Bodu Mustafa, Hitit Mustafa, Donmez Huseyin, Kaya Abdullah, Ugur Muhammet Rasit, Memili Erdoğan
College of Agriculture, Food and Natural Resources, Cooperative Agricultural Research Center, Prairie View A&M University, Prairie View, TX 77445, USA.
Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Selcuk University, Konya 42005, Türkiye.
Int J Mol Sci. 2025 Mar 17;26(6):2690. doi: 10.3390/ijms26062690.
The identification of molecular markers for fertility is critical for the sustainability of livestock production. We profiled small non-coding RNAs (sncRNAs) in sperm from rams with high fertility (HF) and low fertility (LF) phenotypes to uncover their roles in ram sperm fertility. Rams were categorized into high-fertility (HF, = 31; 94.5 ± 2.8%) and low-fertility (LF, = 25; 83.1 ± 5.73%) phenotypes based on pregnancy rates (average 89.4 ± 7.2%). From these, sperm samples of HF ( = 4; pregnancy rate 99.2 ± 1.6%) and LF ( = 4; pregnancy rate 73.6 ± 4.4%) rams underwent sncRNA sequencing. Small RNA sequencing produced 14,962,876 reads in LF rams and 17,401,094 reads in HF rams, showing distinct sncRNA biotypes, including miRNAs, tRNAs, snoRNAs, snRNAs, and rRNAs. Among these, miRNAs comprised 7.12% of reads in LF rams and 3.78% in HF rams, while rRNAs and repeats formed significant proportions in both groups. A total of 1673 known and 627 novel miRNAs were identified, with 227 differentially expressed miRNAs between the HF and LF groups. We showed that key miRNAs, such as oar-miR-200b and oar-miR-370-3p, were upregulated in HF sperm, while downregulated miRNAs in LF, such as oar-miR-26b and oar-let-7d, were associated with impaired sperm function and DNA fragmentation. A functional enrichment analysis of miRNA target genes highlighted pathways related to ribonucleoprotein complex biogenesis, RNA processing, and gene expression regulation. These findings establish the critical role of sperm sncRNAs as regulators of fertility and potential biomarkers in breeding soundness tests for the precision farming of livestock for global food security.
鉴定生育力的分子标记对于畜牧生产的可持续性至关重要。我们对具有高生育力（HF）和低生育力（LF）表型的公羊精子中的小非编码RNA（sncRNA）进行了分析，以揭示它们在公羊精子生育力中的作用。根据妊娠率（平均89.4±7.2%），公羊被分为高生育力（HF，n = 31；94.5±2.8%）和低生育力（LF，n = 25；83.1±5.73%）表型。从中选取了HF（n = 4；妊娠率99.2±1.6%）和LF（n = 4；妊娠率73.6±4.4%）公羊的精子样本进行sncRNA测序。小RNA测序在LF公羊中产生了14,962,876条读数，在HF公羊中产生了17,401,094条读数，显示出不同的sncRNA生物类型，包括miRNA、tRNA、snoRNA、snRNA和rRNA。其中，miRNA在LF公羊的读数中占7.12%，在HF公羊中占3.78%，而rRNA和重复序列在两组中都占很大比例。共鉴定出1673个已知miRNA和627个新miRNA，HF组和LF组之间有227个差异表达的miRNA。我们发现，关键miRNA，如oar-miR-200b和oar-miR-370-3p，在HF精子中上调，而LF中下调的miRNA，如oar-miR-26b和oar-let-7d，与精子功能受损和DNA片段化有关。对miRNA靶基因的功能富集分析突出了与核糖核蛋白复合体生物发生、RNA加工和基因表达调控相关的途径。这些发现确立了精子sncRNA作为生育力调节剂的关键作用，以及在畜牧精准养殖以保障全球粮食安全的繁殖健全性测试中的潜在生物标志物作用。
