Mettler L, Kim S K, Kuranty A
Department of Obstetrics and Gynecology, University of Kiel, Germany.
J Reprod Med. 1995 Dec;40(12):839-44.
To evaluate the outcome of a single sperm injection into the perivitelline space of bovine oocytes.
Oocytes obtained from the ovaries of slaughtered heifers and cows were cultured in medium 199 supplemented with 20% fetal calf serum for 24 hours. Fresh or frozen spermatozoa were incubated for two hours in Ham's F-10 medium containing 0.75% bovine serum albumin for capacitation and kept for 30 minutes in culture medium containing 12 mM dibutyryl cyclic guanosine 3',5' monophosphate and 10 mM imridazole for the acrosome reaction. Only one motile spermatozoon was injected into the perivitelline space of each oocyte.
The second polar body and the pronuclear stages were observed in 9.5% and 5.4% of oocytes, respectively. The cleavage rate of the oocytes over the two-cell stage was 4.1% (10 of 242).
The results of this study suggest that microinjection may be a useful technique for studying sperm-oocyte interaction. However, one spermatozoon is not enough to achieve an acceptable fertilization rate.
评估向牛卵母细胞透明带周隙内单精子注射的结果。
从屠宰的小母牛和母牛卵巢获取的卵母细胞，在添加20%胎牛血清的199培养基中培养24小时。新鲜或冷冻的精子在含0.75%牛血清白蛋白的哈姆F-10培养基中孵育两小时以进行获能，然后在含12 mM二丁酰环鸟苷3',5'-单磷酸和10 mM咪唑的培养基中保存30分钟以诱导顶体反应。向每个卵母细胞的透明带周隙内仅注射一个活动精子。
分别在9.5%和5.4%的卵母细胞中观察到第二极体和原核期。两细胞期以上卵母细胞的分裂率为4.1%（242个中有10个）。
本研究结果表明，显微注射可能是研究精卵相互作用的一种有用技术。然而，一个精子不足以实现可接受的受精率。
