Acuña O S, Avilés M, López-Úbeda R, Guillén-Martínez A, Soriano-Úbeda C, Torrecillas A, Coy P, Izquierdo-Rico M J
Key Department of Cell Biology and Histology, Faculty of Medicine, University of Murcia, 30100, Murcia, Spain.
Key Department of Physiology, Veterinary Faculty, University of Murcia, 30100, Murcia, Spain.
Reprod Fertil Dev. 2017 Nov;29(12):2387-2399. doi: 10.1071/RD16457.
The oviduct undergoes changes under the influence of steroid hormones during the oestrous cycle. However, the molecular mechanisms underlying oviductal regulation are not fully understood. The aim of the present study was to identify the gene expression profile of the porcine oviduct in different stages of the cycle using microarray technology. A systematic study was performed on animals at four different stage: prepubertal gilts, and sows in the preovulatory, postovulatory and luteal phase of the oestrous cycle. The porcine oviduct expressed a total of 4929 genes. Moreover, significant differences in the expression of several genes were detected as the oestrous cycle progressed. Analysis of the differentially expressed genes indicated that a total of 86, 89 and 15 genes were upregulated in prepubertal gilts, preovulatory and luteal sows respectively compared with levels observed in postovulatory sows. Moreover, 80, 51 and 64 genes were downregulated in prepubertal, preovulatory and luteal animals respectively compared with the postovulatory sows. The concentrations of 10 selected transcripts were quantified by real-time reverse transcription-polymerase chain reaction to validate the cDNA array hybridisation data. Conversely, for some genes, localisation of corresponding protein expression in the oviduct was analysed by immunohistochemistry (i.e. cholecystokinin, glutathione peroxidase 2, mucin 1, phosphatidylethanolamine binding protein 4 and tachykinin 3) and mass spectrometry analysis of oviductal fluid allowed identification of peptides from all five proteins. The results of the present study demonstrate that gene expression in the porcine oviduct is clearly regulated during the oestrous cycle, with some oviductal proteins that could be related to several reproductive processes described here for the first time.
在发情周期中,输卵管会在类固醇激素的影响下发生变化。然而,输卵管调节的分子机制尚未完全明确。本研究的目的是利用微阵列技术确定猪输卵管在发情周期不同阶段的基因表达谱。对处于四个不同阶段的动物进行了系统研究:青春期前的小母猪,以及发情周期中排卵前、排卵后和黄体期的母猪。猪输卵管共表达了4929个基因。此外,随着发情周期的进展,检测到几个基因的表达存在显著差异。对差异表达基因的分析表明,与排卵后母猪相比,青春期前小母猪、排卵前和黄体期母猪中分别有86、89和15个基因上调。此外,与排卵后母猪相比,青春期前、排卵前和黄体期动物中分别有80、51和64个基因下调。通过实时逆转录-聚合酶链反应对10个选定转录本的浓度进行定量,以验证cDNA阵列杂交数据。相反,对于一些基因,通过免疫组织化学分析了输卵管中相应蛋白质表达的定位(即胆囊收缩素、谷胱甘肽过氧化物酶2、粘蛋白1、磷脂酰乙醇胺结合蛋白4和速激肽3),并且对输卵管液的质谱分析鉴定出了所有这五种蛋白质的肽段。本研究结果表明,猪输卵管中的基因表达在发情周期中受到明显调控,一些输卵管蛋白可能与这里首次描述的几个生殖过程有关。