RUI: Investigating the pathways and regulation of lysosome-related organelle biogenesis.

该项目将发现细胞内隔间是如何在胚胎发育过程中形成的。动物细胞包含被称为细胞器的隔间，这些细胞器通常赋予单个细胞特殊的功能。其中一个隔间被称为溶酶体相关细胞器(LRO)，它调节重要的活动，包括身体色素的产生，营养物质的储存，以及衰老信号。这个项目将发现这些细胞器是在动物发育过程中第一次产生的，调查材料是如何被引导到这些隔室的，并研究它们的数量和大小是如何调节的。每年，作为项目的一部分，30-40名本科生将在合作、调查和原创研究方面得到指导。该项目将为一大群不同的学生提供实际操作的科学培训，他们中的大多数可能会在科学和教育领域取得进展。目前人们对LRO的运输途径在细胞分化过程中如何出现，内溶体(如晚期内切体或溶酶体)是否在LRO蛋白质传递中发挥作用，以及生物发生途径如何调控LRO的数量和大小知之甚少。为了研究这些过程，该项目分析了肠道颗粒的生物发生，即在线虫肠道细胞中发现的LRO。这项研究将使用遗传学方法来检查LRO生物发生基因是否在生殖系或胚胎中发挥功能，并在确定LRO蛋白在发育过程中第一次产生的时间后，使用定量荧光显微镜评估它们相对于内体细胞器标记的分布。此外，肠道颗粒货物是否通过内溶酶体路由，将使用脉冲追逐和加回方法，以及内溶酶体和肠道颗粒之间相互作用的活细胞荧光成像。当引导LRO蛋白运输的途径活跃时，将通过跟踪其在不同发育阶段的靶向表达后的肠道颗粒货物，结合新开发的有效干扰LRO生物发生蛋白功能的方法来确定。最后，将对一组具有增大肠道颗粒的线虫突变体进行评估，以确定LRO自我调节的潜在调节者。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The project will discover how intracellular compartments are formed during embryonic development. Animal cells contain compartments called organelles that often gives individual cells their specialized function. One of these compartments, called a lysosome-related organelle (LRO), mediates important activities, including the creation of body pigments, the storage of nutrients, and signals for aging. This project will discover when these organelles are first created during animal development, investigate how material is directed to these compartments, and study how their number and size are regulated. Each year, 30-40 undergraduates will be mentored in collaborative, investigative, and original research as part of the project. The project will provide hands on scientific training for a large and diverse group of students, most of who will likely progress on to careers in science and education.Little is currently known about how the trafficking pathways to LROs emerge during cellar differentiation, whether endolysosomes, such as late endosomes or lysosomes, function in LRO protein delivery, and how biogenesis pathways are regulated to control LRO number and size. To investigate these processes, this project analyzes the biogenesis of gut granules, LROs found in C. elegans intestinal cells. The research will use genetic approaches to examine whether LRO biogenesis genes function in the germline or embryo, and after determining when LRO proteins are first made during development, evaluate their distribution relative to endosomal organelle markers using quantitative fluorescence microscopy. In addition, whether gut granule cargo is routed through endolysosomes will be using pulse chase and add-back approaches, as well as live-cell fluorescence imaging of interactions between endolysosomes and gut granules. When pathways directing LRO protein trafficking are active will be determined by tracking gut granule cargo following its targeted expression at different stages of development combined with newly developed methods to potently disrupt the function of LRO biogenesis proteins. Finally, a collection of C. elegans mutants with enlarged gut granules will be assessed to identify potential regulators of LRO homeostasis.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.