Alavi Sayyed Mohammad Hadi, Postlerová-Maňásková Pavla, Hatef Azadeh, Pšenička Martin, Pěknicová Jana, Inaba Kazuo, Ciereszko Andrzej, Linhart Otomar
Faculty of Fisheries and Protection of Waters, Research Institute of Fish Culture and Hydrobiology, University of South Bohemia, 389 25, Vodňany, Czech Republic,
Fish Physiol Biochem. 2014 Oct;40(5):1393-8. doi: 10.1007/s10695-014-9933-8. Epub 2014 Mar 28.
In mammals, proteases are present in sperm acrosome and play key role in fertilization. Sturgeon sperm has an acrosome, but its physiology, biochemistry, and potential role in fertilization are unknown. In the present study, we have observed high protease activity in acidic extract of intact sperm compared to that of seminal plasma in sterlet (Acipenser ruthenus). The protease activity was decreased and increased in acidic extract of motility-activated sperm and in the activation medium, respectively. Molecular analysis revealed total protease and serine (acrosin) protease activities in sperm acidic extract which was accumulated in a protein band with relative molecular mass of 35 kDa. Immunoelectron microscopy using an affinity-purified polyclonal antibody for boar acrosin localized the protease at the acrosome region. Moreover, initiation of sperm motility was inhibited after activation in the presence of inhibitors for both trypsin-like and chymotrypsin-like proteases, while the effects of protease inhibitors on sperm velocity were uncertain. Our results indicate similarities in physiology and biochemistry of acrosome between sturgeon and mammals and suggest potential role of protease in the initiation of sperm motility in sturgeon.
在哺乳动物中,蛋白酶存在于精子顶体中,并在受精过程中发挥关键作用。鲟鱼精子有一个顶体,但其生理学、生物化学以及在受精中的潜在作用尚不清楚。在本研究中,我们观察到,与小体鲟(Acipenser ruthenus)精浆相比,完整精子酸性提取物中的蛋白酶活性较高。蛋白酶活性在运动激活精子的酸性提取物和激活培养基中分别降低和升高。分子分析显示精子酸性提取物中存在总蛋白酶和丝氨酸(顶体蛋白酶)蛋白酶活性,这些活性积聚在一条相对分子质量为35 kDa的蛋白带中。使用针对猪顶体蛋白酶的亲和纯化多克隆抗体进行免疫电子显微镜观察,将蛋白酶定位在顶体区域。此外,在存在胰蛋白酶样和糜蛋白酶样蛋白酶抑制剂的情况下激活后,精子运动的启动受到抑制,而蛋白酶抑制剂对精子速度的影响尚不确定。我们的结果表明鲟鱼和哺乳动物顶体在生理学和生物化学上具有相似性,并提示蛋白酶在鲟鱼精子运动启动中的潜在作用。