Multiplexed digital quantification of binge-like alcohol-mediated alterations in maternal uterine angiogenic mRNA transcriptome
Multiplexed digital quantification of binge-like alcohol-mediated alterations in maternal uterine angiogenic mRNA transcriptome
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DOI：
10.1152/physiolgenomics.00009.2012
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发表时间：
2012-06-01
影响因子：
4.6
通讯作者：
Magness, Ronald R.
中科院分区：
文献类型：
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作者：
Ramadoss, Jayanth;Magness, Ronald R.
Ramadoss J，Magness RR.酗酒样酒精介导的母体子宫血管生成mrna转录组改变的多重数字定量。Physiol Genomics 44：622-628，2012.首次发表于2012年4月24日; doi：10.1152/physiolgenomics.00009.2012.-胎儿酒精谱系障碍（FASD）的基因组研究利用全基因组微阵列/生物信息学或靶向实时PCR（RT-PCR）。我们在此首次利用了具有高通量以及专注于一个生理系统的能力的新型数字方法。本研究的目的是探讨酒精诱导的子宫血管生成相关的mRNA丰度的变化，使用数字mRNA技术。四个生物学和三个技术重复的子宫动脉内皮细胞从第三个三个月的母羊荧光激活细胞分选，验证，和治疗无或有酗酒样酒精。设计了与含有生物素的寡核苷酸共价结合的捕获探针和颜色编码的报告探针，用于85个血管生成相关基因，并使用Nanostring nCounter系统进行分析。20个基因表达下调（向下箭头）和两个上调（向上箭头），包括血管生成生长因子/受体（下箭头胎盘生长因子），粘附分子（上箭头血管生成素样-3;下箭头胶原蛋白-18A1;下箭头内皮糖蛋白），蛋白酶/基质蛋白/抑制剂（下箭头丙氨酰氨肽酶;下箭头胶原蛋白-4A3;下箭头乙酰肝素酶;下箭头纤溶酶原，上箭头纤溶酶原激活剂尿激酶;下箭头血小板因子-4;下箭头含丛蛋白结构域-1;下箭头金属蛋白酶组织抑制剂-3），转录/信号分子（下箭头心脏和神经嵴衍生物-2;下箭头DNA结合蛋白抑制剂;下箭头NOTCH-4;下箭头核糖体蛋白-L13 a1;下箭头核糖体蛋白大-P1）、细胞因子/趋化因子（下箭头白细胞介素-1B）和各种生长因子（下箭头瘦素;下箭头血小板衍生生长因子-α）;下箭头转化生长因子（TGF-α;上箭头TGF-β受体-1）。这些新的数据表明，酒精对控制血管生成的基因有显著的有害作用，支持FASD中子宫胎盘血管发育异常的机制作用。因此，三方数字基因表达系统是一个有价值的工具，可以从机制和改善的角度回答有关FASD的许多其他问题。
Ramadoss J, Magness RR. Multiplexed digital quantification of binge-like alcohol-mediated alterations in maternal uterine angiogenic mrna transcriptome. Physiol Genomics 44: 622-628, 2012. First published April 24, 2012; doi:10.1152/physiolgenomics.00009.2012.-Genomic studies on fetal alcohol spectrum disorders (FASD) have utilized either genome-wide microarrays/bioinformatics or targeted real-time PCR (RT-PCR). We utilized herein for the first time a novel digital approach with high throughput as well as the capability to focus on one physiological system. The aim of the present study was to investigate alcohol-induced alterations in uterine angiogenesis-related mRNA abundance using digital mRNA technology. Four biological and three technical replicates of uterine arterial endothelial cells from third-trimester ewes were fluorescence-activated cell sorted, validated, and treated without or with binge-like alcohol. A capture probe covalently bound to an oligonucleotide containing biotin and a color-coded reporter probe were designed for 85 angio-genesis-related genes and analyzed with the Nanostring nCounter system. Twenty genes were downregulated (down arrow) and two upregulated (up arrow), including angiogenic growth factors/receptors (down arrow placental growth factor), adhesion molecules (up arrow angiopoietin-like-3; down arrow collagen-18A1; down arrow endoglin), proteases/matrix proteins/inhibitors (down arrow alanyl aminopeptidase; down arrow collagen-4A3; down arrow heparanase; down arrow plasminogen, up arrow plasminogen activator urokinase; down arrow platelet factor-4; down arrow plexin domain containing-1; down arrow tissue inhibitor of metalloproteinases-3), transcription/signaling molecules (down arrow heart and neural crest derivatives-2; down arrow DNA-binding protein inhibitor; down arrow NOTCH-4; down arrow ribosomal protein-L13a1; down arrow ribosomal protein large-P1), cytokines/chemokines (down arrow interleukin-1B), and miscellaneous growth factors (down arrow leptin; down arrow platelet-derived growth factor-alpha); down arrow transforming growth factor (TGF-alpha; up arrow TGF-beta receptor-1). These novel data show significant detrimental alcohol effects on genes controlling angiogenesis supporting a mechanistic role for abnormal uteroplacental vascular development in FASD. The tripartite digital gene expression system is therefore a valuable tool to answer many additional questions about FASD from both mechanistic as well as ameliorative perspectives.