Abstract
Amniotic fluid (AF) continuously exchanges molecules with the fetus, playing critical roles in fetal development especially via its complex components. Among these components, microRNAs are thought to be transferred between cells loaded in microvesicles. However, the functions of AF microRNAs remain unknown. To date, few studies have examined microRNAs in amniotic fluid. In this study, we employed miRCURY Locked Nucleotide Acid arrays to profile the dynamic expression of microRNAs in AF from mice on embryonic days E13, E15, and E17. At these times, 233 microRNAs were differentially expressed (p< 0.01), accounting for 23% of the total Mus musculus microRNAs. These differentially-expressed microRNAs were divided into two distinct groups based on their expression patterns. Gene ontology analysis showed that the intersectional target genes of these differentially-expressed microRNAs were mainly distributed in synapse, synaptosome, cell projection, and cytoskeleton. Pathway analysis revealed that the target genes of the two groups of microRNAs were synergistically enriched in axon guidance, focal adhesion, and MAPK signaling pathways. MicroRNA-mRNA network analysis and gene- mapping showed that these microRNAs synergistically regulated cell motility, cell proliferation and differentiation, and especially the axon guidance process. Cancer pathways associated with growth and proliferation were also enriched in AF. Taken together, the results of this study are the first to show the functions of microRNAs in AF during fetal development, providing novel insights into interpreting the roles of AF microRNAs in fetal development.
Highlights
Amniotic fluid (AF) is the fluid in the amniotic sac that appears 7–8 days after fertilization [1]
Signals from arrays at the three time-points were subjected to analysis of variance (ANOVA) analysis after normalization and this showed that 233 microRNAs were differentially expressed (p < 0.01) at E13, E15, and E17, accounting for 23% of the total Mus musculus microRNAs
The results showed that axon guidance was the most reliably enriched pathway for Uup microRNA targets (p = 1.17×10−18, False Discovery Ratio (FDR) = 1.42 ×10−15, and fold enrichment = 3.26), consistent with the Gene Ontology (GO) biological process results
Summary
AF is the fluid in the amniotic sac that appears 7–8 days after fertilization [1]. Until delivery, the growth and development of the fetus occurs in its presence. It is widely accepted that AF provides movement space and mechanical protection for the fetus [2]. A large body of evidence suggests that its components play more important roles in fetal development [3, 4]. The composition of AF reflects the dynamic status of the developing fetus, and specific.
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