精子脱尾温育降低小鼠卵胞浆内单精子显微注射的受精率  

Tail cutting-off and incubation of sperm reduce fertilization of ICSI in mouse

作　　者：苗聪秀[1] 燕志光[2] 杨红梅[1] 龙慧[3] 梁红星[2] 于莎[4] 匡延平[3] 吕祁峰[3] 

机构地区：[1]长治医学院附属和平医院生殖医学中心,山西长治046000 [2]亚热带农业生物资源保护与利用国家重点实验室(广西大学动物繁殖研究所),广西南宁530004 [3]上海交通大学医学院附属第九人民医院辅助生殖科,上海200011 [4]上海中医药大学附属曙光医院妇科,上海200021

出　　处：《中国现代医学杂志》2013年第14期1-4,共4页China Journal of Modern Medicine

基　　金：国家自然科学基金(No:31071275);国家自然科学基金青年项目(No:31101070);上海市科委项目(No:09411962900;No:12ZR1416600)

摘　　要：目的脱尾的精子头注射是小鼠显微授精的一种常用方法,脱尾对精子造成的膜损伤是否影响卵胞浆内单精子显微注射(ICSI)的受精率和卵裂率尚不清楚,该研究拟初步探索精子的机械脱尾等膜损伤以及不同温育条件对ICSI的受精率和卵裂率的影响。方法首先,用机械吹打法处理小鼠精子悬液,使得部分精子脱尾,于不同温度下孵育,观察孵育温度对ICSI的受精率和卵裂率的影响;然后比较精子化学破膜与吹打脱尾处理后温育一定时间对显微授精的受精率、卵裂率的影响。结果相对于对照组,对小鼠精子进行吹打脱尾,经37℃下温育12 h后的显微授精双原核形成率和卵裂率显著下降(75.6%vs 15.1%,P<0.01;91.1%vs 18.9%,P<0.01),而4℃下温育12 h的结果未见正常受精率和卵裂率受显著影响。用Triton-100精子进行化学破膜后温育,尽管其没有机械脱尾法效果显著,也同样明显降低了受精率和卵裂率(P<0.01)。结论该研究初步提示,精子脱尾等破膜损伤在经37℃较长时间温育后会降低小鼠ICSI的受精率,常规小鼠ICSI操作中小鼠精子批量脱尾处理后不宜在37℃下放置过久。【Objective】 Intracytoplasmic sperm injection with isolated sperm head is accepted widely in assisted reproduction techniques.However,the effect of membrane damage induced by tail cutting off or others on the fertilization and cleavage of ICSI is unclear.The current primary research focuses on the effects of membrane damage induced by mechanical tail cutting off or others followed by different incubation conditions on the fertilization and cleavage of ICSI.【Methods】 First,mechanical blowing was applied to isolate sperm heads from tails,then incubating sperm heads at different degree before intracytoplasmic sperm injection.The relationship between the fertilization and cleavage and the incubation conditions was analyzed.Furthermore,the effects of incubation after chemical or mechanical damage of sperm membrame on the rates of fertilization and cleavage were compared.【Results】 Compared the normal control,when sperms were subjected to mechanical tail cutting off and incubation at 37℃ for 12 hours,the rates of 2 prenuclear formation and cleavage decreased significantly（75.6% vs 15.1%,P〈 0.01;and 91.1% vs 18.9%,P〈 0.01）,however,while the incubation was at 4℃ for 12 h,the results were not affected significantly.After chemical damage of sperm membrane with Triton-100,both fertilization and cleavage decreased significantly（P〈 0.01）,although it showed less reduction than the tail cutting off.【Conclusion】 Our primary data suggested that long time incubation at 37℃ of sperm following membrane damage,such as tail-cutting off,affects the fertilization rate of mouse ICSI,so,the sperms following tail cutting off in mouse ICSI should not be long time cultured at 37℃.

关 键 词：卵胞浆内单精子显微注射(ICSI) 受精率 卵裂率 脱尾 小鼠 

分 类 号：R711.6[医药卫生—妇产科学]