Molecular Determinants of Vertebrate Vasculature and Development

背景：脊椎动物的血管系统对血液循环、气体miRNA功能至关重要；我们用Ep4激动剂刺激HUVEC细胞的miRNA，用Ep4拮抗剂抑制HUVEC-miR细胞的miRNA功能。目的：(A)研究PGE2在斑马鱼胚胎发育和血管生成中的作用；将PGE2以剂量依赖的方式注射到鱼类胚胎中，并记录发育变化。我们将用胚胎提取物进行微阵列来识别miRNA的变化。(二)为了测试miRNA的功能，我们将敲除斑马鱼胚胎中的miRNA，并测量生长变化。这项初步研究将发现在血管形成过程中受PGE2调控的新的miRNAs。目标4：发现PGE2处理后斑马鱼基因表达的变化；我们将在载体上进行微阵列和PGE2处理的鱼胚胎提取的mRNAs，以确定可能参与脊椎动物发育的新基因。意义：因此，该计划将通过各种方法，在脊椎动物发育的目录中增加新的决定因素。鉴定PGE2和miRNA在血管生成中的生理作用；Ep4信号调节miRNA功能，因此，该计划将修改目前使用的针对Ep4的药物的进程。

Background: Vertebrate vascular system is vital for blood circulation, gas miRNA functions; we shall stimulate miRNA in HUVEC cells with EP4 agonist and inhibit miRNA functions in HUVEC-miR cells with EP4 antagonist. Functional dependence on EP4 mediated signaling (PI3K/AKT, ERK phosphorylation) will establish the link between miRNAs and EP4.Objective 3: (a) To study the role of the PGE2 in embryonic development and vasculogenesis in Zebrafish model; we shall inject PGE2 in fish embryo in a dose-dependent way and record developmental changes. We shall conduct microarrays with embryo extracts to identify miRNA changes. (b) To test miRNA functions, we shall knock down miRNAs in Zebrafish embryo and measure growth change. This pilot study will identify novel miRNAs regulated by PGE2 during vasculogenesis.Objective 4: To find gene expression changes due to PGE2 treatment in Zebrafish; we shall conduct microarrays in Vehicle and PGE2 treated fish embryo extracted mRNA to identify novel genes might involve in vertebrate development.Significance: With various approaches, this program will, therefore, add new determinants in the catalog of vertebrate development. Identification of the physiological roles of PGE2 and miRNA in vasculogenesis; EP4 signaling regulating miRNA functions, this program will, therefore, modify the course of currently used drugs targeting EP4.