Novel and differentially abundant microRNAs in sperm cells, seminal plasma, and serum of boars due to porcine reproduction and respiratory syndrome virus infection
Novel and differentially abundant microRNAs in sperm cells, seminal plasma, and serum of boars due to porcine reproduction and respiratory syndrome virus infection
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DOI：
10.1016/j.anireprosci.2018.10.007
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发表时间：
2018-12-01
影响因子：
2.2
通讯作者：
Pratt, Scott L.
中科院分区：
文献类型：
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作者：
Calcatera, Samantha M.;Reicks, Darwin;Pratt, Scott L.
本研究的目的是鉴定和确定病毒感染前后公猪精子、精浆 (SP) 和血清中 miRNA 的相对丰度。对感兴趣的 miRNA 的预测靶标进行功能富集分析。公猪 (n = 6) 接种猪繁殖与呼吸综合征病毒 (PRRSv) 1.8-4 株（第 0 天）。在第-2天和第6天收集精液和血清。分离精子和SP，将等分试样快速冷冻并储存在-80℃。将血清冷冻并储存在-80℃。从精子和SP样品中分离总RNA并进行RNA测序。使用来自血清、精子和SP的第-2天和第6天的RNA样品进行微阵列分析。使用 miRanda 3.3a 预测潜在的 miRNA 靶标，然后分析靶标的基因本体 (Gene Ontology) 和 InterPro 术语的富集，如果使用 Bonferroni 校正 P < 0.01，则认为是富集。当比较第-2天和第6天时，微阵列分析得出在精子、血清和SP中丰度分别存在差异的83、13和10个miRNA。富集分析的结果表明，与猪基因组相比，在精子、SP和血清中丰度分别存在差异的35、9和5个miRNA的预测靶标具有富集的功能和/或保守蛋白结构域。为精子、SP 和血清确定了 P2X 嘌呤受体的丰富术语。鉴定了精子和血清转录物的细胞粘附的丰富术语。为精子和 SP 转录本鉴定了丰富的细胞信号传导术语。
The objectives of this study were to identify and determine relative abundance of miRNAs in boar sperm, seminal plasma (SP), and serum pre- and post-viral infection. Functional enrichment analyses on predicted targets of miRNAs of interest were performed. Boars (n = 6) were inoculated with porcine reproductive and respiratory syndrome virus (PRRSv) strain 1.8-4 (Day 0). Semen and serum were collected on Day -2 and 6. Sperm and SP were separated and aliquots were flash frozen and stored at -80 degrees C. Serum was frozen and stored at -80 degrees C. Total RNA was isolated from sperm and SP samples and subjected to RNA sequencing. Microarray analysis was performed using the Day -2 and 6 RNA samples from serum, sperm and SP. Potential miRNA targets were predicted using miRanda 3.3a and targets were then analyzed for enrichment of Gene Ontology) and InterPro terms and were considered to be enriched if P < 0.01 using the Bonferroni correction. Microarray analyses resulted in 83, 13, and 10 miRNAs with differences in abundances in sperm, serum, and SP, respectively, when comparing Day -2 and 6. Results from enrichment analyses indicated that the predicted targets of 35, nine, and five miRNAs with differences in abundances for sperm, SP, and serum, respectively, that have functions and/or conserved protein domains that are enriched when compared to the pig genome. Enriched terms for P2X purinoceptors were identified for sperm, SP and serum. Enriched terms for cell adhesion were identified for sperm and serum transcripts. Enriched terms for cell signaling were identified for sperm and SP transcripts.