Functional analysis of draxin in cranial neural crest emigration

项目摘要 神经嵴是一种多能干细胞样细胞群，是脊椎动物所特有的， 正常的颅面形态发生这些细胞有助于产生各种各样的衍生物，包括 颅面骨骼和软骨，周围神经系统的感觉和自主神经节，以及 皮肤的色素沉着。神经嵴祖细胞出现在神经板边缘，在神经和非神经板之间。 神经外胚层，并且在神经形成后，位于未来中枢神经系统的背侧， 神经管然后，神经嵴细胞经历上皮向间充质转化（EMT），以离开神经嵴。 神经管，有时会广泛迁移到远处。一旦他们到达目的地，神经 嵴细胞停止它们的迁移程序并分化成广泛的衍生物。神经嵴是 对于面部的发育是必不可少的-影响神经嵴生成，迁移， 增殖或分化导致影响面部的许多疾病和畸形，例如Treacher 柯林斯综合征，迪乔治综合征，腭裂等等。因此，需要有基本的 关于神经嵴发育的精确机制的科学知识。 为此，本提案的具体目标是利用功能丧失实验： 1)通过以下方式表征基因draxin在颅神经嵴命运特化和EMT/迁移中的作用： RNA测序和延时共聚焦显微镜;以及 2)确定Draxin与之相互作用以调节颅神经嵴发育的信号通路， 通过荧光报告表达和原位蛋白质-蛋白质相互作用测定。 本提案的目标是确定Draxin调节颅神经嵴的机制 发展过程中的EMT和迁移。因此，本提案的结果将大大提高我们的 了解颅神经嵴EMT和迁移的调节，为研究颅神经嵴EMT和迁移提供新的科学途径。 颅面缺损治疗的转化研究应用。

PROJECT SUMMARY The neural crest is a multipotent stem cell-like population, unique to vertebrates, that is essential for normal craniofacial morphogenesis. These cells contribute to a wide variety of derivatives, including the craniofacial skeleton and cartilage, sensory and autonomic ganglia of the peripheral nervous system, and pigmentation of the skin. Neural crest progenitors arise at the neural plate border, between neural and non- neural ectoderm, and, after neurulation, reside within the dorsal aspect of the future central nervous system, the neural tube. Neural crest cells then undergo an epithelial-to-mesenchymal transition (EMT) to exit the neural tube, and migrate extensively at times to distant locations. Once they reach their destination, neural crest cells cease their migratory program and differentiate into a wide range of derivatives. The neural crest is indispensable for the development of the face— genetic defects affecting neural crest generation, migration, proliferation, or differentiation, result in numerous diseases and malformations affecting the face, e.g. Treacher Collins syndrome, DiGeorge syndrome, and cleft palate to name a few. Thus, there is a need for basic scientific knowledge regarding the precise mechanisms underlying neural crest development. To this end, the Specific Aims of this Proposal seek to use loss-of-function experiments to: 1) Characterize the role of a gene, draxin, in cranial neural crest fate specification and EMT/migration by RNA sequencing and time-lapse confocal microscopy; and 2) Identify the signaling pathway(s) with which draxin interacts to regulate cranial neural crest development, by fluorescent reporter expression and in situ protein-protein interaction assays. The goal of this Proposal is to identify the mechanism by which draxin regulates cranial neural crest EMT and migration during development. Thus, the results of this Proposal will significantly enhance our understanding of the regulation of cranial neural crest EMT and migration, providing new scientific avenues for translational research applications in the treatment of craniofacial defects.