Mouse sperm antigens that participate in fertilization. IV. A monoclonal antibody prevents zona penetration by inhibition of the acrosome reaction.
Mouse sperm antigens that participate in fertilization. IV. A monoclonal antibody prevents zona penetration by inhibition of the acrosome reaction.
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参与受精的小鼠精子抗原。
DOI：
10.1016/0012-1606(86)90318-0
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发表时间：
1986
影响因子：
2.7
通讯作者：
Saling,PM
中科院分区：
文献类型：
--
作者：
Saling,PM
为了研究哺乳动物配子相互作用的分子基础，用小鼠睾丸同基因免疫制备了单克隆抗体。先前的工作已经描述了两种特定的单克隆抗体，M41和M42，它们不可分割地定位于覆盖顶体的限制性部分的质膜，但识别不同的抗原。其中一种单克隆抗体M42在体外对小鼠受精有显著的抑制作用，但仅在透明质酸存在的情况下，而M41对受精过程中的任何可测定事件都没有明显的影响。进行此处描述的实验以确定受M42 mAb影响的精子-精子相互作用的精确事件（精子-精子结合、诱导顶体反应或穿透精子）。与使用无mAb或M41 mAb处理的对照精子达到的水平相比，用M42 mAb预处理后获能小鼠精子与透明带的结合未减弱。当检查mAb对透明带诱导的AR的影响时，精子表面顶体反应（AR）精子的百分比随时间增加，在授精后约90 min达到平台，对照组和M41 mAb处理组中有78%的结合细胞AR。M42处理的精子在测定的120分钟间隔内从未达到大于23%的AR细胞。为了定量这种影响，获能精子暴露于浓度增加的酸溶解的透明带。在对照组和M41 mAb给药组中发现AR精子比例增加，最高达70-76% AR细胞，8或12个透明带/μl。相比之下，M42处理的精子在测试的整个透明带浓度范围内仅显示21-28%的AR细胞。当用A23187诱导顶体反应时，出现了完全不同的结果：M42不再能够防止AR。A23187能够超越M42 mAb对AR的抑制作用，因此可以特异性检查M42 mAb对精子穿透透明膜的可能影响。在存在A23187的情况下，M42 mAb处理的精子的精子穿透水平在定性和定量方面均与相同条件下的对照和M41 mAb处理的精子相当。因此，似乎M42 mAb鉴定了小鼠精子的高分子量双联体（220-240 kDa），其特异性参与生理条件下发生的精子顶体反应的诱导。
To investigate the molecular basis of gamete interaction in mammals, monoclonal antibodies (mAbs) have been generated by syngeneic immunization with mouse testis. Previous work has described two particular mAbs, M41 and M42, which localize indistinguishably to the plasma membrane overlying a restricted portion of the acrosome, but recognize different antigens. One of the mAbs, M42, inhibits mouse fertilizationin vitrosignificantly, but only in the presence of the zona pellucida, whereas M41 has no apparent effect upon any assayable event in the fertilization process. The experiments described here were performed to identify the precise event of sperm-zona interaction (sperm-zona binding, induction of the acrosome reaction, or penetration through the zona) that is affected by M42 mAb. Capacitated mouse sperm binding to the zona pellucida was undiminished following pretreatment with M42 mAb, when compared to levels achieved using either no mAb- or to M41 mAb-treated control sperm. When the effect of mAbs on the zona-induced AR was examined, the percentage of acrosome reacted (AR) sperm at the zona surface increased with time, plateauing at approximately 90 min post-insemination, with 78% of the bound cells AR in the control and the M41 mAb-treated groups. M42-treated sperm never achieved greater than 23% AR cells over the 120-min interval assayed. To quantitate this effect, capacitated sperm were exposed to increasing concentrations of acid-solubilized zonae. Increased proportions of AR sperm were found in the control and M41 mAb-treated groups, up to a maximum of 70–76% AR cells with 8 or 12 zonae/μl. In contrast, M42-treated sperm displayed only 21–28% AR cells over the entire range of zonae concentrations tested. An entirely different result emerged when acrosome reactions were induced with A23187: M42 was no longer able to prevent the AR. This ability of A23187 to override M42 mAb's inhibitory effect on the AR permitted specific examination of the possible effect of M42 mAb on sperm penetration through the zona pellucida. In the presence of A23187, zona penetration levels for M42 mAb-treated sperm were equivalent, both qualitatively and quantitatively, to control and to M41 mAb-treated sperm under the same conditions. It appears, therefore, that M42 mAb identifies a high molecular weight doublet (220–240 kDa) of mouse sperm that participates specifically in the induction of the sperm's acrosome reaction as it occurs under physiological conditions.