Binelli Mario, Scolari Saara C, Pugliesi Guilherme, Van Hoeck Veerle, Gonella-Diaza Angela M, Andrade Sónia C S, Gasparin Gustavo R, Coutinho Luiz L
Laboratório de Fisiologia e Endocrinologia Molecular, Department of Animal Reproduction, FMVZ-USP, Pirassununga, Sao Paulo, Brazil.
Laboratório de Genética Animal, Departamento de Zootecnia, ESALQ-USP, Pirassununga, Sao Paulo, Brazil.
PLoS One. 2015 Apr 7;10(4):e0122874. doi: 10.1371/journal.pone.0122874. eCollection 2015.
Pregnancy success is critical to the profitability of cattle operations. However, the molecular events driving the uterine tissue towards embryo receptivity are poorly understood. This study aimed to characterize the uterine transcriptome profiles of pregnant (P) versus non-pregnant (NP) cows during early pregnancy and attempted to define a potential set of marker genes that can be valuable for predicting pregnancy outcome. Therefore, beef cows were synchronized (n=51) and artificially inseminated (n=36) at detected estrus. Six days after AI (D6), jugular blood samples and a biopsy from the uterine horn contralateral to the ovary containing the corpus luteum were collected. Based on pregnancy outcome on D30, samples were retrospectively allocated to the following groups: P (n=6) and NP (n=5). Both groups had similar plasma progesterone concentrations on D6. Uterine biopsies were submitted to RNA-Seq analysis in a Illumina platform. The 272,685,768 million filtered reads were mapped to the Bos Taurus reference genome and 14,654 genes were analyzed for differential expression between groups. Transcriptome data showed that 216 genes are differently expressed when comparing NP versus P uterine tissue (Padj ≤ 0.1). More specifically, 36 genes were up-regulated in P cows and 180 are up-regulated in NP cows. Functional enrichment and pathway analyses revealed enriched expression of genes associated with extracellular matrix remodeling in the NP cows and nucleotide binding, microsome and vesicular fraction in the P cows. From the 40 top-ranked genes, the transcript levels of nine genes were re-evaluated using qRT-PCR. In conclusion, this study characterized a unique set of genes, expressed in the uterus 6 days after insemination, that indicate a receptive state leading to pregnancy success. Furthermore, expression of such genes can be used as potential markers to efficiently predict pregnancy success.
妊娠成功对于养牛业的盈利能力至关重要。然而,驱动子宫组织达到胚胎接受性的分子事件却知之甚少。本研究旨在描绘妊娠早期怀孕(P)与未怀孕(NP)母牛的子宫转录组图谱,并试图确定一组可能对预测妊娠结局有价值的标记基因。因此,对肉牛母牛进行同期发情处理(n = 51),并在检测到发情时进行人工授精(n = 36)。人工授精后6天(D6),采集颈静脉血样以及来自与含有黄体的卵巢对侧子宫角的活检组织。根据第30天的妊娠结局,将样本回顾性地分为以下几组:P组(n = 6)和NP组(n = 5)。两组在D6时血浆孕酮浓度相似。子宫活检组织在Illumina平台上进行RNA测序分析。272685768百万条过滤后的 reads 被映射到牛参考基因组,并分析了14654个基因在两组间的差异表达。转录组数据显示,比较NP组与P组子宫组织时,有216个基因表达不同(Padj≤0.1)。更具体地说,36个基因在P组母牛中上调,180个基因在NP组母牛中上调。功能富集和通路分析显示,NP组母牛中与细胞外基质重塑相关的基因表达富集,而P组母牛中与核苷酸结合、微粒体和囊泡部分相关的基因表达富集。从排名前40的基因中,使用qRT-PCR重新评估了9个基因的转录水平。总之,本研究描绘了一组独特的基因,这些基因在授精后6天在子宫中表达,表明处于导致妊娠成功的接受状态。此外,这些基因的表达可作为潜在标记物,用于有效预测妊娠成功。