Functional analysis of draxin in cranial neural crest emigration

项目总结 神经脊是一种多能干细胞样群，是脊椎动物所特有的，对于 正常的头面部形态发育。这些细胞参与了各种各样的衍生品，包括 颅面骨骼和软骨，周围神经系统的感觉神经节和自主神经节，以及 皮肤的色素沉着。神经脊祖细胞出现在神经板的交界处，介于神经和非神经之间 神经外胚层，在神经形成后，驻留在未来中枢神经系统的背侧， 神经管。然后神经脊细胞经历上皮到间充质的转变(EMT)以退出 神经管，有时会广泛迁移到遥远的地方。一旦他们到达目的地，神经质 CREST细胞停止其迁移程序，并分化为广泛的衍生品。神经峰是 面部发育所必需的遗传缺陷影响神经脊的产生，迁移， 增殖或分化会导致许多影响面部的疾病和畸形，例如Treacher 柯林斯综合征、迪乔治综合征和腭裂是其中的几个例子。因此，有必要建立基本的 关于神经脊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)通过以下方法来表征基因draxin在脑神经脊命运指定和EMT/迁移中的作用 RNA测序和时间推移共聚焦显微镜；以及 2)确定德拉欣与之相互作用调节脑神经脊线发育的信号通路(S)， 通过荧光报告表达和原位蛋白质-蛋白质相互作用分析。 这项建议的目的是确定德拉新调节脑神经脊线的机制。 开发过程中的EMT和迁移。因此，这项提议的结果将大大增强我们的 了解颅神经脊EMT和迁移的规律，为临床提供新的科学途径 翻译学研究在颅面骨缺损治疗中的应用。

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.