Ribeiro E S, Monteiro A P A, Bisinotto R S, Lima F S, Greco L F, Ealy A D, Thatcher W W, Santos J E P
Department of Animal Sciences, University of Florida, Gainesville 32611; DH Barron Reproductive and Perinatal Biology Research Program, University of Florida, Gainesville 32611.
Department of Animal Sciences, University of Florida, Gainesville 32611.
J Dairy Sci. 2016 Jun;99(6):4761-4777. doi: 10.3168/jds.2015-10315. Epub 2016 Mar 24.
The objectives were to compare development and transcriptome of preimplantation conceptuses 15 d after synchronized ovulation and artificial insemination (AI) according to the genetic background of the cow and estrous cyclicity at the initiation of the synchronization program. On d 39±3 postpartum, Holstein cows that were anovular (HA; n=10), Holstein cows that were estrous cyclic (HC; n=25), and Jersey/Holstein crossbred cows that were estrous cyclic (CC; n=25) were randomly selected in a grazing herd and subjected to the Ovsynch protocol. All cows were inseminated on d 49±3 postpartum, which was considered study d 0. Blood was sampled and analyzed for concentrations of progesterone, estradiol, insulin, and insulin-like growth factor 1 (IGF-1) on study d -10, -3, -1, 7, and 15 relative to AI. On study d 15, uteri were flushed and recovered fluid had IFN-τ concentrations measured and subjected to metabolomic analysis. Morphology of the recovered conceptuses was evaluated, and mRNA was extracted and subjected to transcriptome microarray analysis. Compared with HC, CC presented greater concentrations of progesterone and estradiol in plasma, with corpora lutea and preovulatory follicles of similar size. Conceptuses from CC were larger, tended to secrete greater amounts of IFN-τ, and had greater transcript expression of peroxisome proliferator-activated receptor gamma (PPARγ), an important transcription factor that coordinates lipid metabolism and elongation at preimplantation development. In addition, pregnant CC had greater concentrations of anandamide in the uterine flush, which might be important for elongation of the conceptus and early implantation. Conceptuses from HA were also longer and secreted greater amounts of IFN-τ than conceptuses from HC, likely because of the distinct progesterone profiles before and after AI. Nonetheless, anovular cows had reduced concentrations of IGF-1 in plasma, and their conceptuses presented remarkable transcriptomic differences. Some of the altered transcripts suggest that conceptus cells from anovular cows might be under greater cellular stress and presented markers suggesting increased apoptosis and autophagy, which could lead to increased mortality after d 15 of development. Estrous cyclicity had more impact on transcriptome of bovine conceptus than genetic background, and the developmental changes observed during the preimplantation period might be linked to differences in fertility among groups.
研究目的是根据奶牛的遗传背景以及同步程序启动时的发情周期,比较在同步排卵和人工授精(AI)后15天的植入前胚胎的发育情况和转录组。产后39±3天,在一个放牧牛群中随机选择无排卵的荷斯坦奶牛(HA;n = 10)、发情周期正常的荷斯坦奶牛(HC;n = 25)以及发情周期正常的泽西/荷斯坦杂交奶牛(CC;n = 25),并对其实施Ovsynch方案。所有奶牛在产后49±3天进行人工授精,这一天被视为研究第0天。在相对于AI的研究第-10、-3、-1、7和15天采集血液样本,分析孕酮、雌二醇、胰岛素和胰岛素样生长因子1(IGF-1)的浓度。在研究第15天,冲洗子宫,测量回收液中的IFN-τ浓度并进行代谢组学分析。评估回收胚胎的形态,提取mRNA并进行转录组微阵列分析。与HC相比,CC的血浆中孕酮和雌二醇浓度更高,黄体和排卵前卵泡大小相似。CC的胚胎更大,倾向于分泌更多的IFN-τ,并且过氧化物酶体增殖物激活受体γ(PPARγ)的转录表达更高,PPARγ是一种重要的转录因子,在植入前发育过程中协调脂质代谢和胚胎伸长。此外,怀孕的CC子宫冲洗液中花生四烯酸乙醇胺浓度更高,这可能对胚胎伸长和早期着床很重要。HA的胚胎也比HC的胚胎更长,分泌的IFN-τ更多,这可能是由于AI前后孕酮水平不同。尽管如此,无排卵奶牛血浆中的IGF-1浓度降低,其胚胎呈现出显著的转录组差异。一些改变的转录本表明,无排卵奶牛的胚胎细胞可能处于更大的细胞应激状态,并呈现出表明凋亡和自噬增加的标志物,这可能导致发育15天后死亡率增加。发情周期对牛胚胎转录组的影响大于遗传背景,植入前期观察到的发育变化可能与各组之间的生育力差异有关。