The role of primary cilia in craniofacial development

几乎体内的每个细胞都配备了一个被膜捆绑的手指状突起，称为 初级纤毛。纤毛被等同于细胞触角，它探测细胞中的分子信号 环境，从而影响细胞的行为。纤毛缺陷，称为纤毛病变，是 与广泛的人类疾病有关。相当数量的纤毛病会影响 颅面复合体：在此。我将研究纤毛如何在细菌的发育过程中发挥作用 颅面骨架的前体细胞，也就是脑神经脊。在映射存在之后 初级纤毛在神经脊细胞发育的不同阶段，我会开发出稳健的 原生纤毛破坏的禽类遗传模型。使用塔尔比德，我将评估能力 无初生纤毛的脑神经脊细胞从上皮细胞转变为 间充质细胞，迁移，增殖，形成间充质凝集。最后，我会 确定初级纤毛是否通过趋化作用引导神经脊迁移。把这些放在一起 实验将精确解剖HOVV的原生纤毛，指导神经脊细胞的发育； IVIIi近期目标保持不变：预先定义主要的角色 使用鸟类模型系统的神经脊发育中的纤毛-为此，Roo的女性化 提案的一部分与原来的K99完全相同。对提案的修改来自于 用于实现特定目的的技术和试剂的形式。在以下方面引入了更改 对暴露了最初提议的技术限制的试点实验的回应 方法论。例如，一种被提议标记睫状轴丝的抗体(Arl13b)不能 在鸟类模型系统中工作。为了绕过这个问题，我设计了一个纤毛记者构造 在资助的K99阶段。如果此构造被证明是有效的，则它\N\替换以前的 作为纤毛标志物列出的抗体以不同抗体的形式出现的其他变化 用于标记神经脊细胞和用于培养神经脊细胞的不同培养液。 我最初的长期目标是建立一个独立的研究项目来研究 在K99部分的资助中，我接受了一名助理的工作 辛辛那提儿童医院医学中心教授职位。因此， 其余的实验将在CCHMC#年丰富的学术环境中进行。 整形外科和发育生物学系。

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.