Laboratory of Diagnostics for Reproductive Medicine, Institute of Biotechnology, Academy of Sciences of the Czech Republic, v.v.i., Vídenská 1083, 142 20, Prague, Czech Republic.
Cell Tissue Res. 2009 Nov;338(2):303-11. doi: 10.1007/s00441-009-0876-9. Epub 2009 Oct 8.
Proteinases and proteinase inhibitors play key roles in almost all physiological processes. Proteinase inhibitors are present in all tissues and body fluids. They interfere with the activity of proteinases and thus maintain homeostasis. The main role of proteinase inhibitors in the reproductive tract is the inactivation of prematurely released hydrolytic enzymes from damaged spermatozoa and the protection of reproductive tracts and spermatozoa against proteolytic degradation. In the boar reproductive system, acrosin inhibitors are found in seminal plasma and on spermatozoa. The amino acid sequence of seminal plasma and sperm-associated acrosin inhibitors is 90% identical, and their biochemical properties have been completely resolved. However, their origin and localization have not been fully elucidated. Using rabbit polyclonal antibody, we have studied the expression and localization of the seminal plasma acrosin inhibitor in the boar reproductive tract. The antibody recognizes a 12-kDa band in extracts from the cauda epididymidis, seminal vesicles, prostate, and Cowper's glands, and immunofluorescence has revealed the acrosin inhibitor in the epithelium and lumen of these organs. Gene expression of the acrosin inhibitor has been studied by reverse transcription together with the polymerase chain reaction. Acrosin inhibitor mRNA transcript is detectable in the epididymis, seminal vesicles, prostate, and Cowper's glands. The antibody has localized the acrosin inhibitor on the surface of epididymal and ejaculated spermatozoa in the acrosomal region. In extracts from epididymal and ejaculated spermatozoa, the specific antibody recognizes acrosin inhibitor at 8 kDa and 12 kDa. The presence of acrosin inhibitor on the sperm surface as a protective molecule for receptors mediating the sperm-zona pellucida binding suggests that it is crucial for the reproductive process.
蛋白酶和蛋白酶抑制剂在几乎所有生理过程中都发挥着关键作用。蛋白酶抑制剂存在于所有组织和体液中。它们干扰蛋白酶的活性，从而维持体内平衡。蛋白酶抑制剂在生殖道中的主要作用是使从受损精子中过早释放的水解酶失活，并保护生殖道和精子免受蛋白水解降解。在猪的生殖系统中，顶体酶抑制剂存在于精液中和精子上。精液和与精子相关的顶体酶抑制剂的氨基酸序列有 90%相同，其生化特性已完全解析。然而，它们的起源和定位尚未完全阐明。我们使用兔多克隆抗体研究了猪生殖系统中精液顶体酶抑制剂的表达和定位。该抗体在附睾尾部、精囊、前列腺和库珀氏腺的提取物中识别出 12 kDa 的条带，免疫荧光显示顶体酶抑制剂存在于这些器官的上皮和管腔中。通过逆转录聚合酶链反应研究了顶体酶抑制剂的基因表达。在附睾、精囊、前列腺和库珀氏腺中均可检测到顶体酶抑制剂的 mRNA 转录本。该抗体将顶体酶抑制剂定位于附睾和射出精子的顶体区域表面。在附睾和射出精子的提取物中，特异性抗体在 8 kDa 和 12 kDa 处识别顶体酶抑制剂。顶体酶抑制剂作为介导精子与透明带结合的受体的保护分子存在于精子表面，这表明它对生殖过程至关重要。
