Circulating miRNA signatures of early pregnancy in cattle.
Circulating miRNA signatures of early pregnancy in cattle.
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DOI：
10.1186/s12864-016-2529-1
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发表时间：
2016-03-03
期刊：
影响因子：
4.4
通讯作者：
Donadeu FX
中科院分区：
文献类型：
--
作者：
Ioannidis J;Donadeu FX
低生育率仍然是奶牛生产力低下的主要原因。在这种背景下，人们对开发用于准确早期诊断怀孕的新工具非常感兴趣。MicroRNAs(MiRNAs)是一种短的RNA分子，在健康和疾病过程中都参与调节基因的表达。MiRNAs已被证明可以调节卵巢功能、子宫容受性、胚胎发育和胎盘功能。循环中的miRNAs可以为组织功能和疾病提供有用的生物标记物；重要的是，差异miRNA图谱与人类怀孕和先兆子痫有关。这项研究试图确定循环中的miRNAs作为牛早期怀孕的生物标记物的潜力。我们应用Illumina Small-RNA测序技术分析了发情周期0、8和16天的8头未怀孕小母牛和怀孕16和24天的11头小母牛的血浆样本中的miRNAs。我们总共对46个样本进行了测序，每个样本产生了920万个miRNA读数。在怀孕和非怀孕的任何时间点之间，miRNA读取丰度没有差异(FDR >； 0.1)。作为补充方法，我们分析了与发情周期0、8、16天和怀孕24天对应的样本池(3-4个样本/池)(n = 3池/组)。共有16个miRNA在妊娠和非妊娠动物的血浆中差异表达(fdr <； 0.1)。用相同的血浆样本进行RT-qPCR验证证实，miR-26a在怀孕第16天与未怀孕的小母牛相比差异表达(P = 0.043)，而miR-1249在怀孕第16天的小母牛中有上调的趋势(1.6倍；P = 0.081)。在一组独立的小母牛中的进一步验证证实，在怀孕早期血浆miR-26a水平增加，只有在第24天显著(2.0倍；P = 0.027)。通过全基因组分析，我们已经成功地分析了与牛早期怀孕相关的血浆miRNA群体。我们已经确定miR-26a是一个潜在的早期妊娠的循环生物标志物。本文的在线版本(doi：10.1186/s12864-0162529-1)包含补充材料，授权用户可以使用。
Low fertility remains a leading cause of poor productivity in dairy cattle. In this context, there is significant interest in developing novel tools for accurate early diagnosis of pregnancy. MicroRNAs (miRNAs) are short RNA molecules which are critically involved in regulating gene expression during both health and disease. MiRNAs have been shown to regulate ovarian function, uterine receptivity, embryonic development and placental function. Circulating miRNAs can provide useful biomarkers of tissue function and disease; importantly, differential miRNA profiles have been linked to pregnancy and preeclampsia in humans. This study sought to establish the potential of circulating miRNAs as biomarkers of early pregnancy in cattle. We applied Illumina small-RNA sequencing to profile miRNAs in plasma samples collected from eight non-pregnant heifers on Days 0, 8 and 16 of the oestrous cycle and 11 heifers on Days 16 and 24 of pregnancy. We sequenced a total of 46 samples and generated 9.2 million miRNA reads per sample. There were no differences in miRNA read abundance between any of the pregnant and non-pregnant time-points (FDR > 0.1). As a complementary approach, we analysed sample pools (3–4 samples/pool) corresponding to Days 0, 8 and 16 of the oestrous cycle and Day 24 of pregnancy (n = 3 pools/group) using Qiagen PCR arrays. A total of 16 miRNAs were differentially expressed (FDR < 0.1) in plasma between pregnant and non-pregnant animals. RT-qPCR validation using the same plasma samples confirmed that miR-26a was differentially upregulated on Day 16 pregnant relative to non-pregnant heifers (1.7-fold; P = 0.043), whereas miR-1249 tended to be upregulated in Day 16 pregnant heifers (1.6-fold; P = 0.081). Further validation in an independent group of heifers confirmed an increase in plasma miR-26a levels during early pregnancy, which was significant only on Day 24 (2.0-fold; P = 0.027). Through genome-wide analyses we have successfully profiled plasma miRNA populations associated with early pregnancy in cattle. We have identified miR-26a as a potential circulating biomarker of early pregnancy. The online version of this article (doi:10.1186/s12864-016-2529-1) contains supplementary material, which is available to authorized users.