Nass S J, Miller D J, Winer M A, Ax R L
Department of Dairy Science, University of Wisconsin, Madison 53706.
Mol Reprod Dev. 1990 Mar;25(3):237-46. doi: 10.1002/mrd.1080250305.
Heparin binds to bovine sperm and stimulates capacitation in vitro. Seminal plasma alters the ability of epididymal sperm to bind heparin, and several heparin-binding proteins (HBPs) have been identified in bull seminal plasma. This study had three objectives: 1) to identify production sites of seminal plasma HBPs, 2) to determine which HBPs bound to cauda epididymal sperm, and 3) to determine whether presence of HBPs was testosterone dependent. Proteins from bull or rat seminal vesicles, prostates, and bulbourethral glands were separated by heparin affinity high-performance liquid chromatography. HBPs were found in all accessory glands of rats and bulls, but the major source of bovine seminal plasma HBPs appeared to be seminal vesicles. Between 25% and 50% of the protein from each gland bound to the heparin column, and NaCl concentrations required to elute proteins ranged from 0.15 to 1.4 M. One-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that major HBPs were relatively small, with molecular weights between 13 and 31 kDa, but some HBPs also exhibited higher molecular weights, between 40 and 100 kDa. Radioiodinated HBPs from each bovine gland were incubated with epididymal sperm. Labeled HBPs binding to sperm exhibited molecular weights of 14, 16, 24, and 30 kDa as determined by SDS-PAGE and autoradiography. The HBP content of the accessory sex glands decreased significantly in castrated rats and was restored to levels of sham-operated controls by testosterone replacement. Heparin-binding proteins may play a role in fertilization by attaching to sperm surfaces, enabling heparin-like glycosaminoglycans in the female reproductive tract to induce capacitation.
肝素可与牛精子结合并在体外刺激精子获能。精浆会改变附睾精子与肝素结合的能力,并且已在公牛精浆中鉴定出几种肝素结合蛋白(HBP)。本研究有三个目标:1)确定精浆HBP的产生部位;2)确定哪些HBP与附睾尾精子结合;3)确定HBP的存在是否依赖于睾酮。通过肝素亲和高效液相色谱法分离来自公牛或大鼠精囊、前列腺和尿道球腺的蛋白质。在大鼠和公牛的所有附属腺体中均发现了HBP,但公牛精浆HBP的主要来源似乎是精囊。每个腺体中25%至50%的蛋白质与肝素柱结合,洗脱蛋白质所需的NaCl浓度范围为0.15至1.4M。一维十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)显示,主要的HBP相对较小,分子量在13至31kDa之间,但一些HBP也表现出较高的分子量,在40至100kDa之间。将来自每个牛腺体的放射性碘化HBP与附睾精子一起孵育。通过SDS-PAGE和放射自显影测定,与精子结合的标记HBP的分子量为14、16、24和30kDa。去势大鼠附属性腺的HBP含量显著降低,通过睾酮替代可恢复到假手术对照组的水平。肝素结合蛋白可能通过附着于精子表面在受精过程中发挥作用,使雌性生殖道中的类肝素糖胺聚糖能够诱导精子获能。