Validation of RNA isolated from milk fat globules to profile mammary epithelial cell expression during lactation and transcriptional response to a bacterial infection
Validation of RNA isolated from milk fat globules to profile mammary epithelial cell expression during lactation and transcriptional response to a bacterial infection
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DOI：
10.3168/jds.2012-5604
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发表时间：
2012-10-01
影响因子：
3.5
通讯作者：
Martin, P.
中科院分区：
文献类型：
--
作者：
Brenaut, P.;Bangera, R.;Martin, P.
乳腺炎是一种乳腺炎症，是世界范围内奶牛反刍动物最昂贵的传染病。虽然它受到相当大的关注，但宿主反应的早期步骤仍然很不明确。在这里，我们报告了一种非侵入性的方法，使用乳脂球（MFG）作为乳腺RNA的来源，以遵循细菌感染过程中乳腺上皮细胞（MEC）的全球转录反应的动态。我们首先评估从MFG中分离的RNA是MEG RNA的代表;然后评估MFG RNA是否可用于监测MEG对感染的反应。从山羊MFG获得足够高产量的优质RNA（RNA完整性数值范围在6.7和8.7之间）用于随后的分析。评估了巨噬细胞和中性粒细胞对MFG的污染（这些细胞可能在奶油分层过程中被捕获），当使用细胞类型特异性标记物的定量实时PCR时，结果显示污染足够弱（
Mastitis, an inflammation of the mammary gland, is the most costly infectious disease of dairy ruminants worldwide. Although it receives considerable attention, the early steps of the host response remain poorly defined. Here, we report a noninvasive method using milk fat globules (MFG) as a source of mammary RNA to follow the dynamics of the global transcriptional response of mammary epithelial cells (MEC) during the course of a bacterial infection. We first assessed that RNA isolated from MFG were representative of MEG RNA; we then evaluated whether MFG RNA could be used to monitor the MEG response to infection. Sufficiently high yields of good-quality RNA (RNA integrity numbers ranging between 6.7 and 8.7) were obtained from goat MFG for subsequent analyses. Contamination of MFG by macrophages and neutrophils, which can be trapped during creaming, was assessed and when using quantitative real-time PCR for cell-type specific markers, was shown to be weak enough (