Liu D Y, Garrett C, Baker H W G
University of Melbourne Department of Obstetrics and Gynaecology, Reproductive Services, Royal Women's Hospital and Melbourne IVF, Vic., Australia.
Int J Androl. 2006 Aug;29(4):475-81. doi: 10.1111/j.1365-2605.2006.00681.x. Epub 2006 Feb 15.
In the literature there is still confusion whether acrosome-reacted sperm in medium can initiate primary binding to human zona pellucida (ZP). The ability of acrosome-reacted sperm to bind to ZP in vitro can be deduced by measuring the acrosome reaction (AR) of ZP-bound sperm compared with sperm in medium after incubation under different conditions inhibiting the ZP-induced AR. Motile sperm from fertile men, normospermic men and infertile men diagnosed with disordered ZP-induced AR (DZPIAR) were selected by swim-up (2 x 10(6) in 1 mL medium) and incubated for 1-2 h with four oocytes from failed in vitro fertilization (IVF). The acrosome status of sperm was assessed using pisum sativum agglutinin labelled with fluorescein. The ZP-induced AR was inhibited in experiments using sperm from DZPIAR patients, hyper-osmotic medium (400 mOsm/kg) and medium containing soybean trypsin inhibitor (SBTI; 4 mg/mL). Pre-treatment with calcium ionophore was used to create a sperm population with elevated AR. In all experiments with factors inhibiting the ZP-induced AR, the AR was significantly lower for ZP-bound sperm compared with sperm in medium: DZPIAR patients 4% vs. 15%, hyper-osmotic medium 3% vs. 12%, SBTI 2% vs. 12% and SBTI 3% vs. 23% after treatment with calcium ionophore. In conclusion, acrosome-reacted sperm in vitro have significantly reduced, in fact probably zero ability to bind to the ZP.
在文献中，对于介质中顶体反应的精子是否能启动与人类透明带（ZP）的初次结合仍存在困惑。通过测量与在不同抑制ZP诱导的顶体反应（AR）条件下孵育后的介质中的精子相比，与ZP结合的精子的顶体反应（AR），可以推断顶体反应的精子在体外与ZP结合的能力。通过上游法（在1 mL介质中2×10⁶个）从生育能力正常的男性、精子正常的男性以及被诊断为ZP诱导的顶体反应紊乱（DZPIAR）的不育男性中选择活动精子，并与来自体外受精（IVF）失败的四个卵母细胞一起孵育1 - 2小时。使用荧光素标记的豌豆凝集素评估精子的顶体状态。在使用来自DZPIAR患者的精子、高渗介质（400 mOsm/kg）和含有大豆胰蛋白酶抑制剂（SBTI；4 mg/mL）的介质的实验中，ZP诱导的AR受到抑制。用钙离子载体进行预处理以产生AR升高的精子群体。在所有使用抑制ZP诱导的AR的因素的实验中，与介质中的精子相比，与ZP结合的精子的AR显著降低：DZPIAR患者为4%对15%，高渗介质为3%对12%，SBTI为2%对12%，以及用钙离子载体处理后SBTI为3%对23%。总之，体外顶体反应的精子与ZP结合的能力显著降低，实际上可能为零。
