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目的 研究猪皮质颗粒在体外成熟及体外受精过程中的变化 ,以及皮质颗粒与多精受精的关系。 方法 用异硫氰荧光素联接的花生四烯酸 (FITC- PNA)标记体外成熟不同时间及体外受精不同时间皮质颗粒。 结果 皮质颗粒在猪卵母细胞体外成熟过程中逐渐由皮质部向卵质膜下迁移 ,并在质膜下排布成单层。体外成熟过程中所排出的第 1极体也被 FITC- PNA所标记 ,说明第 1极体中含有皮质颗粒。在对不同体外成熟时间的卵母细胞进行授精时发现 ,皮质颗粒的释放速度与体外成熟时间呈正比例关系。为进一步证实皮质颗粒的排放速度与多精受精之间的关系 ,还进行了精子穿透实验。结果显示 ,体外成熟 48h的卵母细胞其多精受精率为 73% ,比体外成熟36 h(83% )、2 4h(80 % )的卵母细胞要低。并且进入卵子的精子数目 (1.7)比后两者 (分别为 3.1和 2 .9)的少。 结论 多精受精率与皮质颗粒释放的速度和数量的大小有关
Objective To study the changes of porcine cortical granules during in vitro maturation and in vitro fertilization and the relationship between cortical granules and sperm fertilization. Methods FITC-PNA labeled with isothiocyanate-fluorescein was used to label cortical granules in vitro at different times of in vitro fertilization and in vitro fertilization. Results The cortical granules gradually migrated from the cortical part to the subepithelium during the in vitro maturation of porcine oocytes and arranged into a single layer under the plasma membrane. The first polar body discharged during in vitro maturation was also labeled with FITC-PNA, indicating that the first polar body contains cortical granules. In vitro fertilization of mature oocytes in vitro found that the release rate of cortical granules and in vitro maturation time was in direct proportion to the relationship. To further confirm the relationship between cortical particle release rate and fertilization, sperm penetration experiments were also performed. The results showed that the fertilization rate of oocytes matured in vitro for 48 h was 73%, which was lower than oocytes matured for 36 h (83%) and 24 h (80%) in vitro. And the number of sperm that entered the egg (1.7) was less than that of the latter two (3.1 and 2.9, respectively). Conclusion The fertilization rate is related to the speed and quantity of cortical granules released