An anti-actin monoclonal antibody inhibits the zona pellucida-induced acrosome reaction and hyperactivated motility of human sperm.
An anti-actin monoclonal antibody inhibits the zona pellucida-induced acrosome reaction and hyperactivated motility of human sperm.
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DOI：
10.1093/molehr/8.1.37
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发表时间：
2002
影响因子：
4
通讯作者：
D. Y. Liu;M. Martić;G. Clarke;I. Grković;C. Garrett;M. E. Dunlop;H. Baker
中科院分区：
文献类型：
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作者：
D. Y. Liu;M. Martić;G. Clarke;I. Grković;C. Garrett;M. E. Dunlop;H. Baker
我们报道了一种抗肌动蛋白单抗(MAb)对人透明带(ZP)诱导的顶体反应(AR)的抑制作用。将活动精子与天然人ZP在含有抗肌动蛋白单抗、无关对照单抗或细胞松弛素B或D的培养液中孵育2 h，细胞松弛素B或D(40微mol/L)。回收与ZP结合的精子，用荧光素标记的豌豆凝集素测定AR。用抗小鼠免疫球蛋白G(MIGG)、免疫荧光和免疫金标记法检测抗肌动蛋白单抗在精子中的定位。抗肌动蛋白单抗可显著抑制ZP诱导的AR(相当于细胞松弛素)、离子载体A23187诱导的AR和精子在培养液中的过度激活。与抗肌动蛋白单抗孵育后，与精子头部结合的抗-MIGG小球回收了50%的精子，10%的精子留在了培养液中。在有抗肌动蛋白单抗存在的情况下，精子从与ZP结合恢复后与抗MIGG珠子结合的比例与ZP在没有抗体的情况下诱导的AR显著相关。免疫荧光和免疫胶体金法证实抗肌动蛋白单抗进入精子。本研究提示，在获能和AR启动过程中，精子质膜对抗肌动蛋白单抗具有通透性。
We report an inhibitory effect of an anti-actin monoclonal antibody (mAb) on the human zona pellucida (ZP)-induced acrosome reaction (AR). Motile sperm were incubated with native human ZP for 2 h in medium containing either the anti-actin mAb, an irrelevant control mAb or cytochalasins B or D (40 micromol/l). Sperm bound to the ZP were recovered and the AR was determined by fluorescein-labelled Pisum Sativum agglutinin. Anti-mouse immunoglobulin G (mIgG) Dynabeads, immunofluorescence and immunogold were used to detect the location of the anti-actin mAb in sperm. The anti-actin mAb significantly inhibited the ZP-induced AR (equivalent to cytochalasins), the ionophore A23187-induced AR and hyperactivation of sperm in medium. After incubation with anti-actin mAb, anti-mIgG beads bound to the head of >50% of sperm recovered after binding to the ZP and 10% of sperm remaining in the medium. The proportion of sperm that bound anti-mIgG beads after recovery from binding to the ZP in the presence of the anti-actin mAb was significantly correlated with the ZP-induced AR in the absence of the antibody. Immunofluorescence and immunogold demonstrated entry of the anti-actin mAb into sperm. This study suggests that the sperm plasma membrane becomes permeable to the anti-actin mAb during capacitation and initiation of the AR.