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핵이식 기법을 이용한 한우 쌍태생산에 관한 연구 I. Ovum pick-up(OPU), 전기적 세포융합 및 체외배양 기법을 이용한 복제수정란 생산
황우석,신태영,노상호,이병천,Hwang, Woo-suk,Shin, Tae-young,Roh, Sang-ho,Lee, Byeong-chun 대한수의학회 1998 大韓獸醫學會誌 Vol.38 No.1
The purpose of present study is to improve the efficiency of fusion and the developmental rates of nuclear transplanted embryos to produce genetically identical twins from Korean native cattle. The diameter of aspirated follicles had no significant effect on the recovery rates of oocytes collected by ovum pick-up technique. The fusion rates of nuclear transplanted embryos were significantly higher in 50 and $100{\mu}s$ DC duration groups(73.3 and 72.0% ; respectively) than that in $30{\mu}s$ group(55.6% ; p<0.05). The cleavage rates of nuclear transplanted embryos appeared to be significantly higher in donor nuclei derived from in vivo (65.0%) than in those from in vitro (50.5% ; p<0.01), but the developmental rates to morulae and blastocysts were not significantly different between them(13.7 vs 10.9%, respectively).
핵이식 기법을 이용한 한우 쌍태생산에 관한 연구 II. Ovum pick-uo(OPU) 유래 공여핵 및 활성화 유도 수핵난자의 핵이식
황우석,신태영,노상호,박종임,이병천,Hwang, Woo-suk,Shin, Tae-young,Roh, Sang-ho,Park, Jong-im,Lee, Byeong-chun 대한수의학회 1998 大韓獸醫學會誌 Vol.38 No.3
The efficiency of nuclear transfer using donor embryos originated from ovum pickup(OPU) and activated recipient cytoplasts were examined for induction of twinning in Korean native cattle(KNC). After aspiration of follicle by OPU, regardless of the vacuum applied, we obtained same result in proportions of recovered cumulus-oocyte complex (COCs) with compact cumulus. Under electric stimulation(1.0kV/cm DC for $40{\mu}s$), most of activated oocytes proceed to anaphase II/telophase II within 3h(84.7%). In the treatment of oocyte activation, the preactivation which was performed before fusion had significant effect on the developmental rates to morula/blastocyst stage(9.4 vs 4.0%). In embryo transfer of nuclear transferred embryos, we obtained 2 twins from KNC recipients and 1 twin from a Holstein recipient. Our results showed that it is possible to obtain twins using nuclear transfer technique in KNC.
컴퓨터 정액자동분석에 의한 동결융해 한우 정액의 운동특성 연구
이강남,이병천,김정태,박종임,신태영,황우석,Lee, Kang-nam,Lee, Byeong-chun,Kim, Jung-tae,Park, Jong-im,Shin, Tae-young,Hwang, Woo-suk 대한수의학회 1998 大韓獸醫學會誌 Vol.38 No.4
The aim of this experiments were to assess the time-interval change of motional characteristics in frozen-thawed semen of Korean native cattle (KNC) by using computer aided semen analysis (CASA) technology. Twenty-six KNC frozen semen straws were obtained from Korean KNC improvement department, livestock improvement main division, national livestock cooperatives federation in Korea. Specimens were allowed to thaw at $37^{\circ}C$ for 30 sec in water bath. Semen analysis was performed on semen image analysis system (SIAS, Medical supply, Korea) adjusted to the gate settings and used the semen droplet ($5{\mu}l$) placed on Makler counting chamber (Sefi medical instrument, Israel) prewarmed at $37^{\circ}C$. The same person used the same micropipette to fill the Makler counting chamber. A total of 150 or more of sperms were analysed in each specimen by a single trained person by scanning at least 5 to 10 fields. The measurement parameters in SIAS were as follows ; frame rate = 30 frames per sec, image capture = 1 sec, minimum motile speed = $10{\mu}m/s$, maximum countable sperm number = 400. Statistical analysis was done by Student t-test with use of the Sigma plot program on a IBM personal computer. The dancemean(DNM) and hyperactivated sperm(HYP) of frozen-thawed KNC semen kinematics were significantly decreased(p < 0.05) after 10 min of incubation at $37^{\circ}C$ water bath. But, wobble(WOB) of same sample semen was significantly increased(p < 0.05) after 10 min of incubation and significantly decrease(p < 0.05) after 60 min of same incubation. And, after 30 mim of incubation, significantly differences were found most of motion kinematics, motifity(MOT), curvilinear velocity(VCL), straight line velocity(VSL), average path velocity(VAP), amplitude of lateral head displacement(ALH), beat cross frequency(BCF), mean angular displacement(MAD), dance(DNC), on same sample semen. The DNM of KNC semen sample was variable kinematics after 30 min of incubation. Also, the linearity(LIN) and straightness(STR) was significantly decreased(p < 0.05) from 60 min of incubation. In conclusion, the AI within 30 min after thawing of frozen semen can be an effective method for obtaining high fertility rate in KNC reproductive program.