兔·卵·母·细·胞·孤·雌·激·活·的·研·究

Study on the Parthenogenetic Activation of Rabbit Oocytes

本研究探讨了兔卵母细胞孤雌激活的方法。兔卵母细胞经7%乙醇单独激活处理5min的卵裂率(13.0%),显著低于乙醇处理后在2mmol/LDMAP继续处理3h(48.1%)和5μmol/L离子霉素处理后在DMAP继续处理3h的卵母细胞(92.2%);离子霉素+DMAP处理组的囊胚发育率(27.1%)亦显著(P<0.05)高于乙醇+DMAP处理组(15.4%)和乙醇单独处理组(0%)。当用离子霉素和DMAP激活处理时,注射hCG后18h卵母细胞的卵裂率(92.1%)和囊胚率(35.3%)最高2,4h后卵母细胞的卵裂率(19.0%)和囊胚发育率(4.2%)均显著(P<0.05)下降。卵母细胞经离子霉素激活处理后,在DMAP继续处理1、35、h的卵裂率差异显著(P<0.05);而囊胚发育率无显著(P>0.05)差异。离子霉素和DMAP处理后再电激1次对卵母细胞的卵裂率(83.3%、80.0%)和囊胚率(30.0%、27.0%)无显著(P>0.05)影响。以上研究表明:离子霉素+DMAP是兔卵母细胞最有效的激活方法,注射hCG后18h的卵母细胞可取得较好的激活效果。

Parthenogene tic activation of rabbit oocytes was investigated in this study. The cleavage rate of oocytes activated in 7 % ethanol for 5 min （13.0%） were significantly lower than that of oocytes activated in 7 % ethanol （48.1%） or 5μmol/L ionomycin （92.2%） for 5 min following in 5 mmol/L DMAP for 3 h. The percentage of cleaved oocytes developing to blastocysts （27.1%） was also significantly higher （P 〈 0.05） when oocytes actvated in ionomycin and DMAP, in comparison with oocytes activated in ethanol alone （0%） or ethanol plus DMAP （15.4%）. When oocytes were activated using ionomycin and DMAP at 18 h after hCG injection, 92.1% of oocytes cleaved and 35.3% of cleaved oocytes developed to blastocysts, which were significantly higher than that of oocytes activated beyond 24 h post-hCG injection （19.0%, 4.2%, P 〈 0.05）. Three hours was proved to be appropriate for oocytes culturing in DMAP after ionomycin treatment, in which 88.6% of oocytes cleaved and 34.2% of cleaved oocytes developed to blastocysts, which was higher than that of oocytes culturing in DMAP for 1 h （86.1%, 28.3%） or 5 h （72.6%, 26.4%）. Additional electronic pulse to oocytes activated using ionomycin and DMAP did not affect their cleavage rate （83.3%, 80.0%） and blastocyst development （30.0%, 27.0%）. In conclusions, rabbit oocytes can be activated effectively by the treatment of ionomycin and DMAP, and the appropriate age of oocytes for parthenogenetic activation is at 18 h after HCG injection.