The role of primary cilia in craniofacial development

描述（由申请人提供）：体内几乎每个细胞都配备有被称为初级纤毛的膜结合的指状突起。纤毛被等同于细胞天线，检测环境中的分子信号，从而影响细胞的行为。睫状体缺陷，称为睫状体病，与广泛的人类疾病和病症有关。大量的睫状体病影响颅面复合体。面部神经嵴及其相关的骨骼成分来源于颅神经嵴细胞。在此，我将探讨如何纤毛功能在颅神经嵴细胞的个体发育。在绘制了不同发育阶段神经嵴细胞上初级纤毛的存在之后，我将通过灭活驱动蛋白-II运动亚基之一Kif 3a（初级纤毛功能所必需的蛋白质）来破坏神经嵴细胞中的初级纤毛，并确定神经嵴细胞是否能够从上皮细胞类型转变为间充质细胞、迁移、增殖和形成间充质凝聚。最后，我将通过检测分子信号并使细胞朝向该信号来确定初级纤毛是否指导神经嵴迁移。这些实验结合起来，精确地剖析了初级纤毛是如何指导形成颅骨的细胞的发育的。我的近期目标是精确定义的作用，初级纤毛神经嵴的发展，使用鸟类模型系统。我计划利用我作为博士后研究员的剩余时间，进一步探索策略，以抑制初级纤毛功能所必需的基因。1将利用我的顾问委员会的专业知识，向我介绍可应用于鸟类和斑马鱼模型系统的基因失活的替代方法。这些早期实验将在斯坦福大学丰富的学术环境中进行。我的长期目标是建立一个独立的研究计划，使用多种动物模型系统（青蛙，鱼，小鸡和老鼠）来研究颅面模式。1计划建立在我的可衡量的经验，在每个模型中，利用每个系统的好处，以解决有关物种特异性颅面发育和模式的特定问题。公共卫生相关性：初级纤毛是普遍存在的细胞器，在信号转导中发挥重要作用。初级纤毛的缺陷导致一类定义不清的缺陷，称为纤毛病。许多纤毛病变影响颅面发育。该建议的目的是探索与颅面表型纤毛病变的基础。

DESCRIPTION (provided by applicant): Almost every cell in the body is equipped with a membrane bound, finger-like projection called a primary cilium. Cilia have been equated to cellular antennae that detect molecular signals in the environment and thus influence how cells behave as a consequence. Ciliary defects, referred to as ciliopathies, are associated with a broad of human diseases and disorders. A significant number of ciliopathies affect the craniofacial complex. The facial prominences and their associated skeletal elements are derived from cranial neural crest cells. Herein, I will investigate how cilia function during the ontogeny of cranial neural crest cells. After mapping the presence of primary cilia on neural crest cells during distinct phases of development, I will disrupt primary cilia in neural crest cells by inactivating one of the kinesin-ll motor subunits, Kif3a a protein essential for primary cilia function, an determine if neural crest cells are able to transition from an epithelial cell type to a mesenchymal cell, migrate, proliferate, and form mesenchymal condensations. Finally, I will determine if primary cilia direct neural crest migration by detecting a molecular signal and orienting the cells towards that signal. Taken together these experiments precisely dissect how the primary cilia direct the development of the cells that give rise the cranial skeleton. My immediate goal is to precisely define the role of primary cilia in neural crest development using an avian model system. I plan to use my remaining time as a post-doctoral fellow to further explore strategies to inactivate genes essential for primary cilia function. 1 will utilize the expertise of my advisory committee to introduce me to the alternative approaches to gene inactivation in that can be applied to both avian and zebrafish model systems. These early experiments will be performed in the rich academic environment of Stanford University. My long term goal is to establish an independent research program that uses multiple animal model systems (frog, fish, chick, and mouse) to study craniofacial patterning. 1 plan to build on my measurable experience in each model to exploit the benefits of each system to address particular questions concerning species-specific craniofacial development and patterning. Public Health Relevance: Primary cilia are ubiquitous organelles that play vital roles in signal transduction. Defects in primary cilia are responsible for a poorly defined class of defects called ciliopathies. A significant number of ciliopathies affect craniofacial development. The goal of this proposal is to explore the basis for ciliopathies with craniofacial phenotypes.