不同交配时间对小鼠胚胎收集和发育的影响

Effect of different mating time on embryonic collection and embryonic development in mice

【目的】探索不同交配时间对小鼠胚胎收集和胚胎发育的影响,为小鼠的超排研究提供参考。【方法】对小鼠进行超数排卵,分别于注射人绒毛膜促性腺激素(hCG)后9,10,11,12,13,14和15h,在输卵管膨大部收集卵母细胞,研究排卵情况;将超排雌鼠与雄鼠合笼,分别于合笼后2,4,6,8,10和12h观察小鼠是否见阴道栓,研究小鼠交配时间;为了控制小鼠的交配时间,将超排雌鼠分别于注射hCG后0,2,4,6,8,10,12,14,16和18h与雄鼠合笼,每个时间点合笼后2h观察交配情况,于注射hCG后98h从子宫角收集囊胚,统计胚胎收集率,观察胚胎发育情况。【结果】小鼠排卵主要集中于注射hCG后12～14h,交配时间主要在注射hCG后的前6h。人为控制交配时间发现,注射hCG后4,6,8,10和12h的交配率均较高,较高的胚胎收集率出现在注射hCG后8,10和12h;囊胚发育率在注射hCG后4h(56.0%)和8h(54.5%)较高;扩张囊胚发育率在注射hCG后12h(68.2%)和16h(58.0%)显著高于其他时间点(P<0.05)。【结论】交配时间影响小鼠交配成功率、胚胎收集率和胚胎发育;超数排卵后12h交配可提高小鼠的胚胎收集率和胚胎发育率。

【Objective】 The effect of different mating time on mice embryonic collection and embryonic development by controlling mating time was investigated.【Method】 Ovulation was ensured by observing oocyte in oviduct at 9,10,11,12,13,14,15 h after injecting hCG.To observe the mating time,vaginal plug was recorded at 2,4,6,8,10,12 h after mating of superovulated female with male.To control the mating time,the female was caged with male mice at 0,2,4,6,8,10,12,14,16,18 h after injecting hCG.The mated mice were recorded at 2 h after caging,respectively.The embryo was collected in uterus at 98 h after injecting hCG.The collecting and development rate of embryo were recorded.【Result】 In nature,the ovulation occurred at 12-14 h after injecting hCG.The mating rate was mainly concentrated at first 6 h after caging.In control of mating time,mating rate was higher at 4,6,8,10,12 h after injecting hCG.The success of collecting embryo was higher at 8,10,12 h.Formation rate of blastocyst was higher at 4 h（56.0%） and 8 h（54.5%）;Development rate of expanded blastocyst was higher at 12 h（68.2%） and 16 h（58.0%）,respectively,which were significantly more than other points（P〈0.05）.【Conclusion】 The mating rate,collection and development of embryo was improved by controlling the mating time,and the rates of embryonic collection and development were improved by mating at 12 h after superovulating.