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雌二醇对牛滋养层和子宫基因转录本在妊娠母体识别周围的影响†。

Influence of estradiol on bovine trophectoderm and uterine gene transcripts around maternal recognition of pregnancy†.

机构信息

Department of Animal Science, South Dakota State University, Brookings, SD, USA.

USDA, Agricultural Research Service, Roman L. Hruska US Meat Animal Research Center, Clay Center, NE, USA.

出版信息

Biol Reprod. 2021 Aug 3;105(2):381-392. doi: 10.1093/biolre/ioab091.

Abstract

Embryo survival and pregnancy success is increased among animals that exhibit estrus prior to fixed time-artificial insemination, but there are no differences in conceptus survival to d16. The objective of this study was to determine effects of preovulatory estradiol on uterine transcriptomes, select trophectoderm (TE) transcripts, and uterine luminal fluid proteins. Beef cows/heifers were synchronized, artificially inseminated (d0), and grouped into either high (highE2) or low (lowE2) preovulatory estradiol. Uteri were flushed (d16); conceptuses and endometrial biopsies (n = 29) were collected. RNA sequencing was performed on endometrium. Real-time polymerase chain reaction (RT-PCR) was performed on TE (n = 21) RNA to measure relative abundance of IFNT, PTGS2, TM4SF1, C3, FGFR2, and GAPDH. Uterine fluid was analyzed using 2D Liquid Chromatography with tandem mass spectrometry-based Isobaric tags for relative and absolute quantitation (iTRAQ) method. RT-PCR data were analyzed using the MIXED procedure in SAS. There were no differences in messenger RNA (mRNA) abundances in TE, but there were 432 differentially expressed genes (253 downregulated, 179 upregulated) in highE2/conceptus versus lowE2/conceptus groups. There were also 48 differentially expressed proteins (19 upregulated, 29 downregulated); 6 of these were differentially expressed (FDR < 0.10) at the mRNA level. Similar pathways for mRNA and proteins included: calcium signaling, protein kinase A signaling, and corticotropin-releasing hormone signaling. These differences in uterine function may be preparing the conceptus for improved likelihood of survival after d16 among highE2 animals.

摘要

在固定时间人工授精前表现发情的动物中,胚胎存活率和妊娠成功率增加,但到 d16 的胚胎存活率没有差异。本研究的目的是确定促排卵雌二醇对子宫转录组的影响,选择滋养外胚层(TE)转录本,并检测子宫腔液蛋白。牛/小母牛被同步发情,人工授精(d0),并分为高(高 E2)或低(低 E2)促排卵雌二醇组。在 d16 冲洗子宫;收集胚胎和子宫内膜活检(n=29)。对子宫内膜进行 RNA 测序。对 TE(n=21)RNA 进行实时聚合酶链反应(RT-PCR),以测量 IFNT、PTGS2、TM4SF1、C3、FGFR2 和 GAPDH 的相对丰度。使用二维液相色谱与串联质谱基于等重标记相对和绝对定量(iTRAQ)方法分析子宫液。使用 SAS 中的 MIXED 过程分析 RT-PCR 数据。TE 中的信使 RNA(mRNA)丰度没有差异,但在高 E2/胚胎与低 E2/胚胎组中,有 432 个差异表达基因(253 个下调,179 个上调)。还有 48 个差异表达蛋白(19 个上调,29 个下调);其中 6 个在 mRNA 水平上差异表达(FDR<0.10)。mRNA 和蛋白质的相似途径包括:钙信号、蛋白激酶 A 信号和促肾上腺皮质激素释放激素信号。这些子宫功能的差异可能是为了在高 E2 动物中提高 d16 后胚胎的存活可能性做准备。

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