The Role of Endothelin in Controlling Pharyngeal Arch Morphogenesis

描述（由申请人提供）：颅面骨骼需要协调由许多不同信号通路调节的发育过程。这些信号通路的中断可导致新生儿的颅面疾病和成人的癌症。我的项目研究了内皮素-1（Edn 1）的分子机制，它是这些途径之一的信号。在Kimmel实验室研究的斑马鱼ednl突变体在腹咽弓衍生的骨骼结构中表现出严重的缺陷，包括腹咽弓软骨的减少、软骨之间的融合、同源异型转化和软骨形状的变化。Edn 1的信号传导机制尚不清楚，然而，通过正向诱变筛选已经鉴定出几个基因，这些基因为Edn 1途径提供了线索，其中一个这样的基因是plcps。plcps突变体表现出与ednl突变体相似的颅面缺陷，支持Plcps抑制Ednl信号以促进咽弓生长和形态发生的作用。我建议1）通过测量缺乏ednl和plcp 3的胚胎中的细胞增殖和细胞死亡来确定Ednl是否需要Plcps来调节神经嵴衍生细胞的细胞增殖和细胞死亡; 2）通过破坏每个组分并进行几何形态测量分析来确定Plcps是否需要连接aPKC/Par复合物与肌动蛋白细胞骨架的调节剂以促进咽弓的会聚-延伸;和3）使用正向诱变筛选来鉴定在Edn 1途径中起作用的新基因。这个奖学金将建立我的职业生涯使用斑马鱼颅面发育模型来研究信号通路的失调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Ednl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疾病，如癌症，当信号被破坏或失调时会导致疾病。

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.