水牛精子在胞质内注射后的早期形态变化

Observation on the Morphological Changes of Buffalo Sperm Following ICSI

应用胞质内精子注射 (intracytoplasmic sperm injection,ICSI)技术探讨了水牛卵母细胞胞质与注射精子间的相互作用以及 ICSI精子的形态变化。ICSI后 13h,5 9.4 %的精子头部已膨大 ,且有 18.8%的精子进入解聚状态。雌雄原核发育不同步 ,雄原核在 ICSI后 16 h和 19h的形成率分别为 3.2 %和 4 0 .0 % ;雌原核在 ICSI后 13h已达到 71.9% ,16 h时提高到 90 .3%。经离子霉素与 6 - DMAP联合激活 ,ICSI后 19h产生的胚胎核型以 2 PN+PB1 为主 ,其雌原核形成 (激活 )率为 91.3% ,雄原核形成率为 4 0 .0 % ,5 1.3%为孤雌胚。用 5 mm ol/ L 的 DTT预处理精子 1h,可提高精子的解聚率 (30 .9%比 12 .7% ,P<0 .0 5 ) ,但对雄原核的形成率无显著影响 (33.3%比 32 .7% ,P>0 .0 5 )。结果表明 ,水牛精子 ICSI后的雄原核形成时间晚于卵子雌原核形成时间 ,且其比率低于卵子 ,DTT处理精子能提高其解聚率 。

The objective of this study was to examine the morphological changes of buffalo sperm interaction between oocytes cytoplasmic and intracytoplasmic sperm,morphological development of sperm following intracytoplasmic sperm injection(ICSI).At 13 h after ICSI,59.4% of the sperm head swelled and 18.8% of sperm shown partial decondensation.At 16 h and 19 h after ICSI,3.2% and 40.0% of oocytes had male pronucleus,while 71.9% and 90.3% of oocytes had formed female pronucleus at 13 h and 16 h after ICSI.At 19 h after activation treatment with ionomysin and 6-dimethylaminopurin,91.3% of ICSI oocytes activated(containing one or one more pronuclei),40.0% of ICSI oocytes shown normal fertilization(containing one male pronucleus and one female pronucleus),and 52.5% of them had double pronuclei.Pretreatment of spermatozoa with 5 μmol/L dithiothreitol(DTT) could promote the decondensation of sperm after ICSI(30.9% vs 12.7%,P<0.05),but did not affect the pronuclear formation(33.3% vs 32.7%,P>0.05).These results indicate that formation of female pronucleus is faster than that of male pronucleus,DTT can promote sperm decondensation but have no effect on the pronuclear formation.