Shandong Research Centre for Stem Cell Engineering, Shandong Province, PR China.
Mol Cell Proteomics. 2010 Nov;9(11):2517-28. doi: 10.1074/mcp.M110.001719. Epub 2010 Aug 24.
The mammalian spermatozoon has many cellular compartments, such as head and tail, permitting it to interact with the female reproductive tract and fertilize the egg. It acquires this fertilizing potential during transit through the epididymis, which secretes proteins that coat different sperm domains. Optimal levels of these proteins provide the spermatozoon with its ability to move to, bind to, fuse with, and penetrate the egg; otherwise male infertility results. As few human epididymal proteins have been characterized, this work was performed to generate a database of human epididymal sperm-located proteins involved in maturation. Two-dimensional gel electrophoresis of epididymal tissue and luminal fluid proteins, followed by identification using MALDI-TOF/MS or MALDI-TOF/TOF, revealed over a thousand spots in gels comprising 745 abundant nonstructural proteins, 408 in luminal fluids, of which 207 were present on spermatozoa. Antibodies raised to 619 recombinant or synthetic peptides, used in Western blots, histological sections, and washed sperm preparations to confirm antibody quality and protein expression, indicated their regional location in the epididymal epithelium and highly specific locations on washed functional spermatozoa. Sperm function tests suggested the role of some proteins in motility and protection against oxidative attack. A large database of these proteins, characterized by size, pI, chromosomal location, and function, was given a unified terminology reflecting their sperm domain location. These novel, secreted human epididymal proteins are potential targets for a posttesticular contraceptive acting to provide rapid, reversible, functional sterility in men and they are also biomarkers that could be used in noninvasive assessments of male fertility.
哺乳动物精子具有许多细胞区室，如头部和尾部，使其能够与雌性生殖道相互作用并使卵子受精。它在通过附睾的过程中获得这种受精能力，附睾分泌的蛋白质覆盖不同的精子区域。这些蛋白质的最佳水平赋予精子向卵子移动、与卵子结合、融合和穿透卵子的能力；否则会导致男性不育。由于很少有人类附睾蛋白被表征，因此进行了这项工作以生成参与成熟的人类附睾精子定位蛋白的数据库。对附睾组织和管腔液蛋白进行二维凝胶电泳，然后使用 MALDI-TOF/MS 或 MALDI-TOF/TOF 进行鉴定，在包含 745 种丰富非结构蛋白的凝胶中显示出超过一千个斑点，管腔液中存在 408 种，其中 207 种存在于精子上。针对 619 种重组或合成肽产生的抗体，用于 Western blot、组织学切片和洗涤精子制剂，以确认抗体质量和蛋白表达，表明它们在附睾上皮中的区域定位以及在洗涤后的功能性精子上的高度特异性定位。精子功能测试表明，一些蛋白质在运动和防止氧化攻击方面发挥作用。这些蛋白质的大型数据库，其特征是大小、pI、染色体位置和功能，被赋予了统一的术语，反映了它们在精子区室中的位置。这些新的、分泌的人类附睾蛋白是一种潜在的睾丸后避孕靶点，可用于提供男性快速、可逆、功能性不育，并且它们也是可用于非侵入性评估男性生育力的生物标志物。
