State Key Laboratory of Animal Biotech Breeding, Key Laboratory of Animal (Poultry) Genetics Breeding and Reproduction of Ministry of Agriculture and Rural Affairs, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100193, China; College of Life Sciences and Food Engineering, Hebei University of Engineering, Handan, 056038, Hebei, China.
State Key Laboratory of Animal Biotech Breeding, Key Laboratory of Animal (Poultry) Genetics Breeding and Reproduction of Ministry of Agriculture and Rural Affairs, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100193, China.
Poult Sci. 2023 Dec;102(12):103099. doi: 10.1016/j.psj.2023.103099. Epub 2023 Sep 12.
The presence of EVs in seminal plasma (SPEVs) suggests their involvement on fertility via transmitting information between the original cells and recipient cells. SPEVs-coupled miRNAs have been shown to affect sperm motility, maturation, and capacitation in mammals, but rarely in poultry species. The present study aims to reveal the profile of SPEVs miRNAs and their potential effect on sperm storage and function in poultry. The SPEVs was successfully isolated from 4 different chicken breeds by ultracentrifugation and verified. Deep sequencing of SPEVs small RNA library of each breed identified 1077 miRNAs in total and 563 shared ones. The top 10 abundant miRNAs (such as miR-10-5p, miR-100-5p, and miR-10a-5p etc.) accounted for around 60% of total SPEVs miRNA reads and are highly conserved across species, predisposing their functional significance. Target genes prediction and functional enrichment analysis indicated that the most abundantly expressed miRNAs may regulate pathways like ubiquitin-mediated proteolysis, endocytosis, mitophagy, glycosphingolipid biosynthesis, fatty acid metabolism, and fatty acid elongation. The high abundant SPEVs-coupled miRNAs were found to target 107 and 64 functionally important mRNAs in the potential recipient cells, sperm and sperm storage tubules (SST) cells, respectively. The pathways that enriched by target mRNAs revealed that the SPEVs-coupled miRNA may rule the fertility by affecting the sperm maturation and regulating the female's immune response and lipid metabolism. In summary, this study presents the distinctive repertoire of SPEVs-coupled miRNAs, and extends our understanding about their potential roles in sperm maturation, capacitation, storage, and fertility, and may help to develop new therapeutic strategies for male infertility and sperm storage.
精子外泌体（SPEVs）的存在表明它们通过在原始细胞和受体细胞之间传递信息参与生育。哺乳动物中已经证实 SPEVs 结合的 miRNA 会影响精子的运动、成熟和获能，但在禽类中很少见。本研究旨在揭示 SPEVs miRNA 的特征及其对禽类精子储存和功能的潜在影响。通过超速离心成功地从 4 个不同的鸡品种中分离出 SPEVs 并进行了验证。对每个品种的 SPEVs 小 RNA 文库进行深度测序，共鉴定出 1077 个 miRNA，其中 563 个是共享的。前 10 个丰富的 miRNA（如 miR-10-5p、miR-100-5p 和 miR-10a-5p 等）占总 SPEVs miRNA 读数的约 60%，在物种间高度保守，这表明它们具有功能意义。靶基因预测和功能富集分析表明，表达最丰富的 miRNA 可能调节泛素介导的蛋白水解、内吞作用、线粒体自噬、糖脂生物合成、脂肪酸代谢和脂肪酸延长等途径。高丰度的 SPEVs 结合 miRNA 被发现可以靶向潜在受体细胞精子和精子储存管（SST）细胞中的 107 和 64 个功能重要的 mRNA。通过靶 mRNA 富集的途径表明，SPEVs 结合的 miRNA 可能通过影响精子成熟和调节女性的免疫反应和脂质代谢来控制生育能力。总之，本研究展示了 SPEVs 结合 miRNA 的独特特征谱，扩展了我们对它们在精子成熟、获能、储存和生育能力中的潜在作用的理解，并可能有助于为男性不育和精子储存开发新的治疗策略。
