Understanding Ciliary Functions in Mammalian Development

项目摘要/摘要 许多细胞的表面有被称为纤毛的突起。小鼠的遗传学在 揭示了对发育中的纤毛的新要求，如左/右轴规范，以及骨骼和 神经管构图。在哺乳动物的发育过程中，纤毛产生和感觉流动，并解释 细胞间的信号，例如刺猬的信号。人类的睫毛功能缺陷会导致多种疾病 被称为纤毛病。尽管纤毛很重要，但纤毛生物学的基本方面，包括 纤毛是如何构造的，它们是如何发出信号的，以及它们是如何协调不同的发育事件的，这些问题仍然存在 人们对此知之甚少。 为了阐明这些长期存在的问题的答案，我们建议利用一个独特的资源， 这些基因敲除的小鼠突变株是由国际小鼠表型联盟(IMPC)创造的。 我们开发了一种创新的算法，并证明了它可以识别纤毛 分析将是有价值的。我们将使用这个算法来选择突变体，并用这些突变体回答三个问题 关于纤毛的补充问题： 1)纤毛的复杂结构和亚区是如何构建的，并对纤毛功能有何贡献？ 2)纤毛运动是如何建立和调节的？ 3)纤毛如何转导细胞内信号，如刺猬信号，以及这种信号的丢失是如何 细胞间通讯在纤毛疾病的发病机制中起作用？ 为了解决这些问题，我们将结合先进的成像方法，包括超分辨率 显微镜，微型计算机断层扫描，使用新的遗传工具，如用绿色荧光蛋白标记纤毛的转基因。 在我们总共25年的老鼠纤毛开发工作中积累的专业知识将使我们能够 使用新的小鼠突变体来揭示纤毛发生和纤毛信号的新原理。这些 这些发现将有助于确定孤儿疾病的致病基因，并阐明其发育起源。 人类纤毛病。

Project Summary/Abstract Many cells possess projections from their surfaces called cilia. Mouse genetics has been instrumental in uncovering novel requirements for cilia in development, such as left/right axis specification, and skeletal and neural tube patterning. During mammalian development, cilia generate and sense flow, and interpret intercellular cues, such as Hedgehog signals. Defects in ciliary function in humans cause diverse diseases known as ciliopathies. Despite the importance of the cilium, fundamental aspects of ciliary biology, including how cilia are constructed, how they signal, and how they coordinate diverse developmental events, remain poorly understood. To illuminate answers to these longstanding questions, we propose to make use of a unique resource, the knockout mouse mutant strains being created by the International Mouse Phenotyping Consortium (IMPC). We have developed an innovative algorithm and demonstrated that it can identify mutant lines for which ciliary analysis will be valuable. We will use this algorithm to select mutants, and use these mutants to answer three complementary questions about cilia: 1) How are the sophisticated structures and subdomains of cilia built and contribute to ciliary function? 2) How is ciliary motility established and regulated? 3) How do cilia transduce intracellular signals, such as Hedgehog signals, and how does loss of this intercellular communication contribute to the pathogenesis of ciliopathies? To address these questions, we will combine advanced imaging approaches, including super-resolution microscopy, micro-computed tomography, with novel genetic tools such as transgenes that label cilia with GFP. The expertise accrued during our combined 25 years working on cilia in mouse development will allow us to use the novel mouse mutants to uncover novel principles underlying ciliogenesis and ciliary signaling. These discoveries will help identify the causative genes for orphan diseases, and illuminate the developmental origins of human ciliopathies.