School of Agriculture and Food Science, University College Dublin, Belfield, Dublin, Ireland.
Physiol Genomics. 2012 Aug 17;44(16):799-810. doi: 10.1152/physiolgenomics.00067.2012. Epub 2012 Jul 3.
The aims of this study were to 1) identify the earliest transcriptional response of the bovine endometrium to the presence of the conceptus (using RNAseq), 2) investigate if these genes are regulated by interferon tau (IFNT) in vivo, and 3) determine if they are predictive of the pregnancy status of postpartum dairy cows. RNAseq identified 459 differentially expressed genes (DEGs) between pregnant and cyclic endometria on day 16. Quantitative real-time PCR analysis of selected genes revealed PARP12, ZNFX1, HERC6, IFI16, RNF213, and DDX58 expression increased in pregnant compared with cyclic endometria on day 16 and were directly upregulated by intrauterine infusion of IFNT in vivo for 2 h (P < 0.05). On day 13 following estrous endometrial expression of nine genes increased [ARHGAP1, MGC127874, LIMS2, TBC1D1, FBXL7, C25H16orf71, LOC507810, ZSWIM4, and one novel gene (ENSBTAT00000050193)] and seven genes decreased (SERBP1, SRGAP2, AL7A1, TBK1, F2RL2, MGC128929, and WBSCR17; P < 0.05) in pregnant compared with cyclic heifers. Of these DEGs, significant differences in expression between pregnant and cyclic endometria were maintained on day 16 for F2RL2, LIMS2, LOC507810, MGC127874, TBC1D1, WBSCR17, and ZSWIM4 (P < 0.05) both their expression was not directly regulated by IFNT in vivo. Analysis of the expression of selected interferon-stimulated genes in blood samples from postpartum dairy cows revealed a significant increase (P < 0.05) in expression of ZXFX1, PARP12, SAMD9, and HERC6 on day 18 following artificial insemination in cows subsequently confirmed pregnant compared with cyclic controls. In conclusion, RNAseq identified a number of novel pregnancy-associated genes in the endometrium of cattle during early pregnancy that are not regulated by IFNT in vivo. In addition, a number of genes that are directly regulated by short term exposure to IFNT in vivo are differentially expressed on day 18 following estrus detection in the blood of postpartum dairy cows depending on their pregnancy status.
1）鉴定牛子宫内膜对胚胎存在的最早转录反应（使用 RNAseq），2）研究这些基因是否在体内受干扰素 tau（IFNT）调节，3）确定它们是否可预测产后奶牛的妊娠状态。RNAseq 在妊娠和循环子宫内膜之间鉴定出 16 天的 459 个差异表达基因（DEGs）。对选定基因的定量实时 PCR 分析显示，PARP12、ZNFX1、HERC6、IFI16、RNF213 和 DDX58 的表达在妊娠 16 天的子宫内膜中高于循环子宫内膜，并且在体内宫内输注 IFNT 后直接上调（P < 0.05）。发情后 13 天，9 个基因的子宫内膜表达增加[ARHGAP1、MGC127874、LIMS2、TBC1D1、FBXL7、C25H16orf71、LOC507810、ZSWIM4 和一个新基因（ENSBTAT00000050193）]，7 个基因的表达减少（SERBP1、SRGAP2、AL7A1、TBK1、F2RL2、MGC128929 和 WBSCR17；P < 0.05）在妊娠牛的子宫内膜中与循环牛相比。在妊娠和循环子宫内膜之间，这些 DEGs 中的 F2RL2、LIMS2、LOC507810、MGC127874、TBC1D1、WBSCR17 和 ZSWIM4 的表达差异在 16 天保持不变（P < 0.05），它们的表达均不受体内 IFNT 的直接调节。对产后奶牛血液中选定的干扰素刺激基因表达的分析显示，在人工授精后 18 天，与循环对照组相比，随后确认妊娠的奶牛中 ZXFX1、PARP12、SAMD9 和 HERC6 的表达显著增加（P < 0.05）。总之，RNAseq 在牛早期妊娠的子宫内膜中鉴定出许多新的与妊娠相关的基因，这些基因不受体内 IFNT 的调节。此外，在产后奶牛发情后 18 天，血液中直接受到体内短期 IFNT 暴露调节的一些基因的表达因妊娠状态而异。
