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

该项目将发现胚胎发育过程中细胞内区室是如何形成的。动物细胞含有称为细胞器的隔室，通常赋予单个细胞特定的功能。其中一个区室被称为溶酶体相关细胞器（LRO），它介导重要的活动，包括身体色素的产生、营养物质的储存和衰老的信号。该项目将发现这些细胞器在动物发育过程中首次产生的时间，研究物质是如何被引导到这些隔间的，并研究它们的数量和大小是如何被调节的。作为项目的一部分，每年将有30-40名本科生在合作、调查和原创研究方面受到指导。该项目将为大批不同类型的学生提供实际的科学培训，其中大多数人可能会在科学和教育领域取得进展。目前，对于细胞分化过程中LRO的转运途径是如何出现的，内溶酶体（如晚期内溶体或溶酶体）是否在LRO蛋白递送中起作用，以及如何调节生物发生途径来控制LRO的数量和大小，我们知之甚少。为了研究这些过程，本项目分析了秀丽隐杆线虫肠道细胞中肠道颗粒，LROs的生物发生。该研究将使用遗传学方法来检查LRO生物发生基因是否在种系或胚胎中起作用，并在确定LRO蛋白在发育过程中何时首次产生后，使用定量荧光显微镜评估其相对于内体细胞器标记物的分布。此外，肠道颗粒货物是否通过内溶酶体将使用脉冲追踪和加回方法，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。通过在不同发育阶段跟踪肠道颗粒货物的靶向表达，并结合新开发的方法来潜在地破坏LRO生物发生蛋白的功能，可以确定指导LRO蛋白运输的途径何时活跃。最后，将对一组肠道颗粒增大的秀丽隐杆线虫突变体进行评估，以确定LRO稳态的潜在调节因子。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值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.