Yoshida Kaoru, Krasznai Zoárd Tibor, Krasznai Zoltán, Yoshiike Miki, Kawano Natsuko, Yoshida Manabu, Morisawa Masaaki, Tóth Zoltán, Bazsáné Zsuzsa Kassai, Márián Teréz, Iwamoto Teruaki
Biomedical Engineering Center, Toin University of Yokohama, 1614 Kurogane-cho, Aoba-ku, Yokohama, Japan.
Cell Motil Cytoskeleton. 2009 Feb;66(2):99-108. doi: 10.1002/cm.20329.
Semenogelin I and II (Sgs) are the major component of human semen coagulum. The protein is rapidly cleaved after ejaculation by a prostate-specific antigen, resulting in liquefaction of the semen coagulum and the progressive release of motile spermatozoa. Sgs inhibit human sperm motility; however, there is currently no information on its effect on the sperm membrane. This study investigated the role of Sgs on human sperm motility through regulation of membrane potential and membrane permeability. Fresh semen samples were obtained from normozoospermic volunteers, and studies were conducted using motile cells selected using the swim-up method. Sgs changed the characteristics of sperm motion from circular to straightforward as evaluated by a computer-assisted motility analyzer, and all parameters were decreased more than 2.5 mg/mL. The results demonstrate that Sgs treatment immediately hyperpolarized the membrane potential of swim-up-selected sperm, changed the membrane structure, and time-dependently increased membrane permeability, as determined through flow cytometric analysis. The biphasic effects of Sgs were time- and dose-dependent and partially reversible. In addition, a monoclonal antibody against Sgs showed positive binding to cell membrane proteins in fixed cells, observed with confocal fluorescence microscopy. These results demonstrate that Sgs modifies the membrane structure, indirectly inhibiting motility, and provides suggestions for a therapy for male infertility through selection of a functional sperm population using Sgs.
精液凝固蛋白I和II(Sgs)是人类精液凝块的主要成分。射精后,该蛋白会被前列腺特异性抗原迅速裂解,导致精液凝块液化,并逐步释放出有活力的精子。Sgs会抑制人类精子的活力;然而,目前尚无关于其对精子膜影响的相关信息。本研究通过调节膜电位和膜通透性来探究Sgs在人类精子活力方面的作用。从正常精子志愿者那里获取新鲜精液样本,并使用上浮法筛选出有活力的细胞进行研究。通过计算机辅助精子活力分析仪评估,Sgs使精子运动特征从圆周运动变为直线运动,且当浓度超过2.5 mg/mL时所有参数均下降。结果表明,通过流式细胞术分析确定,Sgs处理会立即使上浮法筛选出的精子膜电位超极化,改变膜结构,并随时间增加膜通透性。Sgs的双相效应具有时间和剂量依赖性且部分可逆。此外,用共聚焦荧光显微镜观察发现,一种针对Sgs的单克隆抗体与固定细胞中的细胞膜蛋白呈阳性结合。这些结果表明,Sgs会改变膜结构,间接抑制精子活力,并为通过使用Sgs选择功能性精子群体来治疗男性不育症提供了建议。