Sakumoto Ryosuke, Hayashi Ken-Go, Hosoe Misa, Iga Kosuke, Kizaki Keiichiro, Okuda Kiyoshi
Animal Physiology Research Unit, National Institute of Agrobiological Sciences, Ibaraki 305-0901, Japan.
J Reprod Dev. 2015;61(1):42-8. doi: 10.1262/jrd.2014-101. Epub 2014 Nov 9.
To determine functional differences between the corpus luteum (CL) of the estrous cycle and pregnancy in cows, gene expression profiles were compared using a 15 K bovine oligo DNA microarray. In the pregnant CL at days 20-25, 40-45 and 150-160, the expressions of 138, 265 and 455 genes differed by a factor of > 2-fold (P < 0.05) from their expressions in the cyclic CL (days 10-12 of the estrous cycle). Messenger RNA expressions of chemokines (eotaxin, lymphotactin and ENA-78) and their receptors (CCR3, XCR1 and CXCR2) were validated by quantitative real-time PCR. Transcripts of eotaxin were more abundant in the CL at days 40-45 and 150-160 of pregnancy than in the cyclic CL (P < 0.01). In contrast, the mRNA expressions of lymphotactin, ENA-78 and XCR1 were lower in the CL of pregnancy (P < 0.05). Messenger RNAs of CCR3 and CXCR2 were similarly detected both in the cyclic and pregnant CL. Tissue protein levels of eotaxin were significantly higher in the CL at days 150-160 of pregnancy than in the CL at other stages, whereas the lymphotactin protein levels in the CL at days 20-25 of pregnancy were lower (P < 0.05). Immunohistochemical staining showed that CCR3 was expressed in the luteal cells and that XCR1 was expressed in both the luteal cells and endothelial cells. Collectively, the different gene expression profiles may contribute to functional differences between the cyclic and pregnant CL, and chemokines including eotaxin and lymphotactin may regulate CL function during pregnancy in cows.
为了确定奶牛发情周期黄体(CL)与妊娠黄体之间的功能差异,使用15K牛寡核苷酸DNA微阵列比较了基因表达谱。在妊娠第20 - 25天、40 - 45天和150 - 160天的妊娠黄体中,分别有138、265和455个基因的表达与发情周期黄体(发情周期第10 - 12天)相比差异超过2倍(P < 0.05)。通过定量实时PCR验证了趋化因子(嗜酸性粒细胞趋化因子、淋巴细胞趋化因子和ENA - 78)及其受体(CCR3、XCR1和CXCR2)的信使RNA表达。嗜酸性粒细胞趋化因子的转录本在妊娠第40 - 45天和150 - 160天的黄体中比在发情周期黄体中更丰富(P < 0.01)。相反,淋巴细胞趋化因子、ENA - 78和XCR1的mRNA表达在妊娠黄体中较低(P < 0.05)。CCR3和CXCR2的信使RNA在发情周期黄体和妊娠黄体中均有类似表达。妊娠第150 - 160天黄体中嗜酸性粒细胞趋化因子的组织蛋白水平显著高于其他阶段的黄体,而妊娠第20 - 25天黄体中淋巴细胞趋化因子蛋白水平较低(P < 0.05)。免疫组织化学染色显示CCR3在黄体细胞中表达,XCR1在黄体细胞和内皮细胞中均有表达。总体而言,不同的基因表达谱可能导致发情周期黄体与妊娠黄体之间的功能差异,包括嗜酸性粒细胞趋化因子和淋巴细胞趋化因子在内的趋化因子可能调节奶牛妊娠期间的黄体功能。