细胞骨架抑制剂、电激活液对兔卵孤雌激活和发育的影响

The Effects of Cytoskeleton Inhibitor and Electro-activation Solution on the Parthenogenetic Activation and Preimplantation Development in Rabbit Oocytes

应用细胞骨架抑制剂细胞松弛素B和秋水仙素处理兔成熟卵母细胞,然后采用电激活的方法激活兔卵母细胞,观察细胞骨架抑制剂对兔卵母细胞孤雌激活和孤雌发育的影响,结果表明:应用不同的电激活液,即甘露醇、Zimmerman氏和山梨醇对兔卵母细胞的孤雌激活效果无显著性差异(P>0.05);用7.5μg.mL-1细胞松弛素B处理卵母细胞后孤雌激活,其2-细胞胚率(82.2%)和囊胚发育率(43.4%)与对照组(76.4%和51.5%)相比无显著性差异(P>0.05);用1μg.mL-1秋水仙素处理兔卵母细胞,兔卵母细胞孤雌激活后的2-细胞胚率(64.9%)和囊胚发育率(23.8%)明显下降,与对照组相比存在显著性差异(P<0.05);用细胞松弛素B和秋水仙素共同处理的2-细胞胚率(62.5%)和囊胚发育率(30.9%)与秋水仙素单独处理的结果无显著性差异(P>0.05)。因此,秋水仙素对兔卵母细胞的孤雌激活影响比CB更大。通过免疫荧光和激光扫描共聚焦显微镜观察发现,细胞骨架抑制剂处理后,经孤雌激活获得的囊胚中,微丝和微管结构未见异常。

The present study aimed to investigate effects of cytoskeleton inhibitors and electro-activation solutions on the parthenogenetic activation and preimplantation development in rabbit. MII oocytes were treated with cytochalasin B or cochicines before electro-activation. The results showed that there was no significant different among electro-activation solutions considering oocytes parthenogenetic activation （P〉0.05）. Furthermore there was no significant difference of 2- cell or blastocyst development when rabbit oocytes were treated with or without cytochalasin B （7.5 μg · mL^-1） before electro-activation （82.2% vs 76.4%, and 43.4% vs 51.5%, respectively）. （P〉0.05）. The developmental rate of 2-cell and blastocyst were significantly decreased （64.9% and 23.8% respectively） when oocytes were treated with colchicines（1.0 μg · mL^-1） before electro-activation comparing with that of no treatment （P-0.05）. The results from cytocha- lasin B plus cochicine treatment were similar to that of cochicines treated alone （62.5% 2-cell rate and 30.9% blastocyst rate）. Immunofluorescent staining and laser scanning confocal microscopy results indicated that there was no abnormality of microtubule or microfilament in blastocysts which derived from cytoskeleton inhibitor treated and following electronic activated oocytes.