家兔显微受精影响因素及显微受精胚胎超微结构  

Affecting Factors of Microfertilization and Ultrastructure of Microfertilized Embryos in Rabbits

作　　者：李子义[1,2] 周琪[1,2] 邹啸环 文兴豪[1,2] 谭景和 

机构地区：[1]东北农业大学生物工程系 [2]解放军农牧大学

出　　处：《中国兽医学报》1997年第6期593-599,共7页Chinese Journal of Veterinary Science

基　　金：黑龙江省科委资助

摘　　要：以家兔为实验动物，利用胞质内精子注射技术和透明带下精子注射技术，对卵龄、精子获能、注射针管口径、操作室温和操作方法等影响显微受精的因素以及显微受精胚胎的超微结构进行了研究。结果显示：（１）胞质内单精子注射，ｈＣＧ后１５～１７ｈ卵的存活率为６９．４％，显著（Ｐ＜０．０５）高于ｈＣＧ后１９～２１ｈ卵的存活率（４７．３％）；而受精率和卵裂率（分别为５２．９％和４１．２％）分别低于ｈＣＧ后１９～２１ｈ卵的受精率（６５．４％）和卵裂率（４６．２％）。（２）将５～１０个在ＤＭ液中获能１２～１３ｈ的精子注入卵子的透明带下，ｈＣＧ后１９～２１ｈ卵的受精率（４９．２％）和卵裂率（４０．７％）分别显著（Ｐ＜０．０５）高于ｈＣＧ后１５～１７ｈ卵的受精率（２８．６％）和卵裂率（２２．５％）。（３）以ｍＨＩＳ１５ｍｉｎ＋ｍＤＭ３～４ｈ方法和ＨＩＳ１５ｍｉｎ＋ＤＭ５～６ｈ方法获能的精子带下注射的受精率（分别为６３．６％和５１．０％）和卵裂率（分别为５４．５％和４３．１％）分别显著（Ｐ＜０．０５）高于或略高于其他方法获能的精子带下注射的受精率和卵裂率。（４）内径７μｍ的注射针管用于胞质内单精子注射，卵的存活率（６９．４％）极显著（Ｐ＜０?The methods of intracytoplasmic sperm injection (ICSI) and subzonal injection (SUZI) were used to study the effects of egg age, sperm capacitation, inner diameter of injection pipette and room temperature on the microfertilization in rabbits, and the ultrastructure of microfertilized embryos of rabbits was also investigated. The results were as follows: 1. Following microinjection of a single spermatozoon into cytoplasm of the oocyte, the survival rate (69.4%) in oocytes collected 1517 h post hCG injection was significantly higher than that (47.3%) in oocytes collected 1921 h post hCG injection (P<0.05), but the fertilization rate (52.9%) and cleavage rate (41.2%) in oocytes collected 1517 h post hCG injection was lower than those (65.4% and 46.2%, respectively) in oocytes collected 1921 h post hCG injection (P>0.05). 2. The fertilization rate (49.2%) and cleavage rate (40.7%) in oocytes collected 1921 h post hCG injection were significantly (P<0.05) higher than those (28.6% and 22.5%, respectively) in oocytes collected 1517 h post hCG injection when 510 spermatozoa capacitated in DM for 1213 h had been microinjected into the perivitelline space (PVS) of the oocyte. 3. The fertilization rate and cleavage rate in oocytes injected with spermatozoa capacitated either in mHIS for 15 min and then mDM for 34 h or HIS for 15 min and then DM for 56 h were significantly higher (P<0.05) or slightly higher than those in oocytes injected with spermatozoa capacitated by other methods, when 510 spermatozoa microinjeced into the PVS of the oocyte. 4. The survival rate (69.4%), fertilization rate (52.9%) and cleavage rate (41.2%) in oocytes injected with an injection pipette with an inner diameter of 7 μm were significantly higher (P<0.01) than those (40.0%, 35.7% and 28.6%, respectively) in oocytes injected by injection pipettes with an inner diameter of 9 μm when a single spermatozoon was introduced into the cytoplasm of the oocyte. 5. The survival rate (69.4%),

关 键 词：显微受精 卵龄 精子获能 胚胎 超微结构 家兔 

分 类 号：S852.1[农业科学—基础兽医学] S829.1[农业科学—兽医学]