Saxena D K, Tanii I, Oh-oka T, Yoshinaga K, Toshimori K
Department of Anatomy and Reproductive Cell Biology, Miyazaki Medical College, Kiyotake, Japan.
Zygote. 2000 Nov;8(4):329-38. doi: 10.1017/s096719940000112x.
In this study we examined the behaviour and role of an intra-acrosomal antigenic molecule, acrin 3, during mouse fertilisation in vitro by assessing the effect of its pertinent monoclonal antibody mMC101. Experiments were designed to assess the effect of mMC101 on sperm-zona pellucida binding, the acrosome reaction, zona pellucida penetration, sperm-egg fusion, and fertilisation in vitro. mMC101 did not affect sperm motility or primary and secondary binding to the zona pellucida, but significantly inhibited fertilisation of zona-pellucida-intact oocytes in a dose-dependent manner. In the presence of mMC101 at 100 microg/ml concentration in TYH medium, none of the oocytes developed to pronuclear stage by 5 h after co-incubation of the gametes, but the pronucleus formation rate recovered to some extent (45.3%) after 8 h, indicating a delay of early embryonic development. mMC101 also delayed and significantly suppressed zona pellucida penetration by sperm. Acrin 3 dispersed and did not remain on completely acrosome-reacted sperm. Although mMC101 did not influence the zona-pellucida-induced acrosome reaction, it significantly inhibited fertilisation when acrosome-reacted sperm in the presence of mMC101 inseminated zona-pellucida-free oocytes. However, fertilisation remained unaffected when acrosome-reacted sperm in the absence of mMC101 inseminated zona-pellucida-free oocytes even in its presence. Thus, acrin 3 appears to facilitate zona pellucida penetration and is also likely to be involved in sperm-oocyte fusion by modifying the sperm plasma membrane during the acrosome reaction.
在本研究中,我们通过评估其相关单克隆抗体mMC101的作用,研究了顶体内抗原分子acrin 3在小鼠体外受精过程中的行为和作用。实验旨在评估mMC101对精子与透明带结合、顶体反应、透明带穿透、精卵融合及体外受精的影响。mMC101不影响精子活力以及与透明带的初次和二次结合,但以剂量依赖方式显著抑制完整透明带卵母细胞的受精。在TYH培养基中存在浓度为100μg/ml的mMC101时,配子共孵育5小时后,没有卵母细胞发育到原核阶段,但8小时后原核形成率有所恢复(45.3%),表明早期胚胎发育延迟。mMC101还延迟并显著抑制精子对透明带的穿透。Acrin 3分散且不会保留在完全发生顶体反应的精子上。虽然mMC101不影响透明带诱导的顶体反应,但当存在mMC101的顶体反应精子使无透明带卵母细胞受精时,它会显著抑制受精。然而,即使存在mMC101,当不存在mMC101的顶体反应精子使无透明带卵母细胞受精时,受精仍不受影响。因此,acrin 3似乎有助于透明带穿透,并且也可能通过在顶体反应过程中修饰精子质膜而参与精卵融合。