CAJ | 학술논문

目的：探寻一种稳定有效的卵母细胞激活方法，为兔体细胞核移植提供实验基础。方法：兔超排后获得卵母细胞体外成熟22～24 h，经脱颗粒处理后采用不同激活剂和不同处理时间进行激活，体外培养5～7 d，观察胚胎发育与囊胚形成情况。结果：卵母细胞经过离子霉素作用5 min或者乙醇作用7 min均能被激活，但两者卵裂率、囊胚率都较低；乙醇和离子霉素分别与6-DMAP联合激活兔卵母细胞时，离子霉素＋6-DMAP 处理组卵裂率（73．7％）显著高于乙醇＋6-DMAP处理组（38．5％，P＜0．05）；与离子霉素联合激活时，2．0 mmol/L的6-DMAP处理5．0 h时的卵裂率和囊胚率最高，达75．6％和45．5％。结论：激活方法对兔卵母细胞孤雌激活率的影响很大，不同激活剂的联合应用及激活剂的作用时间对兔卵母细胞的激活存在显著差异。
목적：탐심일충은정유효적란모세포격활방법，위토체세포핵이식제공실험기출。방법：토초배후획득란모세포체외성숙22～24 h，경탈과립처리후채용불동격활제화불동처리시간진행격활，체외배양5～7 d，관찰배태발육여낭배형성정황。결과：란모세포경과리자매소작용5 min혹자을순작용7 min균능피격활，단량자란렬솔、낭배솔도교저；을순화리자매소분별여6-DMAP연합격활토란모세포시，리자매소＋6-DMAP 처리조란렬솔（73．7％）현저고우을순＋6-DMAP처리조（38．5％，P＜0．05）；여리자매소연합격활시，2．0 mmol/L적6-DMAP처리5．0 h시적란렬솔화낭배솔최고，체75．6％화45．5％。결론：격활방법대토란모세포고자격활솔적영향흔대，불동격활제적연합응용급격활제적작용시간대토란모세포적격활존재현저차이。
The objective of this study is to seek a stable and effective method for the parthenogenetic activation of rabbit oo -cytes, and make an experimental foundation for the somatic cell nuclear transfer of rabbit .Cumulus-oocyte complexes were harves-ted from the ovaries of superovulated rabbits , and were matured in vitro for 22~24 h;after removing the cumulus cells , the oocytes were activated by various stimuli for different time .After in vitro culture for 5~7 d, the parthenogenetic development of stimulated oocytes was assessed by microscopic observation .The results showed that rabbit oocytes could be activated not only by ionomycin treatment for 5 min, but also by ethanol treatment for 7 min, but their cleavage rate and blastula rate all were relatively low .When 6-DMAP combined with ionomycin or ethanol was used to activate rabbit oocytes , the cleavage rate (73.7%) in the treatment of ionomycin +6-DMAP was significantly (P<0.05) higher than that (38.5%) in the treatment of ethanol +6-DMAP; the treatment of ionomycin +2.0 mmol/L 6-DMAP for 5.0 h obtained the highest cleavage rate and blastula rate , being 75.6%and 45.5%respectively.It was concluded that activation method had great influence on the parthenogenetic activation rate of rabbit oo -cytes, and there were significant differences in activation effect among different treatments (combined utilization of different stimuli , various activation time ) .