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顶体完整和顶体反应的人类精子均可启动与透明带的结合。

Acrosome intact and acrosome-reacted human sperm can initiate binding to the zona pellucida.

作者信息

Morales P, Cross N L, Overstreet J W, Hanson F W

机构信息

Department of Obstetrics and Gynecology, School of Medicine, University of California, Davis 95616.

出版信息

Dev Biol. 1989 Jun;133(2):385-92. doi: 10.1016/0012-1606(89)90042-0.

Abstract

Mammalian sperm must be acrosome reacted before penetrating the zona pellucida. In some species the sperm undergo the acrosome reaction before binding to the zona pellucida and in other species only acrosome intact sperm can initiate binding to the zona. In this study we addressed the question of acrosomal status and sperm-zona binding with human gametes. Sperm acrosome reactions were induced by treatment with human follicular fluid or N-(6-amino-hexyl)-5-chloro-naphthalene sulfonamide (W-7). The sperm suspensions, containing various percentages of acrosome-reacted sperm, were then incubated with human oocytes for 1 min. The acrosomal status of the sperm population bound to the zona was similar to the acrosomal status of the population of sperm in suspension (R2 = 0.77), regardless of the treatment to induce acrosome reactions. Our interpretation of these results is that both acrosome intact and acrosome-reacted human sperm can initiate binding to the zona pellucida. However, we reported earlier (N. L. Cross, P. Morales, J. W. Overstreet, and F. W. Hanson, 1988, Biol. Reprod. 38, 235-244) that the human zona pellucida is able to induce acrosome reactions. Thus, to exclude the possibility that sperm had undergone the acrosome reaction on the zona within 1 min of binding, sperm were suspended in a nominally calcium-free Tyrode's medium (0 Ca-mTyr) before incubation with oocytes (this medium was supplemented with SrCl2 and spermine to support sperm motility and zona binding). In 0 Ca-mTyr, the proportion of acrosome-reacted sperm on the zona was still highly correlated with the proportion of reacted sperm in suspension, indicating that the sperm were reacted before binding. Evidence that 0 Ca-mTyr effectively inhibited acrosome reactions induced by the zona pellucida was derived from experiments in which sperm were treated with human follicular fluid or control medium and the suspensions were diluted with either 0 Ca-mTyr or control medium.4+ Human oocytes were added for 1 min (pulse) at which time some oocytes were fixed and other oocytes were transferred to sperm-free medium and incubated for 35 min (chase) before fixation. Sperm diluted in control medium, pretreated with either human follicular fluid or control medium, showed a similar increase (40%) in the percentage of acrosome reactions among the zona-bound sperm after the chase. Sperm diluted in 0 Ca-mTyr did not show an increase in the percentage of acrosome-reacted sperm on the zona pellucida after the chase.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

哺乳动物的精子在穿透透明带之前必须发生顶体反应。在某些物种中,精子在与透明带结合之前就发生顶体反应,而在其他物种中,只有顶体完整的精子才能启动与透明带的结合。在本研究中,我们探讨了人类配子的顶体状态与精子 - 透明带结合的问题。用人类卵泡液或N -(6 - 氨基己基)-5 - 氯萘磺酰胺(W - 7)处理诱导精子顶体反应。然后将含有不同百分比顶体反应精子的精子悬液与人类卵母细胞孵育1分钟。无论诱导顶体反应的处理方式如何,与透明带结合的精子群体的顶体状态与悬液中精子群体的顶体状态相似(R2 = 0.77)。我们对这些结果的解释是,顶体完整和顶体反应的人类精子都能启动与透明带的结合。然而,我们之前报道过(N. L. Cross、P. Morales、J. W. Overstreet和F. W. Hanson,1988年,《生物繁殖》38卷,235 - 244页)人类透明带能够诱导顶体反应。因此,为了排除精子在结合后1分钟内在透明带上发生顶体反应的可能性,在与卵母细胞孵育之前,将精子悬浮在名义上无钙的Tyrode培养基(0 Ca - mTyr)中(该培养基添加了SrCl2和精胺以支持精子活力和与透明带的结合)。在0 Ca - mTyr中,透明带上顶体反应精子的比例与悬液中反应精子的比例仍高度相关,表明精子在结合之前就已发生反应。0 Ca - mTyr有效抑制透明带诱导的顶体反应的证据来自以下实验:精子用人类卵泡液或对照培养基处理,然后将悬液用0 Ca - mTyr或对照培养基稀释。加入人类卵母细胞1分钟(脉冲),此时一些卵母细胞固定,其他卵母细胞转移到无精子培养基中并孵育35分钟(追踪)后再固定。用人类卵泡液或对照培养基预处理、在对照培养基中稀释的精子,在追踪后与透明带结合的精子中顶体反应的百分比有类似的增加(40%)。在0 Ca - mTyr中稀释的精子在追踪后透明带上顶体反应精子的百分比没有增加。(摘要截断于400字)

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