Mechanisms that maintain and remodel the sensory cilium

维持和重塑感觉纤毛的机制

• 批准号: 9889126
• 负责人: Niels Ringstad
• 金额: $ 25.43万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9889126
• 关键词: Acute; Afferent Neurons; Behavior; Binding; Biochemical; Biological Assay; Blindness; Caenorhabditis elegans; Carbon Dioxide; Cell physiology; Cell surface; Cells; Cellular Membrane; Chronic; Cilia; Cues; Cyclic GMP; Dendrites; Destinations; Development; Differentiation and Growth; Dioxygenases; Disease; Dynein ATPase; Enzymes; Esthesia; Fluorescence Microscopy; Functional disorder; Genes; Genetic; Genetic Screening; Genetic Transcription; Golgi Apparatus; Guanylate Cyclase; Homologous Gene; Hypoxia; Hypoxia Inducible Factor; In Situ; Integral Membrane Protein; Invertebrates; Left; Life; Light; Measurement; Measures; Mediating; Membrane; Metabolism; Microbe; Microtubules; Modeling; Molecular; Motor; Mutation; Nematoda; Neurons; Odors; Organelles; Pathway interactions; Photoreceptors; Physiological; Process; Proteins; Proteome; Reproduction; Resolution; Retinal Degeneration; Retinal Diseases; Retinal Dystrophy; Rhodopsin; Role; Second Messenger Systems; Sensory; Signal Transduction; Site; Stimulus; Stress; Structure; Surface; System; Testing; Transducers; Vertebrate Photoreceptors; Vesicle; atrial natriuretic factor receptor A; base; cell type; disease-causing mutation; experimental study; extracellular; hypoxia inducible factor 1; in vivo; novel; particle; photoreceptor degeneration; programs; protein transport; receptor; recruit; response; tool; trafficking; vesicle transport

PROJECT SUMMARY Sensory neurons concentrate and organize molecules used to detect environmental stimuli into cilia, which are specialized microtubule-based structures on the cell surface that function as cellular antennas. The proteins that constitute the machinery of sensory transduction are synthesized elsewhere and must be separated from other cellular proteins and transported to the cilium. The importance of mechanisms that mediate trafficking of proteins to the sensory cilium is illustrated by disease-causing mutations that disrupt this process. Mutations that compromise ciliary trafficking of the photopigment rhodopsin or the enzyme guanylyl cyclase cause retinal dystrophies marked by photoreceptor degeneration and, ultimately, blindness. Despite the importance of protein trafficking to the cilium, its underlying molecular mechanisms remain poorly understood. We propose to use chemosensory BAG neurons of the nematode C. elegans as a model for discovery of mechanisms that select and transport cargo destined for the sensory cilium. Like vertebrate photoreceptor neurons, BAG neurons use cyclic GMP as a second messenger for sensory transduction, and the enzymes and effectors that control cyclic GMP signals and turn them into electrical signals are highly similar to those found in photoreceptor neurons. Trafficking of proteins to BAG cilia can be measured in situ using high-resolution fluorescence microscopy assays, and powerful genetic tools are available to acutely or chronically manipulate specific molecular pathways in BAG neurons and determine their function in trafficking to the cilium. Importantly, C. elegans permits discovery of novel factors that mediate ciliary trafficking through genetic screens and biochemical approaches. We propose to use this powerful experimental system to (1) delineate a molecular pathway that matches cargo destined for the sensory cilium with specific motors that will carry it through the dendrite to its destination, and (2) determine how trafficking mechanisms are regulated by physiological or developmental cues that trigger remodeling of the BAG cilium. These studies will advance understanding of a cellular process that is essential for sensory neuron function and viability and will integrate cellular trafficking mechanisms with physiological and developmental programs that impact sensory cilia in vivo.

项目摘要 感觉神经元浓缩并组织用于检测环境刺激到纤毛的分子 在细胞表面上充当细胞天线的专门基于微管的结构。蛋白质 构成感官转导的机械在其他地方合成，必须与 其他细胞蛋白并运输到纤毛。调解贩运的机制的重要性 蛋白质的蛋白质是通过破坏该过程的引起疾病的突变来说明的。突变 这种损害了光化视紫红质或酶鸟叶叶尼环酶的睫毛贩运引起视网膜 具有光感受器变性的营养不良，最终是失明。尽管很重要 蛋白质运输到纤毛，其潜在的分子机制仍然很少了解。我们建议 使用线虫秀丽隐杆线虫的化学感应袋神经元作为发现机制的模型 选择和运输注定要进行感官纤毛的货物。像脊椎动物光感受器神经元一样 神经元使用环状GMP作为感官转导的第二使者，以及酶和效应子 对照环状GMP信号并将其转换为电信号，与在 光感受器神经元。可以使用高分辨率在原位测量将蛋白质贩运到纤毛袋中 荧光显微镜测定和强大的遗传工具可急性或长期操纵 袋神经元中的特定分子途径，并确定其在运输纤毛的功能。 重要的是，秀丽隐杆线虫允许发现通过遗传介导睫毛运输的新因素 屏幕和生化方法。我们建议使用此功能强大的实验系统来描绘A 与货物的货物相匹配的分子途径，这些货物与感官纤毛的货物与特定电动机的载体 通过树突到目的地，（2）确定贩运机制如何受到 触发袋纤毛重塑的生理或发育提示。这些研究将进步 了解对于感觉神经元功能和生存力至关重要的细胞过程，并将整合 具有生理和发育计划的细胞运输机制，影响感官纤毛 体内。