Circulating microRNA as candidates for early embryonic viability in cattle.
Circulating microRNA as candidates for early embryonic viability in cattle.
复制标题
DOI：
10.1002/mrd.22856
复制
发表时间：
2017-08
影响因子：
2.5
通讯作者：
Smith MF
中科院分区：
文献类型：
--
作者：
Pohler KG;Green JA;Moley LA;Gunewardena S;Hung WT;Payton RR;Hong X;Christenson LK;Geary TW;Smith MF
携带microRNAs(MiRNAs)的血源性细胞外小泡(即外切体和微囊泡)可以作为疾病和不同生理状态(包括妊娠状态)的良好生物标志物。我们测试了一种假设，即循环细胞外小泡来源的miRNAs可以区分怀孕到第30天的奶牛和未怀孕的奶牛，或者是那些在怀孕17-30天期间出现胚胎死亡的奶牛。将36头母牛在发情期第0天(发情期)随机分为两组，每组36头，分别用可育精液和死精液进行人工授精。分别于人工授精后第0天、第17天和第24天采血。接受活精子的奶牛被追溯归类为在第30天怀孕(n=17)或在第17-30天表现出胚胎死亡(n=19)。用超速离心法从血清中分离出17天和24天的细胞外小泡，并在提取RNA之前通过纳米颗粒跟踪和蛋白质印迹分析(针对CD81)证实了它们的存在。对怀孕奶牛、胚胎死亡率奶牛和对照奶牛(每天4头)进行了microRNA测序，总共有24个独立反应。总共在血清中鉴定出214个miRNAs，其中40个是新发现的。基于差异丰度参数，我们确定了32个差异丰富的基因座，代表了27个差异丰富的成熟miRNA。在第17天和第24天，鉴定出区分妊娠状态的特定miRNAs(例如miR-25、-16b和-3596)。综上所述，我们确定了几个来自循环细胞外小泡的miRNAs，它们在胚胎死亡和怀孕奶牛之间的丰度不同。
Blood-borne extracellular vesicles (i.e. exosomes and microvesicles) carrying microRNAs (miRNAs) could make excellent biomarkers of disease and different physiologic states, including pregnancy status. We tested the hypothesis that circulating extracellular vesicle-derived miRNAs might differentiate the pregnancy status of cows that had maintained pregnancy to Day 30 from non-pregnant cows or from those that exhibited embryonic mortality between Days 17–30 of gestation. Cows were randomly assigned for artificial insemination with fertile semen (n= 36) or dead semen (n=8; control group) on Day 0 (day of estrus). Blood was collected from all animals on Day 0 and on Days 17 and 24 after artificial insemination. Cows receiving live sperm were retrospectively classified as pregnant on Day 30 (n=17) or exhibiting embryonic mortality between Days 17–30 (n=19). Extracellular vesicles from Day-17 and -24 samples were isolated from serum using ultra-centrifugation, and their presence was confirmed by nanoparticle tracking and Western blot analyses (for CD81) prior to RNA extraction. MicroRNA sequencing was performed on pregnant, embryonic-mortality, and control cows (n=4 per day), for a total of 24 independent reactions. In total, 214 miRNAs were identified in serum, 40 of which were novel. Based on differential abundance parameters, we identified 32 differentially abundant loci, representing 27 differentially abundant mature miRNA. At Days 17 and 24, specific miRNAs (e.g. miR-25, -16b and -3596) were identified that differentiated the pregnancy status. In summary, we identified several circulating extracellular vesicles derived miRNAs that differ in abundance between embryonic mortality and pregnant cows.
登录
查看更多内容
DOI：
10.1073/pnas.0808830105
发表时间：
2008-10-21
影响因子：
11.1
作者：
Kim, Daniel H.;Saetrom, Pal;Rossi, John J.
通讯作者：
Rossi, John J.
影响因子：
2.2
作者：
Dunne, LD;Diskin, MG;Sreenan, JM
通讯作者：
Sreenan, JM
影响因子：
4.6
作者：
Coutinho, Luiz L.;Matukumalli, Lakshmi K.;Smith, Timothy P. L.
通讯作者：
Smith, Timothy P. L.
DOI：
10.1073/pnas.1304718110
发表时间：
2013-07-16
影响因子：
11.1
作者：
Delorme-Axford, Elizabeth;Donker, Rogier B.;Coyne, Carolyn B.
通讯作者：
Coyne, Carolyn B.
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y