Mahé Coline, Mowinska Aleksandra Maria, Albrecht Elke, Reynaud Karine, Lavigne Régis, Com Emmanuelle, Pineau Charles, Mermillod Pascal, Saint-Dizier Marie, Schoen Jennifer
Reproduction. 2025 Jul 17;170(2). doi: 10.1530/REP-24-0390. Print 2025 Aug 1.
The molecular interactions between spermatozoa and the oviduct epithelium that are involved in capacitation and sperm availability during fertilization still remain largely unknown in many species. This study provides comprehensive proteomes of the apical and basal part of the bovine oviduct epithelium (isthmus and ampulla, pre- and post-ovulatory) and presents new protein candidates for sperm binding in the bovine functional sperm reservoir.
The proximal region of the oviduct (isthmus) serves as a sperm reservoir in many mammalian species. This reservoir is composed of epithelial cells that bind sperm primarily to their apical cilia and ensure the survival of high-quality sperm until ovulation. The species-specific molecular interactions between spermatozoa and the oviduct epithelium, however, are still poorly understood. The aim of this study is to identify new candidates for sperm-interacting proteins in the bovine reservoir using laser capture proteomics. Ipsilateral oviducts were collected from cyclic cows at pre- and post-ovulatory stages. Isthmus and ampulla tissue samples were fixed, paraffin-embedded, sectioned and subjected to laser microdissection to establish pools of microsamples separating the apical part of the epithelium, containing potential sperm-interacting proteins, from the basal part, containing proteins not currently located or secreted on the cell surface. Proteins were analyzed by nanoLC-MS/MS, and differentially abundant proteins (DAPs) were identified using t-tests. A total of 505 and 512 proteins could be identified in apical and basal microsamples, respectively. Only 8/141 regional and 1/79 cycle stage-related DAPs were shared between both epithelium parts, indicating a selective regulation of protein expression. Of the apical proteins, 19 were predicted to be candidates for sperm interactions, including annexins (ANXA) 1, 2, 5 and 8; oviduct glycoprotein 1 (OVGP1); voltage-dependent anion-selective channel proteins (VDAC) 1 and 2; apolipoprotein A1 (APOA1). This work provides the first proteomic characterization of microdissected cellular compartments of the bovine oviduct epithelium and presents new candidates for improving sperm quality in assisted reproductive technologies.
在许多物种中，精子与输卵管上皮之间参与受精过程中获能和精子可用性的分子相互作用在很大程度上仍不清楚。本研究提供了牛输卵管上皮（峡部和壶腹部，排卵前和排卵后）顶端和基部的综合蛋白质组，并提出了牛功能性精子储存库中精子结合的新蛋白质候选物。
在许多哺乳动物物种中，输卵管的近端区域（峡部）充当精子储存库。这个储存库由上皮细胞组成，这些上皮细胞主要将精子结合到其顶端纤毛上，并确保高质量精子在排卵前存活。然而，精子与输卵管上皮之间物种特异性的分子相互作用仍知之甚少。本研究的目的是使用激光捕获蛋白质组学鉴定牛储存库中与精子相互作用蛋白的新候选物。在排卵前和排卵后阶段从周期性奶牛收集同侧输卵管。峡部和壶腹部组织样本进行固定、石蜡包埋、切片并进行激光显微切割，以建立微样本库，将含有潜在精子相互作用蛋白的上皮顶端部分与含有目前不在细胞表面定位或分泌的蛋白质的基部部分分开。通过纳升液相色谱-串联质谱法分析蛋白质，并使用t检验鉴定差异丰富蛋白质（DAP）。在顶端和基部微样本中分别鉴定出总共505和512种蛋白质。上皮两个部分之间仅共享8/141个区域相关和1/79个周期阶段相关的DAP，表明蛋白质表达的选择性调节。在顶端蛋白质中，有19种被预测为精子相互作用的候选物，包括膜联蛋白（ANXA）1、2、5和8；输卵管糖蛋白1（OVGP1）；电压依赖性阴离子选择性通道蛋白（VDAC）1和2；载脂蛋白A1（APOA1）。这项工作提供了牛输卵管上皮显微切割细胞区室的首次蛋白质组学表征，并提出了在辅助生殖技术中改善精子质量的新候选物。
