小鼠精子和球形精子细胞显微注射受精研究

小鼠精子和球形精子细胞显微注射受精研究

小鼠精子和球形精子细胞显微注射受精研究

解剖学报 1999年第2期第0卷论著

作者：李子义谭景和孙兴参刘忠华周琪贺桂馨

单位：李子义谭景和孙兴参刘忠华周琪贺桂馨东北农业大学生物

工程系，哈尔滨150030 关键词：精子；球形精子细胞；显微注射；电活化；电融合；小鼠

【摘要】目的研究动物受精机理。方法以昆明白鼠为实验动物，利用显微注射技术，对精子和球形精子细胞的显微注射受精进行了研究。结果1. 带下注射受精，注射4～6个精子的卵的受精率(32.5%)和卵裂率(25.0%)显著高于注射单个精子的卵的受精率和卵裂率(分别为12.5%和6.3%)(P＜0.05)。

2. 带下注射4～6精子后，有第1极体卵母细胞的存活率(69.6%)、受精率(32.5%)和卵裂率(25.0%)分别高于无第1极体卵母细胞的存活率、受精率和卵裂率(分别为64.4%、20.7%和1

3.8%)，但两者无显著差异(P＞0.05)。3.胞质内精子注射卵的存活率(21.4%)极显著低于带下注射卵的存活率(69.6%)(P＜

0.01)；而胞质内精子注射卵的受精率(35.5%)、卵裂率(32.3)%和囊胚率(20.0%)则高于带下注射卵的受精率(33.3%)、卵裂率(25.0%)和囊胚率(10.0%)，但无显著差异(P＞0.05)。4.将球形精子细胞注入经电活化处理的卵母细胞的透明带下，注射卵的存活率为90.5%，电融合率为34.6%，2-细胞分裂率为28.3%。

结论 1.注射精子的数量对小鼠带下注射的受精率和卵裂率有显著影响。2.卵母细胞中第1极体存在与否对小鼠带下注射的受精率和卵裂率无显著影响。3. 不同注射方法对小鼠卵的存活率有极显著的影响，而对受精率、卵裂率和囊胚

率无显著影响。4. 本实验所采用的电融合条件，尚未满足多数球形精子细胞与卵母细胞融合之需要。

FERTILIZATION BY MICROINJECTION OF SPERMATOZOON AND ROUND SPERMATID

INTO OOCYTE IN THE MOUSE

Li Ziyi，Tan Jinghe△， Sun Xingshen, Liu Zhonghuan, Zhou Qi, He Guixin

(Department of Biotechnology, Northeast Agricultural University)

【Abstract】 Objective To study fertilization mechanism in mammals and provide essential data for treating human male

infertility.Methods Using microinjection technique, the

fertilization by microinjection of spermatozoa and round spermatids into oocytes in the mouse was systematically studied.Results 1. Following subzonal injection of spermatozoa into occytes, the fertilization rate (32.5%) and cleavage rate(25.0%) in the oocytes injected with 4-6 spermatozoa were significantly higher (P＜0.05) than those (12.5% and 6.3%, respectively) in the oocytes injected with a single spermatozoon. 2. Following subzonal injection of 4-6 spermatozoa into the oocytes, the survival rate (69.6%),

fertilization rate (32.5%) and cleavage rate (25.0%) in those oocytes with polar body 1 (PB1) were higher than those (64.4%, 20.7% and

13.8%, respectively) in the oocytes without PB1, but the difference was not significant (P＞0.05). 3. The survival rate (21.4%) in the oocytes injected with a single spermatozoon into the cytoplasm was significantly lower (P＜0.01) than that (69.0%) in the oocytes injected with 4\|6 spermatozoa into the perivitelline space (PVS). The fertilization rate (35.5%), cleavage rate (32.2%) and blastocyst rate (20.0%) in those oocytes injected with a single spermatozoon

into the cytoplasm of oocytes were higher than those (33.3%, 25.0% and 10.0%, respectively) in the oocytes injected with 4-6 spermatozoa into the PVS, but they did not differ significantly(P＞0.05). 4. Following subzonal injection of a round spermatid into the oocyte,