cell by pipetting with 0.2% hyaluronidase in PBS at 20~22 hr post maturation. Oocytes with the polar body were selected and assigned to four groups for parthenogenetic activation. To examine the effect of duration of ionomycin treatment, oocytes after 20~22 hr of maturation were treated with 2.5 uM ionomycin for 1 or 5 min times and then cultured in 2 mM 6-DMAP for 2 or 4 hr. The activated oocytes were cultured in mSOF at $38.5^{\circ}C$ in $CO_2$ 5%, $O_2$ 5% and $N_2$ 90% multi incubator. Cleavage and blastocyst development was observed at 48 hr and day 8 of culture $in$ $vitro$, respectively. Activation rates of oocytes exposed to ionomycin for 1 min(86.4%) were significantly higher than those treated for 5 min(74.3%) duration. This indicated that 1 min ionomycin treatment was most suitable for activation of goat oocytes. The duration of 6-DMAP treat duration was in 2 mM 6-DMAP for 2 hr after 1 min exposure to 2.5 uM ionomycin. The activation rate of oocytes incubated in 6-DMAP for 2 hour(82.5%) was significantly higher than those in oocytes treated with 4 hr(75.5%).

Keywords

References

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