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首页> 外文期刊>Journal of Reproductive Immunology >Identification and isolation of boar sperm specific antigens with potential role in sperm-egg interaction.
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Identification and isolation of boar sperm specific antigens with potential role in sperm-egg interaction.

机译：鉴定和分离在精卵相互作用中具有潜在作用的公猪精子特异性抗原。

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Investigations on specific and functionally active sperm antigens would bring about the elucidation of the mechanisms of gamete interaction and help the search to new approaches for prognosis and regulation of fertility. Previously, we have produced a polyclonal rabbit anti-boar spermatozoa antibody (RABSA) that might affect the fertilizing capacity of boar spermatozoa. The sperm specificity of RABSA was demonstrated by double immunodiffusion, immunoelectrophoresis and ELISA against boar spermatozoa, as well as against saline extracts of boar reproductive and somatic organs. Using indirect immunofluorescence (IIF) test, here we provide evidence that RABSA stained the acrosomes of ejaculated and capacitated boar and human spermatozoa, the fluorescence being intensified on the equatorial region after the acrosome reaction. The RABSA cognate antigen/s is a subject of interest because of their specific localization in sperm structures, which is shown to be a binding and/or fusion competence region. Using ion-exchange (Heparin-Sepharose) chromatography, we eluted an antigen with molecular mass 60 kDa (Ag60) in SDS-PAGE from NP40 extracts of capacitated boar spermatozoa. In Western blot, RABSA recognized specifically this antigen. The Ag60 did not affect the sperm-ligand activity of zona pellucida in a porcine sperm-zona binding assay. IIF experiments showed that zona-free porcine oocytes preincubated with Ag60 and RABSA presented fluorescent labeling over the entire egg surface. The biological and IIF experiments provide evidence supporting the involvement of Ag60 in functional steps required for sperm-egg binding and/or fusion, but not sperm-zona pellucida binding.
机译：对特异性和功能活跃的精子抗原的研究将阐明配子相互作用的机制，并有助于寻找预后和调节生育能力的新方法。以前，我们已经生产了可能影响公猪精子受精能力的多克隆兔抗公猪精子抗体（RABSA）。 RABSA的精子特异性通过针对公猪精子以及公猪生殖器官和体细胞生理盐水的双重免疫扩散，免疫电泳和ELISA证实。使用间接免疫荧光（IIF）测试，在这里我们提供了RABSA染色射精和带电容的公猪和人精子的顶体的荧光，顶体反应后赤道区域的荧光增强。由于其在精子结构中的特异性定位，RABSA关联抗原是感兴趣的主题，已显示其是结合和/或融合能力区域。使用离子交换（肝素-琼脂糖）色谱法，我们从获能力的公猪精子的NP40提取物中在SDS-PAGE中洗脱了分子量为60 kDa（Ag60）的抗原。在蛋白质印迹中，RABSA特异性识别该抗原。在猪的精子-zona结合试验中，Ag60不会影响透明带的精子-配体活性。 IIF实验表明，预先与Ag60和RABSA一起孵育的无透明带状卵母细胞在整个蛋表面上都具有荧光标记。生物学和IIF实验提供的证据支持Ag60参与精卵结合和/或融合所需的功能步骤，但不参与精子-透明带结合。

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