人工激活胚胎的性染色体分析和IGF-Ⅱ表达研究  

Sex chromosome analysis and IGF-Ⅱ expression on activated human unfertilized oocytes after ICSI with calcium ionophore A23187 and puromycin

作　　者：鹿群[1] 陈子江[1] 高选[1] 李媛[1] 颜军昊[1] 马水英[1] 李梅[1] 

机构地区：[1]山东大学山东省立医院生殖医学中心

出　　处：《中华医学遗传学杂志》2005年第5期524-527,共4页Chinese Journal of Medical Genetics

基　　金：国家自然科学基金(39300136和30270512)~~

摘　　要：目的观察钙离子载体A23187联合嘌呤霉素激活胚胎的性染色体和胰岛素样生长因子-Ⅱ(insulin-like growthfactor-Ⅱ,IGF-Ⅱ)表达。方法收集体外成熟周期(in vitromaturation and intracytoplasmicsperminjection,IVM-ICSI)和卵母细胞单精子显微注射(intracytoplasmic sperminjection,ICSI)中受精失败的卵母细胞95枚,采用钙离子载体A23187和嘌呤霉素激活。应用荧光原位杂交技术分析来源于2PN2PB胚胎的性染色体;免疫组化检测IGF-Ⅱ的表达,并与正常胚胎、孤雌胚胎相比较。结果钙离子载体A23187联合嘌呤霉素能有效地激活ICSI后22h未受精卵母细胞,激活胚胎能发育到囊胚阶段。激活胚胎的性染色体为5枚XX,8枚XY。激活胚胎的IGF-Ⅱ表达与正常胚胎相近,明显较孤雌胚胎增强。结论钙离子载体A23187联合嘌呤霉素是一种安全、有效的激活方式,有希望成为ICSI受精失败的有效补救措施。Objective To investigate the sex chromosomes and the expression of insulin-like growth factor-Ⅱ （IGF-Ⅱ ） on activated human unfertilized oocytes after intracytoplasmic sperm injection（ICSI） with calcium ionophore A23187 and puromycin. Methods All 95 discarded oocytes that showed no evidence of fertihzation at 16-18 h after in vitro maturation and intracytoplasmic sperm injection cycles （IVM-ICSI）/conventional ICSI were exposed to calcium ionophore A23187 （5μmot/L） for 5 min and then were incubated with puromycin （ 10μg/mL） for 4 h. After activation, the oocytes were cultured in vitro for 3-5 days. The sex chromosome analysis was performed by dual color fluorescence in situ hybridization. The expression of IGF-Ⅱ on the activated embryos, normal embryos, and parthenotes was examined. Results The combination of calcium ionophore A23187 with puromycin could activate the unfertilized oocytes 22 h after ICSI. The activated rate, cleavage rate, and quality of activated embryos of the IVM-ICSI group were similar to those of ICSI group, respectively. Sex chromosome analysis indicated that 8 male and 5 female embryos had been derived from two pronucleus and a second polar body. The expression of IGF-Ⅱ on activated embryos and normal embryos was high and similar, which was much stronger than that of parthenotes. Conclusion The combination of calcium ionophore A23187 with puromycin could effectively activate unfertilized oocytes 22 h after ICSI. Moreover, the unfertilized oocytes activated by calcium ionophore A23187 and puromycin had normal sex chnromosomes and expression of IGF-Ⅱ like the normal embryos. These suggest that oocyte activation may be considered as a remedial measure in the presence of total or nearly total fertilization failure in ICSI.

关 键 词：人工激活 胚胎 性染色体 IGF-Ⅱ 基因表达 钙离子载体A23187 嘌呤霉素 人工受精 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]