Abstract
This experiment was carried out to investigate the optimal activation condition for parthenogenetic development. In order to activate oocytes at 22 h post onset of maturation, the oocytes were subjected to 5 $\mu$M ionomycin(I) for 5 min ,10 $\mu$M calcium ionophore(Ca) for 5 min, 2 mM 6-dimethylamino-purine(DMAP) for 3 h and 10 $\mu\textrm{g}$/$m\ell$ cycloheximide(CH) for 6 h alone or in combination. The activated oocytes were cultured in modified CR$_1$aa at 5% $CO_2$, 5% $O_2$, 90% $N_2$. l. The cleavage rates after 48 h culture of oocytes treated with I, Ca, DMAP and CH were 12.7%, 14.1%, 28.9% and 22.9%, respectively. There was no blastocyst formation. 2. The cleavage rates after 48 h culture of oocytes treated with I ＋ DMAP, I ＋ CH, Ca ＋ DMAP and Ca ＋ CH were 96.9%, 82.1%, 93.1% and 34.7%, respectively. Developmental rates to blastocysts were 10.4%, 5.3%, 17.6% and 7.1 %, respectively. When oocytes were treated with Ior Ca followed by DMAP, the blastocyst formation rate was significantly higher than other groups(P <0.05). 3. According to single activation treatment, pronucleus formation rates were 5.4%, 3.6%, 28.3% and 28.8%, respectively, Whereas, all oocytes treated with the combined activation agents formed 100% pronucleus.
활성화처리를 위해 22시간 성숙된 난자를 5 $\mu$M ionomycin(I)에서 5분, 10 $\mu$M calcium ionophore(Ca)에서 5분, 2 mM 6-dimethylaminopurine(DMAP)에서 3시간 및 10 $\mu\textrm{g}$/$m\ell$ cycloheximide(CH)에서 6시간 동안 단용 또는 병용 처리하였다. 활성화 처리된 난자는 mCR$_1$aa 배양액 내에서 배양되었으며, 배양조건은 5% $CO_2$, 95% air 또는 5% $CO_2$, 5% $O_2$, 90% $N_2$ 이었다. 1 I, Ca DMAP 및 CH에 의해 처리된 난자의 48시간 난할율은 각각, 12.7%, 14.1%, 28.9% 및 22.9%였다. 그러나, 배반포는 형성되지 않았다. 2. I＋DMAP, I＋CH, Ca＋DMAP및 Ca＋CH에 의해 처리된 난자의 48시간 난할율은 각각, 96.9%, 82.1%, 93.1% 및 84.7%였고, 배반포 발생율은 각각, 10.4%, 5.3%, 17.6% 및 7.1%로, I 및 Ca를 이용하여 세포 내 칼슘수준을 증가시킨 후, DMAP로 3시간 동안 배양하였을 때, 배반포 발생율이 유의적으로 높았다(P<0.05). 3. I, Ca, DMAP 및 CH에 의해 단용 처리된 난자의 전핵 발생율은 각각, 5.4%, 3.6%, 28.3% 및 28.8%였다. 그러나, 병용 처리된 난자는 100%의 전핵 형성율을 나타냈다.
