Ribeiro Eduardo S, Greco Leandro F, Bisinotto Rafael S, Lima Fábio S, Thatcher William W, Santos José E
DH Barron Reproductive and Perinatal Biology Research Program, Department of Animal Sciences, University of Florida, Gainesville, Florida.
DH Barron Reproductive and Perinatal Biology Research Program, Department of Animal Sciences, University of Florida, Gainesville, Florida
Biol Reprod. 2016 Apr;94(4):97. doi: 10.1095/biolreprod.115.134908. Epub 2016 Mar 2.
The objectives of this study were to characterize changes in transcriptome of preimplantation conceptuses at the onset of elongation and associated changes in uterine histotroph composition and endometrial physiology. Lactating dairy cows (n = 160) had their ovulation synchronized by artificial insemination (study Day 0). On Day 15, uteri were flushed and endometrium tissue collected. Recovered conceptuses were classified based on morphology/length as ovoid (1-4 mm), tubular (5-19 mm), and filamentous (20-60 mm). Total RNA (n = 48) was subjected to transcriptome analysis. The uterine fluid (n = 30) was evaluated by mass spectrophotometry. Transcriptome analyses revealed drastic changes in the transition from ovoid to tubular and from tubular to filamentous. Differentially expressed genes were associated with cellular movement, cell-to-cell signaling, cellular assembly and organization, lipid metabolism, small molecule biochemistry, and molecular transport. Specific changes included reorganization of cytoskeleton and cell migration, arginine metabolism, growth factors signaling, and lipid metabolism. Functional analysis revealed fatty acids and peroxisome proliferator activated receptor gamma as upstream regulators of transcriptome changes. Expression of PPARG increased 17-fold during the onset of elongation and was highly correlated with genes involved in lipid metabolism. The histotroph was rich in amino acids, lipids, saccharides, and other intermediate metabolites, and important changes in composition occurred in the presence of a conceptus. Pregnancy had a major impact on the concentrations of important lipids in the uterine fluid and expression of genes in the endometrium. Collectively, conceptus elongation involves remarkable changes in transcriptome, composition of the histotroph, and endometrial physiology, which help elucidate important events in uterine and conceptus biology at the onset of elongation.
本研究的目的是描述着床前胚胎在伸长开始时转录组的变化,以及子宫组织营养成分和子宫内膜生理学的相关变化。泌乳奶牛(n = 160)通过人工授精使其排卵同步(研究第0天)。在第15天,冲洗子宫并收集子宫内膜组织。根据形态/长度将回收的胚胎分为卵形(1 - 4毫米)、管状(5 - 19毫米)和丝状(20 - 60毫米)。对总RNA(n = 48)进行转录组分析。通过质谱分析法评估子宫液(n = 30)。转录组分析揭示了从卵形到管状以及从管状到丝状转变过程中的剧烈变化。差异表达基因与细胞运动、细胞间信号传导、细胞组装和组织、脂质代谢、小分子生物化学以及分子运输有关。具体变化包括细胞骨架的重组和细胞迁移、精氨酸代谢、生长因子信号传导以及脂质代谢。功能分析表明脂肪酸和过氧化物酶体增殖物激活受体γ是转录组变化的上游调节因子。在伸长开始时,PPARG的表达增加了17倍,并且与参与脂质代谢的基因高度相关。组织营养富含氨基酸、脂质、糖类和其他中间代谢产物,并且在有胚胎存在的情况下其组成发生了重要变化。妊娠对子宫液中重要脂质的浓度以及子宫内膜中基因的表达有重大影响。总体而言,胚胎伸长涉及转录组、组织营养成分和子宫内膜生理学的显著变化,这有助于阐明伸长开始时子宫和胚胎生物学中的重要事件。