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牛卵胞质内显微受精中第一极体位置与激活方法研究
原巨强1,2, 陈静波2, 赵云程2,3
1.西北农林科技大学 动物医学院 农业部家畜生殖内分泌与胚胎工程重点开放实验室;2.新疆畜牧科学院;3.新疆农业大学 动物科技学院
摘要:
[目的]探讨牛卵母细胞胞质内显微受精(ICSI)操作中,卵母细胞第一极体位置以及受精卵化学激活方法对ICSI卵体外发育的影响.[方法]采集牛卵巢,成熟培养卵母细胞后,调整其第一极体分别位于相当于时钟6点和12点的位置,进行牛卵母细胞胞质内显微受精,以ICSI卵不进行激活处理为未激活组,分别采用乙醇、离子霉素及离子霉素联合6-二甲基氨基嘌呤(6-DMAP)激活ICSI卵,统计ICSI卵的形态完整率、卵裂率和囊胚发育率.[结果]第一极体分别位于相当于时钟6点和12点的位置时,所获得的ICSI卵形态完整率(92.9%和94.0%)、卵裂率(37.1%和40.0%)和囊胚发育率(12.9%和12.2%)均无统计学差异(P>0.05).未激活组ICSI卵的卵裂率较低,不能发育到囊胚;用乙醇或离子霉素激活时,ICSI卵的卵裂率(25.7%和27.6%)和囊胚发育率(3.3%和3.4%)均显著高于未激活组 (7.5%和0)(P<0.05);用离子霉素与6-DMAP联合激活时,ICSI卵的卵裂率和囊胚发育率分别为38.9%和13.9%,显著高于乙醇激活组、离子霉素激活组及未激活组(P<0.05).[结论]对牛卵母细胞进行ICSI操作时,第一极体分别位于相当于时钟6点和12点位置时,对ICSI卵的发育无显著影响;采用离子霉素和6-DMAP联合激活,有利于ICSI卵的体外发育.
关键词:  牛卵胞质内显微受精  第一极体  化学激活  囊胚发育率
DOI:
分类号:
基金项目:陕西省重大科技专项计划项目(2006KZ07-G1);西北农林科技大学科研专项(04ZM001)
Studies on direction of the first polar body and different activation methods in bovine oocytes intracytoplasmic sperm injection
Abstract:
【Objective】 This study investigated the effect of direction of the first polar body (PBⅠ) and different activation method on the development of the bovine intracytoplasmic sperm injection (ICSI) zygote in vitro.【Method】 Oocytes were collected from ovaries and matured in vitro,two directions ( 6 and 12 o'clock ) of the first polar body modulated in oocytes at ICSI were examined,then oocytes after ICSI were treated with ethanol,Ionomycin and Ionomycin combine with 6-Dimethylaminopurine (6-DMAP) for activation,or without activating treat (the control group),the ICSI zygotes modality integrity rate,cleavage rate and blastocyst rate were counted. 【Result】 There were no significant differences in the ICSI zygotes modality integrity rate (92.9% vs.94.0%,P>0.05),cleavage rate (37.1% vs. 40.0%,P>0. 05),blastocyst rate (12.9% vs.12.2%,P>0. 05) between the two directions of the PBⅠ.The ICSI zygotes cleavage rate and blastocyst rate were higher with activated by 7% ethanol (25.7%,3.3%) and Ionomycin(27.6%,3.4%) than that with the control group (7.5%,0) (P<0.05),the ICSI zygotes cleavage rate and blastocyst rate were significantly higher with activated by Ionomycin combine with 6-DMAP (38.9%,13.9%) than those with other groups (P<0.05).【Conclusion】 In conclusion,there were no significant differences in the ICSI zygotes modality integrity rate,cleavage rate,blastocyst rate between the two directions ( 6 o'clock and 12 o'clock ) of the the PBⅠ,activated with Ionomycin followed by 6-DMAP could improve the development of ICSI zygotes.
Key words:  bovine intracytoplasmic sperm injection  first polar body  chemical activation  blastocyst rate