Solexa sequencing and custom microRNA chip reveal repertoire of microRNAs in mammary gland of bovine suffering from natural infectious mastitis
Solexa sequencing and custom microRNA chip reveal repertoire of microRNAs in mammary gland of bovine suffering from natural infectious mastitis
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Solexa 测序和定制 microRNA 芯片揭示了患有自然传染性乳腺炎的牛乳腺中的 microRNA 库
DOI：
10.1111/age.12628
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发表时间：
2018-02-01
期刊：
影响因子：
2.4
通讯作者：
Huang, Jinming
中科院分区：
文献类型：
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作者：
Ju, Zhihua;Jiang, Qiang;Huang, Jinming
鉴定与先天免疫和炎症反应以及组织损伤相关的microRNA（miRNAs）、靶基因和调控网络对于阐明乳腺炎抗性的分子和遗传机制至关重要。在这项研究中，Solexa测序和定制的miRNA芯片方法的组合被用来分析在自然感染金黄色葡萄球菌（一种广泛的乳腺炎病原体）的晚期阶段牛乳腺中miRNA的表达。我们在健康奶牛和乳腺炎奶牛的乳腺中发现了383个位点，对应于277个已知的和49个推定的新的miRNAs，两个潜在的mitron和266个差异表达的miRNAs。一些相互作用的网络和调节剂参与乳腺炎的易感性，如ALCAM，COL 1A 1，APOP 4，ITIH 4，CRP和纤维蛋白原α（FGA），强调。使用定量实时PCR、原位杂交和双荧光素酶报告基因测定验证了乳腺炎乳腺中靶向FGA的bta-miR-26 a的显著下调和定位。我们认为，在乳腺炎奶牛乳腺中观察到的miRNA变异与感染后期免疫和防御反应的维持、细胞增殖和凋亡以及组织损伤和愈合有关。此外，bta-miR-26 a在乳腺炎中的作用（至少部分通过增强FGA表达介导）涉及宿主防御、炎症和组织损伤。
Identification of microRNAs (miRNAs), target genes and regulatory networks associated with innate immune and inflammatory responses and tissue damage is essential to elucidate the molecular and genetic mechanisms for resistance to mastitis. In this study, a combination of Solexa sequencing and custom miRNA chip approaches was used to profile the expression of miRNAs in bovine mammary gland at the late stage of natural infection with Staphylococcus aureus, a widespread mastitis pathogen. We found 383 loci corresponding to 277 known and 49 putative novel miRNAs, two potential mitrons and 266 differentially expressed miRNAs in the healthy and mastitic cows' mammary glands. Several interaction networks and regulators involved in mastitis susceptibility, such as ALCAM, COL1A1, APOP4, ITIH4, CRP and fibrinogen alpha (FGA), were highlighted. Significant down-regulation and location of bta-miR-26a, which targets FGA in the mastitic mammary glands, were validated using quantitative real-time PCR, in situ hybridization and dual-luciferase reporter assays. We propose that the observed miRNA variations in mammary glands of mastitic cows are related to the maintenance of immune and defense responses, cell proliferation and apoptosis, and tissue injury and healing during the late stage of infection. Furthermore, the effect of bta-miR-26a in mastitis, mediated at least in part by enhancing FGA expression, involves host defense, inflammation and tissue damage.