Department of Animal Sciences, Endocrinology, and Animal Biosciences Program, Rutgers University, New Brunswick, New Jersey, USA.
HudsonAlpha Institute for Biotechnology, Huntsville, Alabama, USA.
Biol Reprod. 2017 Feb 1;96(2):327-340. doi: 10.1093/biolre/iox001.
Factors delivered to offspring in colostrum within 2 days of birth support neonatal porcine uterine development. The uterine mRNA transcriptome is affected by age and nursing during this period. Whether uterine microRNA (miRNA) expression is affected similarly is unknown. Objectives were to (1) determine effects of age and nursing on porcine uterine miRNA expression between birth and postnatal day (PND) 2 using miRNA sequencing (miRNAseq) and; (2) define affected miRNA–mRNA interactions and associated biological processes using integrated target prediction analysis. At birth (PND 0), gilts were euthanized, nursed ad libitum, or gavage-fed milk replacer for 48 h. Uteri were collected at birth or 50 h postnatal. MicroRNAseq data were validated using quantitative real-time PCR. Targets were predicted using an established mRNA database generated from the same tissues. For PND 2 versus PND 0 comparisons, 31 differentially expressed (DE) miRNAs were identified for nursed, and 42 DE miRNAs were identified for replacer-fed gilts. Six DE miRNAs were identified for nursed versus replacer-fed gilts on PND 2. Target prediction for inversely correlated DE miRNA–mRNA pairings indicated 20 miRNAs targeting 251 mRNAs in nursed, versus 29 miRNAs targeting 585 mRNAs in replacer-fed gilts for PND 2 versus PND 0 comparisons, and 5 miRNAs targeting 81 mRNAs for nursed versus replacer-fed gilts on PND 2. Biological processes predicted to be affected by age and nursing included cell-to-cell signaling, cell morphology, and tissue morphology. Results indicate novel age- and lactocrine-sensitive miRNA–mRNA relationships associated with porcine neonatal uterine development between birth and PND 2
产后 2 天内初乳传递给后代的因子支持新生猪子宫发育。在此期间,子宫 mRNA 转录组受年龄和哺乳的影响。子宫 microRNA(miRNA)表达是否受到类似影响尚不清楚。目的是(1)使用 miRNA 测序(miRNAseq)确定出生后和产后第 2 天(PND2)年龄和哺乳对猪子宫 miRNA 表达的影响;(2)使用综合靶标预测分析确定受影响的 miRNA-mRNA 相互作用和相关的生物学过程。在出生时(PND0),母猪被安乐死,自由哺乳,或灌喂代乳 48 小时。产后 0 或 50 小时收集子宫。使用定量实时 PCR 验证 miRNAseq 数据。使用从相同组织生成的成熟 mRNA 数据库预测靶标。对于 PND2 与 PND0 比较,自由哺乳的猪有 31 个差异表达(DE)miRNA,代乳喂养的猪有 42 个 DE miRNA。在 PND2 时,自由哺乳与代乳喂养的猪有 6 个 DE miRNA。反向相关的 DE miRNA-mRNA 配对靶预测表明,在 PND2 时,自由哺乳的猪有 20 个 miRNA 靶向 251 个 mRNA,代乳喂养的猪有 29 个 miRNA 靶向 585 个 mRNA,而自由哺乳与代乳喂养的猪有 5 个 miRNA 靶向 81 个 mRNA。预测受年龄和哺乳影响的生物学过程包括细胞间信号转导、细胞形态和组织形态。结果表明,在出生后至 PND2 期间,与新生猪子宫发育相关的新型年龄和乳泌素敏感的 miRNA-mRNA 关系。