Abstract
BackgroundThe human amnion plays a pivotal role in parturition. Two of its compartments, the placental amnion and the reflected amnion, have distinct transcriptome and are functionally coordinated for parturition. This study was conducted to determine the microRNA (miRNA) expression pattern and its significance in the placental amnion and the reflected amnion in association with labor at term.Methodology/Principal FindingsMicroRNA microarray, real-time quantitative RT-PCR (qRT-PCR), and miRNA in situ hybridization analyses of the placental amnion and the reflected amnion (n = 20) obtained at term were conducted. Luciferase assay, transfection, and qRT-PCR analyses of primary amnion epithelial cells (AECs) and amnion mesenchymal cells (AMCs) were performed. MicroRNA microarray analysis demonstrated differential expression of 32 miRNAs between the placental amnion and the reflected amnion after labor. Thirty-one (97%) miRNAs, which included miR-143 and miR-145, a cardiovascular-specific miRNA cluster, were down-regulated in the reflected amnion. Analyses of miR-143 and miR-145 by qRT-PCR confirmed microarray results, and further demonstrated their decreased expression in the reflected amnion with labor. Interestingly, expression of miR-143 and miR-145 was higher in AMCs than in AECs (p<0.05). Luciferase assay and transfection confirmed miR-143 binding to 3′ UTR of prostaglandin-endoperoxidase synthase 2 (PTGS2) mRNA and miR-143 regulation of PTGS2 in AMCs.ConclusionsWe report region-specific amniotic microRNAome and miR-143 regulation of PTGS2 in the context of human labor at term for the first time. The findings indicate that miRNA-mediated post-transcriptional regulation of gene expression machinery in the amnion plays an important role in the compartments (placental amnion vs reflected amnion) and in a cell type-specific manner (AECs vs AMCs) for parturition.
Highlights
Human parturition has distinct features unique from other species, and biological activation of the chorioamniotic membranes is a key component of a cascade of events leading to labor at term [1,2]
The findings indicate that miRNA-mediated post-transcriptional regulation of gene expression machinery in the amnion plays an important role in the compartments and in a cell type-specific manner (AECs vs amnion mesenchymal cells (AMCs)) for parturition
The complete microRNA microarray dataset is available in Gene Expression Omnibus (GEO; http://www.ncbi.nlm.nih.gov/geo/query/acc. cgi?token=xtebjacumgweulo&acc=GSE27441)
Summary
Human parturition has distinct features unique from other species, and biological activation of the chorioamniotic membranes is a key component of a cascade of events leading to labor at term [1,2]. The amnion is the inner layer of the chorioamniotic membranes which play critical roles in the maintenance of pregnancy and the initiation of parturition [3,4,5]. Many studies have elegantly addressed both mechanical and biochemical properties of the amnion [10,11,12,13] It is a single structure, the amnion is anatomically divided into three distinct compartments: placental amnion (amnion over the placental disc), reflected amnion (amnion of the free chorioamniotic membranes), and umbilical amnion (amnion of the umbilical cord) [14]. This study was conducted to determine the microRNA (miRNA) expression pattern and its significance in the placental amnion and the reflected amnion in association with labor at term
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