Acrosome intact and acrosome-reacted human sperm can initiate binding to the zona pellucida.
Acrosome intact and acrosome-reacted human sperm can initiate binding to the zona pellucida.
复制标题
顶体完整和顶体反应的人类精子可以启动与透明带的结合。
DOI：
10.1016/0012-1606(89)90042-0
复制
发表时间：
1989
影响因子：
2.7
通讯作者：
Hanson,FW
中科院分区：
文献类型：
--
作者：
Morales,P;Cross,NL;Overstreet,JW;Hanson,FW
哺乳动物的精子在穿透透明带之前必须经过顶体反应。在一些物种中，精子在与透明带结合之前经历顶体反应，而在其他物种中，只有顶体完整的精子才能与透明带结合。在这项研究中，我们解决了顶体状态和精子带与人类配子结合的问题。用人卵泡液n -（6-氨基己基）-5-氯萘磺酰胺（W-7）诱导精子顶体反应。含有不同百分比顶体反应精子的精子悬液与人卵母细胞孵育1分钟。无论诱导顶体反应的处理方式如何，与带结合的精子群体顶体状态与悬浮精子群体顶体状态相似（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可以启动与透明带的结合。然而，我们早前报道（N. L. Cross， P. Morales， J. W. Overstreet, and F. W. Hanson, 1988,Biol）。（第38,235 - 244号论文）表明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（该培养基中添加srcl2和精胺以支持精子运动和带结合）。在0 Ca-mTyr中，顶体反应精子在带上的比例仍然与悬浮中反应精子的比例高度相关，表明精子在结合前发生了反应。证明0 - Ca-mTyr有效抑制透明带诱导的顶体反应的证据来自这样的实验：用人卵泡液或对照培养基处理精子，并用0 - Ca-mTyr或对照培养基稀释悬浮液。加入人卵母细胞1分钟（脉冲），固定部分卵母细胞，其余卵母细胞转移至无精子培养基中孵育35分钟（追逐）后固定。在对照培养基中稀释的精子，用人类卵泡液或对照培养基预处理，在追逐后，带结合精子中顶体反应的百分比也有类似的增加（40%）。在0ca - mtyr中稀释的精子在追逐后没有显示顶体反应精子在透明带上的百分比增加。无论精子是否用人类卵泡液或对照培养基预处理，都是如此。这些观察结果表明，完整顶体和顶体反应的人类精子与透明带结合的效率相同。
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 orN-(6-aminohexyl)-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 SrCl2and 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. 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. This was the case whether the sperm had been pretreated with human follicular fluid or control medium. These observations indicate that acrosome intact and acrosome-reacted human sperm bind with equal efficiency to the zona pellucida.