Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology and College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, P.R. China.
Wuhan Institute of Animal Husbandry and Veterinary Science, Wuhan Academy of Agricultural Science & Technology, Wuhan, Hubei 430208, P.R. China.
Poult Sci. 2024 Sep;103(9):103928. doi: 10.1016/j.psj.2024.103928. Epub 2024 Jun 5.
Semen quality is an important indicator that can directly affect fertility. In mammals, miRNAs in seminal plasma extracellular vesicles (SPEVs) and sperms can regulate semen quality. However, relevant regulatory mechanism in duck sperms remains largely unclear. In this study, duck SPEVs were isolated and characterized by transmission electron microscopy (TEM), western blot (WB), and nanoparticle tracking analysis (NTA). To identify the important molecules affecting semen quality, we analysed the miRNA expression in sperms and SPEVs of male ducks in high semen quality group ((DHS, DHSE) and low semen quality group (DLS, DLSE). We identified 94 differentially expressed (DE) miRNAs in the comparison of DHS vs. DLS, and 21 DE miRNAs in DHSE vs. DLSE. Target genes of SPEVs DE miRNAs were enriched in ErbB signaling pathway, glycometabolism, and ECM-receptor interaction pathways (P < 0.05), while the target genes of sperm DE miRNAs were enriched in ribosome (P < 0.05). The miRNA-target-pathway interaction network analyses indicated that 5 DE miRNAs (miR-34c-5p, miR-34b-3p, miR-449a, miR-31-5p, and miR-128-1-5p) targeted the largest number of target genes enriched in MAPK, Wnt and calcium signaling pathways, of which FZD9 and ANAPC11 were involved in multiple biological processes related to sperm functions, indicating their regulatory effects on sperm quality. The comparison of DE miRNAs of SPEVs and sperms found that mir-31-5p and novel-273 could potentially serve as biomarkers for semen quality detection. Our findings enhance the insight into the crucial role of SPEV and sperm miRNAs in regulating semen quality and provide a new perspective for subsequent studies.
精液质量是直接影响生育能力的一个重要指标。在哺乳动物中，精液中细胞外囊泡（SPEV）和精子中的 microRNA（miRNA）可以调节精液质量。然而，鸭精子中相关的调节机制在很大程度上尚不清楚。在本研究中，通过透射电子显微镜（TEM）、Western blot（WB）和纳米颗粒跟踪分析（NTA）分离和表征了鸭 SPEV。为了鉴定影响精液质量的重要分子，我们分析了高精液质量组（DHS、DHSE）和低精液质量组（DLS、DLSE）公鸭精子和 SPEV 中的 miRNA 表达。我们在 DHS 与 DLS 的比较中鉴定出 94 个差异表达（DE）miRNA，在 DHSE 与 DLSE 的比较中鉴定出 21 个 DE miRNA。SPEV 的 DE miRNA 的靶基因富集在 ErbB 信号通路、糖代谢和 ECM-受体相互作用通路（P<0.05），而精子的 DE miRNA 的靶基因富集在核糖体（P<0.05）。miRNA-靶基因-通路相互作用网络分析表明，5 个 DE miRNA（miR-34c-5p、miR-34b-3p、miR-449a、miR-31-5p 和 miR-128-1-5p）靶向了 MAPK、Wnt 和钙信号通路中最大数量的靶基因，其中 FZD9 和 ANAPC11 参与了与精子功能相关的多个生物学过程，表明它们对精子质量的调节作用。SPEV 和精子中 DE miRNA 的比较发现，mir-31-5p 和 novel-273 可能可作为精液质量检测的生物标志物。本研究结果增强了对 SPEV 和精子 miRNA 调节精液质量的关键作用的认识，为后续研究提供了新的视角。
