The role of primary cilia in craniofacial development

几乎体内的每个细胞都有一个被膜束缚的手指状突起，称为 初级纤毛纤毛被等同于细胞的触角，可以检测细胞中的分子信号。 环境，从而影响细胞的行为。睫状体缺陷，被称为纤毛病， 与多种人类疾病有关。大量的纤毛病变影响 颅面复合体：此处。我将研究纤毛在发育过程中的作用。 颅面骨骼的前体细胞颅神经嵴在绘制了 初级纤毛神经嵴细胞在不同的发展阶段，我将利用强大的 一级纤毛破坏的鸟类遗传模型，talpid。使用Talpid，我将评估 没有初级纤毛的颅神经嵴细胞从上皮细胞转变为 间充质细胞，迁移，增殖，并形成间充质凝聚。最后要 确定初级纤毛是否通过趋化性引导神经嵴迁移。综合这些 实验将精确解剖hovv初级纤毛直接发育神经嵴细胞; IVIy的近期目标与最初声明的目标相同：准确地定义初级教育的作用， 纤毛在神经嵴发育中的作用-为此， 该提案的一部分与原始K99相同。对提案的修改来自于 用于实现特定目标的技术和试剂的形式。改革是在 对试点实验的反应，这些实验暴露了最初提议的技术局限性， 方法论例如，提出标记纤毛轴丝的一种抗体（Ar 113 b）不 在一个鸟类模型系统中工作。为了解决这个问题，我设计了一个纤毛报告器结构 在K99阶段的赠款。如果这个构造被证明是有效的，它\N\取代了以前的 列出的抗体作为睫状体标志物其他变化以不同抗体的形式出现 用于标记神经嵴细胞和用于培养神经嵴细胞的不同培养基。 我最初的长期目标是建立一个独立的研究项目， 在K99部分的资金，我接受了一个助理提供 辛辛那提儿童医院医疗中心（CCHMC）教授职位。因此 这些实验的其余部分将在CCHMC丰富的学术环境中进行， 整形外科和心理生物学系。

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. Ciliary defects, referred to as ciliopathies, are associated with a broad spectrum of human diseases. A significant number of ciliopathies affect the craniofacial complex: Herein. I vvill investigate how cilia function during the development of the precursor cells of the craniofacial skeleton, the cranial neural crest. After mapping the presence of primary cilia on neural crest cells during distinct phases of development, I will exploit the robust avian genetic model of primary cilia disruption, the talpid. Using the Talpid I will evaluate the ability of cranial neural crest cells without primary cilia to transition from an epithelial cell to a mesenchymal cell, to migrate, to proliferate, and to form mesenchymal condensations. Finally, I will determine if primary cilia direct neural crest migration by chemotaxis. Taken together these experiments will precisely dissect hovv the primary cilia direct the development of neural crest cells; IVIy immediate goal remains the same as originally stated: to preGisely define the role of primary cilia in neural Crest development using an avian model system- To this end, the sisope of the ROO portion of the proposal is identical to the original K99. The alterations to the proposal come in the form of techniques and reagents used to accomplish the specific aims. Changes were introduced in response to pilot experiments that exposed the technical limitations of the originally proposed methodology. For example, one antibody (Arl13b) proposed to mark the ciliary axoneme does not work in an avian model system. To circumvent this problem, I engineered a ciliary reporter construct during the K99 phase of the grant. If this construct proves functional, it \N\\\ replace the previously listed antibodies as a ciliary marker Additional changes come in the form of different antibodies used for marking neural crest cells and different medlas used to culture neural crest cells. My original long-term goal was to establish an independent research program to study craniofacial patterning and during the K99 portion of funding, I accepted an offer for an Assistant Professor position at the Cincinnati Children's Hospital Medical Center (CCHMC). Therefore, the remainder of these experiments will be performed in the rich academic environment of CCHMC in Departments Of Plastic Surgery and pevelopmental Biology.