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一种抗肌动蛋白单克隆抗体可抑制透明带诱导的人类精子顶体反应和超激活运动。

An anti-actin monoclonal antibody inhibits the zona pellucida-induced acrosome reaction and hyperactivated motility of human sperm.

作者信息

Liu D Y, Martic M, Clarke G N, Grkovic I, Garrett C, Dunlop M E, Baker H W G

机构信息

Department of Obstetrics and Gynecology, Anatomy and Cell Biology and Medicine, University of Melbourne, Royal Women's Hospital, Melbourne, Australia.

出版信息

Mol Hum Reprod. 2002 Jan;8(1):37-47. doi: 10.1093/molehr/8.1.37.

Abstract

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.

摘要

我们报道了一种抗肌动蛋白单克隆抗体(mAb)对人透明带(ZP)诱导的顶体反应(AR)具有抑制作用。将活动精子在含有抗肌动蛋白mAb、无关对照mAb或细胞松弛素B或D(40微摩尔/升)的培养基中与天然人ZP一起孵育2小时。回收与ZP结合的精子,并通过荧光素标记的豌豆凝集素测定AR。使用抗小鼠免疫球蛋白G(mIgG)磁珠、免疫荧光和免疫金来检测抗肌动蛋白mAb在精子中的位置。抗肌动蛋白mAb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以及培养基中精子的超活化。与抗肌动蛋白mAb孵育后,抗mIgG磁珠与>50%与ZP结合后回收的精子头部以及培养基中剩余的10%精子结合。在抗肌动蛋白mAb存在的情况下,从与ZP结合后回收的精子中结合抗mIgG磁珠的精子比例与不存在该抗体时ZP诱导的AR显著相关。免疫荧光和免疫金显示抗肌动蛋白mAb进入精子。这项研究表明,在获能和AR启动过程中,精子质膜对抗肌动蛋白mAb变得具有通透性。

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