Understanding Ciliary Functions in Mammalian Development

项目总结/摘要 许多细胞表面都有突起，称为纤毛。小鼠遗传学在 揭示了纤毛在发育中的新要求，例如左/右轴规格，以及骨骼和 神经管模式在哺乳动物的发育过程中，纤毛产生并感知水流， 细胞间信号，如刺猬信号。人类睫状体功能的缺陷会导致多种疾病 也就是纤毛病尽管纤毛的重要性，纤毛生物学的基本方面，包括 纤毛是如何构造的，它们如何发出信号，以及它们如何协调不同的发育事件， 不太了解。 为了阐明这些长期存在的问题的答案，我们建议利用一种独特的资源， 敲除小鼠突变株由国际小鼠表型鉴定协会（IMPC）创建。 我们已经开发了一种创新的算法，并证明它可以识别突变株系， 分析是有价值的。我们将用这个算法来选择突变体，并用这些突变体来回答三个 关于纤毛的补充问题： 1)纤毛的复杂结构和亚结构域是如何构建的，并对纤毛功能起作用的？ 2)纤毛运动是如何建立和调节的？ 3)纤毛是如何传递细胞内信号的，如刺猬信号，以及这种信号的丢失是如何影响细胞内信号的？ 细胞间通讯有助于纤毛病变的发病机制？ 为了解决这些问题，我们将结合联合收割机先进的成像方法，包括超分辨率 显微镜，微计算机断层扫描，与新的遗传工具，如转基因标记纤毛与绿色荧光蛋白。 在我们25年来共同研究小鼠纤毛发育的过程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传导的新原理。这些 这些发现将有助于确定孤儿病的致病基因，并阐明孤儿病的发育起源。 人类纤毛病的症状

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.