The Role of Endothelin in Controlling Pharyngeal Arch Morphogenesis

描述(由申请人提供)：头面部骨骼需要协调发育过程，这些过程由许多不同的信号通路调节。这些信号通路的中断可能会导致新生儿的头面部疾病和成年人的癌症。我的项目研究了内皮素-1(Edn1)使用的分子机制，它传递了这些途径之一的信号。在Kimmel实验室研究的斑马鱼ednl突变体，在腹侧咽弓起源的骨骼结构中显示出严重的缺陷，包括腹侧咽弓软骨减少，软骨之间融合，同源异型转化，以及软骨形状的变化。Edn1所采用的信号机制尚不清楚，然而，通过正向突变筛选为Edn1途径提供线索的几个基因已经被鉴定出来，其中一个基因是plcps。Plcps突变体表现出与edn1突变体相似的颅面缺陷，支持plcps转导edn1信号促进咽弓生长和形态发生的作用。我建议1)通过测量缺乏edn1和plcp3的胚胎中的细胞增殖和细胞死亡，确定Edn1是否需要plcps来调节神经沟来源细胞的细胞增殖和细胞死亡；2)确定是否需要plcps将aPKC/PAR复合体与肌动蛋白细胞骨架的调节因子联系起来，通过破坏每个成分并进行几何形态计量分析来促进咽弓的收敛-延伸；以及3)使用正向突变筛选来确定在Edn1途径中发挥作用的新基因。这项奖学金将为我的职业生涯奠定基础，我将使用斑马鱼头面部发育模型来研究信号通路的去调节如何导致发育障碍和疾病。我的研究将提供数据，增加我们对Edn1如何启动这些发育途径的理解，这将有助于制定更好的策略来对抗癌症等疾病，当信号被干扰或去调节时，这些疾病会导致这些疾病。

DESCRIPTION (provided by applicant): The craniofacial skeleton requires the coordination of developmental processes that are regulated by a number of different signaling pathways. Disruption of these signaling pathways can lead to craniofacial disorders in newborns and cancer in adults. My project examines the molecular mechanisms employed by Endothelin-1 (Edn1) that signals one of these pathways. Zebrafish ednl mutants, studied in the Kimmel lab, exhibit severe defects in the ventral pharyngeal arch derived skeletal structures that include reduction of ventral pharyngeal arch cartilages, fusion between cartilages, homeotic transformation, and changes in cartilage shape. The signaling mechanisms employed by Edn1 remain unclear, however, several genes have been identified through forward mutagenesis screens that provide clues to the Edn1 pathway, one such gene is plcps. The plcps mutant exhibits similar craniofacial defects to ednl mutants, supporting a role for Plcps to transduce the Ednl signal to promote pharyngeal arch growth and morphogenesis. I propose to 1) determine if Ednl requires Plcps to regulate cell proliferation and cell death of neural crestderived cells, by measuring cell proliferation and cell death in embryos lacking ednl and plcp3; 2) determine if Plcps is required to link the aPKC/Par complex with regulators of the actin cytoskeleton to promote convergence-extension of the pharyngeal arches by disrupting each component and performing geometric morphometric analysis; and 3) use forward mutagenesis screens to identify new genes that function in the Ednl pathway. This fellowship will establish my career using the zebrafish craniofacial development model to investigate how the de-regulation of signaling pathways can lead to developmental disorders and disease. My research will provide data that will increase our understanding how Ednl initiates these developmental pathways, which will help create better strategies to combat diseases, such as cancer, that result when signaling is disrupted or de-regulated.