Swanson Rebecca M, Arick Mark A, Gonçalves Lucas Melo, Davis Dylan Blake, Lawton Stewart Robert, Fontes Pedro Levy Piza, Lemley Caleb O
Department of Animal and Dairy Sciences, Mississippi State University, Mississippi State, MS, USA39762.
Institute for Genomics, Biocomputing, and Biotechnology, Mississippi State University, Mississippi State, MS, USA39762.
J Anim Sci. 2025 Jan 4;103. doi: 10.1093/jas/skaf267.
Young beef bulls are often fed high-energy diets to maximize postweaning growth; however, overnutrition has been associated with decreased semen quality. Seminal plasma has been shown to prime the uterus for early pregnancy events in mammalian species; thus, alterations in this response caused by overnutrition could be detrimental or beneficial. The present study evaluated the impact of seminal plasma from bulls exposed to overnutrition on the transcriptome of endometrial epithelial cells in vitro. Yearling half-sibling beef bulls (n = 20) were stratified by age, body weight, and body condition score before being randomly assigned to one of two diets: 1) a diet formulated to elicit a target average daily gain (ADG) of 1.81 kg/d (High Gain, n = 10) or 2) a diet formulated to elicit a target ADG of 1.22 kg/d (Moderate Gain, n = 10). Seminal plasma was collected from each treatment group at three timepoints and then pooled and passed through a 22 μm sterile filter. Based on preliminary experiments, cells were randomly assigned to 1 of the 3 treatment groups: no treatment (Control), 1% seminal plasma from moderate gain bulls (Moderate Gain), or 1% seminal plasma from high gain bulls (High Gain) and cultured at 38.5 °C, 95% O2, 5% CO2 for 12 h. Experiments were conducted in duplicate per treatment, 3 times for a total of 6 experimental units per treatment group. Following the treatment period, total RNA was extracted from cells and underwent sequencing on the Illumina platform. Raw reads were mapped to the ARS-UCD1.3 Bos taurus RefSeq transcriptome, then converted to gene-level counts. The quasi-likelihood negative binomial generalized log-linear model (glmOL) from edgeR was used to test for significantly differentially expressed genes (DEG) at Benjamin-Hochberg adjusted (B.H. adj) P ≤ 0.05. There were 7,905 DEG (Padj ≤ 0.05) in bEND cells treated with seminal plasma from High Gain bulls compared with the Control. There were 8,933 (Padj ≤ 0.05) DEG in bEND cells treated with seminal plasma from Moderate Gain bulls compared with Control. There were 28 DEG (Padj ≤ 0.05) in bEND cells treated with seminal plasma from High Gain compared with Moderate Gain bulls. Metabolic pathways were depleted in each of these comparisons. In conclusion, seminal plasma alters the bEND cell transcriptome; however, the plane of nutrition had minimal effects.
年轻的肉用公牛通常被饲喂高能量日粮以最大化断奶后的生长;然而,营养过剩与精液质量下降有关。在哺乳动物中,精浆已被证明能使子宫为早期妊娠事件做好准备;因此,营养过剩引起的这种反应变化可能是有害的或有益的。本研究评估了来自营养过剩公牛的精浆对体外子宫内膜上皮细胞转录组的影响。一岁的半同胞肉用公牛(n = 20)在按年龄、体重和体况评分分层后,被随机分配到两种日粮之一:1)一种配制的日粮,目标平均日增重(ADG)为1.81 kg/d(高增重组,n = 10)或2)一种配制的日粮,目标ADG为1.22 kg/d(中等增重组,n = 10)。在三个时间点从每个处理组收集精浆,然后合并并通过22μm无菌过滤器。根据初步实验,将细胞随机分配到3个处理组中的1个:不处理(对照组)、1%中等增重公牛的精浆(中等增重组)或1%高增重公牛的精浆(高增重组),并在38.5℃、95% O2、5% CO2条件下培养12小时。每个处理重复进行实验,共进行3次,每个处理组共有6个实验单位。处理期结束后,从细胞中提取总RNA,并在Illumina平台上进行测序。将原始读数映射到ARS-UCD1.3牛参考转录组,然后转换为基因水平的计数。使用edgeR中的拟似然负二项广义对数线性模型(glmOL)在本杰明-霍奇伯格校正(B.H. adj)P≤0.05时测试显著差异表达基因(DEG)。与对照组相比,用高增重公牛精浆处理的bEND细胞中有7905个DEG(Padj≤0.05)。与对照组相比,用中等增重公牛精浆处理的bEND细胞中有8933个(Padj≤0.05)DEG。与中等增重公牛精浆处理的bEND细胞相比,用高增重公牛精浆处理的bEND细胞中有28个DEG(Padj≤0.05)。在这些比较中的每一个中,代谢途径都减少了。总之,精浆改变了bEND细胞转录组;然而,营养水平的影响最小。