dre-mir-2188 targets nrp2a and mediates proper intersegmental vessel development in zebrafish embryosdre - mir - 2188目标nrp2a调节适当的节间血管发展斑马鱼胚胎.pdfVIP

Dre-miR-2188 Targets Nrp2a and Mediates Proper Intersegmental Vessel Development in Zebrafish Embryos 1 1 ´ 1 2 ´ ` 2 Ana R. Soares , Marisa Reverendo , Patrıcia M. Pereira , Olivier Nivelles , Helene Pendeville , Ana 1 1 2 1 Rita Bezerra , Gabriela R. Moura , Ingrid Struman , Manuel A. S. Santos * 1 RNA Biology Laboratory, Department of Biology CESAM, University of Aveiro, Aveiro, Portugal, 2 Unit of Molecular Biology and Genetic Engineering, GIGA-Research, ` ` University of Liege, Sart Tilman, Liege, Belgium Abstract Background: MicroRNAs (miRNAs) are a class of small RNAs that are implicated in the control of eukaryotic gene expression by binding to the 39UTR of target mRNAs. Several algorithms have been developed for miRNA target prediction however, experimental validation is still essential for the correct identification of miRNA targets. We have recently predicted that Neuropilin2a (Nrp2a), a vascular endothelial growth factor receptor which is essential for normal developmental angiogenesis in zebrafish, is a dre-miR-2188 target. Methodology: Here we show that dre-miR-2188 targets the 39-untranslated region (39UTR) of Nrp2a mRNA and is implicated in proper intersegmental vessel development in vivo. Over expression of miR-2188 in zebrafish embryos down regulates Nrp2a expression and results in intersegmental vessel disruption, while its silencing increases Nrp2a expression and intersegmental vessel sprouting. An in vivo GFP sensor assay based on a fusion between the GFP coding region and the Nrp2a 39UTR confirms that mi