ACROSOME STATUS AND MORPHOLOGY OF HUMAN SPERMATOZOA BOUND TO THE ZONA-PELLUCIDA AND OOLEMMA DETERMINED USING OOCYTES THAT FAILED TO FERTILIZE IN-VITRO
ACROSOME STATUS AND MORPHOLOGY OF HUMAN SPERMATOZOA BOUND TO THE ZONA-PELLUCIDA AND OOLEMMA DETERMINED USING OOCYTES THAT FAILED TO FERTILIZE IN-VITRO
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DOI：
10.1093/oxfordjournals.humrep.a138570
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发表时间：
1994-04-01
影响因子：
6.1
通讯作者：
BAKER, HWG
中科院分区：
文献类型：
--
作者：
LIU, DY;BAKER, HWG
在透明带完整的卵母细胞正常受精过程中，只有顶体反应的精子才能与卵膜融合。本研究的目的是确定人类无透明带卵母细胞的胚膜是否选择性地结合具有正常形态的精子和反应顶体。由于严重的精子缺陷而无法在体外受精的卵母细胞被使用。用酸性（pH 2-3）生理盐水去除透明带。精子样本取自正常生育能力的捐献者和无精子男性。游动精子选择游动精子，与卵母细胞孵育2 × 10(6)/ml。23个实验的结果表明，在2 h时，顶体反应精子与透明带结合的平均百分比（平均+/- SD， 42 +/- 22）显著高于受精培养基（27 +/- 12）。与此相反，所有与胚轴结合的精子在2 h时都发生顶体反应。此外，每个新鲜透明带的卵母细胞有100个精子结合，而每个无透明带的卵母细胞平均有28个精子结合（范围4-81）。新鲜透明带（45 +/- 21）和盐储存透明带（35 +/- 22）诱导的顶体反应无显著差异。与透明带结合的精子形态正常的百分率（84 +/- 13）和无透明带卵母细胞的卵膜形态正常的百分率（71 +/- 25）极显著高于受精介质中精子形态正常的百分率（39 +/- 11）（P < 0.01）。延长精子与透明带完整卵母细胞孵育的时间，发生顶体反应的精子比例增加（n = 6、2 h, 41 +/- 23; 3 h, 53 +/- 31; 4 h, 61 +/- 34）。然而，在单个精子样本之间，与透明带结合的精子之间顶体反应的百分比差异很大。综上所述，只有顶体反应的精子才能与无透明带卵母细胞的卵膜结合。新鲜和盐储存的人透明带菌在诱导顶体反应方面同样有效。无透明带卵母细胞的透明带和卵膜都能选择性地与正常形态的精子结合。
It is known that only acrosome-reacted spermatozoa can fuse with the oolemma during normal fertilization with zona pellucida-intact oocytes. The aim of this study was to determine if the oolemma of human zona pellucida-free oocytes selectively binds spermatozoa with normal morphology and a reacted acrosome. Oocytes that failed to fertilize in vitro because of severe sperm defects were used. The zona pellucida was removed with acidic (pH 2-3) saline. Sperm samples were obtained from normal fertile donors and normozoospermic men. Motile spermatozoa were selected with a swim-up technique and 2 x 10(6)/ml incubated with oocytes. The results from 23 experiments showed that at 2 h there was a significantly higher mean percentage of acrosome-reacted spermatozoa bound to the zona pellucida (mean +/- SD, 42 +/- 22) than in the insemination medium (27 +/- 12). In contrast, all spermatozoa bound to the oolemma at 2 h were acrosome reacted. Furthermore, each fresh zona pellucida had > 100 spermatozoa bound compared with an average of 28 (range 4-81) spermatozoa bound per zona pellucida-free oocyte. There was no significant difference in the zona pellucida-induced acrosome reaction between fresh (45 +/- 21) and salt-stored (35 +/- 22) zonae pellucidae. The percentage with normal morphology was significantly higher for spermatozoa bound to the zona pellucida (84 +/- 13) and oolemma of zona pellucida-free oocytes (71 +/- 25) than for spermatozoa in insemination medium (39 +/- 11) (P < 0.01). Extending the time of incubation of spermatozoa with zona pellucida-intact oocytes increased the proportion of spermatozoa undergoing the acrosome reaction (n = 6, 2 h, 41 +/- 23; 3 h, 53 +/- 31; 4 h, 61 +/- 34). However, there was a large variation in the percentage of acrosome reactions among spermatozoa bound to the zona pellucida between individual sperm samples. In conclusion, only acrosome-reacted spermatozoa can bind to the oolemma of zona pellucida-free oocytes. Both fresh and salt-stored human zonae pellucidae are equally effective in inducing the acrosome reaction. Both the zona pellucida and the oolemma of zona pellucida-free oocytes selectively bind spermatozoa with normal morphology.