Birck Anders, Labouriau Rodrigo, Christensen Preben
Copenhagen University, Faculty of Life Sciences, Department of Large Animal Sciences, Veterinary Reproduction and Obstetrics, Frederiksberg, Denmark.
Anim Reprod Sci. 2009 Oct;115(1-4):124-36. doi: 10.1016/j.anireprosci.2008.10.017. Epub 2008 Oct 31.
The present study investigated the dynamics of the in vitro induced acrosome reaction (AR) in boar sperm in response to medium composition, incubation time and ionophore concentration. The AR is a prerequisite for normal sperm fertilizing capability and can be studied in vitro following induction by various agents. The ability of a sperm population to undergo the AR in vivo is expected to influence male fertilizing potential, and attempts to relate the in vitro induced AR to fertility has been reported. However, to relate the induced AR to fertility one should be aware of the dynamics of the in vitro induced AR. A detailed description of the dynamics of sperm viability and acrosomal status of boar sperm following in vitro induction of the AR has to our knowledge not previously been conducted. In the present study, a triple color flow cytometric detection technique was used, which gave simultaneous information on sperm viability and acrosomal status. The ionophore induced AR was dependent on extracellular Ca(2+), but could be easily induced in boar sperm without capacitation. Capacitation-associated plasma membrane phospholipid scrambling was assessed and a medium specific ability to induce these membrane changes was observed. Both sperm viability and the induced AR were significantly affected by sperm capacitation, incubation time and ionophore concentration. The results lead to suggestions for an optimized AR induction protocol that takes both sperm viability and the effectiveness of AR induction into consideration.
本研究调查了体外诱导公猪精子顶体反应（AR）的动力学，该反应受培养基成分、孵育时间和离子载体浓度的影响。顶体反应是精子正常受精能力的先决条件，可在体外通过各种试剂诱导后进行研究。精子群体在体内发生顶体反应的能力有望影响雄性受精潜力，并且已有报道尝试将体外诱导的顶体反应与生育力联系起来。然而，要将诱导的顶体反应与生育力联系起来，应该了解体外诱导顶体反应的动力学。据我们所知，此前尚未对体外诱导顶体反应后公猪精子的活力和顶体状态的动力学进行详细描述。在本研究中，使用了三色流式细胞术检测技术，该技术可同时提供精子活力和顶体状态的信息。离子载体诱导的顶体反应依赖于细胞外Ca(2+)，但在未获能的公猪精子中也很容易诱导。评估了与获能相关的质膜磷脂 scrambling，并观察到培养基诱导这些膜变化的特定能力。精子活力和诱导的顶体反应均受精子获能、孵育时间和离子载体浓度的显著影响。结果提出了一种优化的顶体反应诱导方案，该方案同时考虑了精子活力和顶体反应诱导的有效性。
