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利用下一代测序技术对牛生育力模型进行输卵管转录谱分析。

Oviductal transcriptional profiling of a bovine fertility model by next-generation sequencing.

作者信息

Gonella-Diaza Angela Maria, da Silva Andrade Sónia Cristina, Sponchiado Mariana, Pugliesi Guilherme, Mesquita Fernando Silveira, Van Hoeck Veerle, de Francisco Strefezzi Ricardo, Gasparin Gustavo R, Coutinho Luiz L, Binelli Mario

机构信息

Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Pirassununga, São Paulo, Brazil.

Departamento de Genética e Biologia Evolutiva, IB-Universidade de São Paulo, Cidade Universitária, São Paulo, Brazil.

出版信息

Genom Data. 2017 Jun 15;13:27-29. doi: 10.1016/j.gdata.2017.06.004. eCollection 2017 Sep.

Abstract

In cattle, the oviduct plays a fundamental role in the reproductive process. Oviductal functions are controlled by the ovarian sex steroids: estradiol and progesterone. Here, we tested the hypothesis that the exposure to contrasting sex steroid milieus differentially impacts the oviductal transcriptional profile. We manipulated growth of the pre-ovulatory follicle to obtain cows that ovulated a larger (LF group) or a smaller (SF group) follicle. The LF group presented greater proestrus/estrus concentrations of estradiol and metaestrus concentrations of progesterone (Gonella-Diaza et al. 2015 [1], Mesquita et al. 2014 [2]). Also, the LF group was associated with greater fertility in timed-artificial insemination programs (Pugliesi et al. 2016 [3]). Cows were slaughtered on day 4 of the estrous cycle and total RNA was extracted from ampulla and isthmus fragments and analyzed by RNAseq. The resulting reads were mapped to the bovine genome ( UMD 3.1, NCBI). The differential expression analyses revealed that 325 and 367 genes in ampulla and 274 and 316 genes in the isthmus were up-regulated and down-regulated in LF samples, respectively. To validate the RNAseq results, transcript abundance of 23 genes was assessed by qPCR and expression patterns were consistent between the two techniques. A functional enrichment analysis was performed using Database for Annotation, Visualization and Integrated Discovery (DAVID) software. Processes enriched in the LF group included tissue morphology changes (extracellular matrix remodeling), cellular changes (proliferation), and secretion changes (growth factors, ions and metal transporters). An overview of the gene expression data was deposited in the NCBI's Gene Expression Omnibus (GEO) and is accessible through the accession number GSE65681. In conclusion, differences in the peri-ovulatory sex steroid milieu modify the oviductal gene expression profiles. Such differences may be associated with the greater fertility of the LF cows. This dataset is useful for further investigations of the oviductal biology and the impact of sex-steroid on the female reproductive tract.

摘要

在牛中,输卵管在生殖过程中起着至关重要的作用。输卵管的功能受卵巢性类固醇——雌二醇和孕酮的控制。在此,我们检验了这样一个假设:暴露于不同的性类固醇环境会对输卵管的转录谱产生不同影响。我们通过控制排卵前卵泡的生长,获得了排卵卵泡较大(LF组)或较小(SF组)的母牛。LF组在发情前期/发情期的雌二醇浓度以及发情后期的孕酮浓度更高(Gonella - Diaza等人,2015年[1];Mesquita等人,2014年[2])。此外,在定时人工授精程序中,LF组的受孕率更高(Pugliesi等人,2016年[3])。在发情周期的第4天屠宰母牛,从壶腹部和峡部片段中提取总RNA,并通过RNA测序进行分析。所得读数被映射到牛基因组(UMD 3.1,NCBI)。差异表达分析显示,LF样本中壶腹部分别有325个和367个基因上调和下调,峡部分别有274个和316个基因上调和下调。为了验证RNA测序结果,通过定量PCR评估了23个基因的转录本丰度,两种技术的表达模式一致。使用注释、可视化和综合发现数据库(DAVID)软件进行了功能富集分析。LF组中富集的过程包括组织形态变化(细胞外基质重塑)、细胞变化(增殖)和分泌变化(生长因子、离子和金属转运蛋白)。基因表达数据的概述已存入NCBI的基因表达综合数据库(GEO),可通过登录号GSE65681获取。总之,排卵周围性类固醇环境的差异会改变输卵管的基因表达谱。这些差异可能与LF组母牛更高的受孕率有关。该数据集有助于进一步研究输卵管生物学以及性类固醇对雌性生殖道的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9dd2/5480813/0c193b4cb2cb/gr1.jpg

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