Cellular and Genetic Analysis of Lysosome and Lysosome-Related Organelle Biogenesis in C. Elegans

线虫溶酶体和溶酶体相关细胞器生物发生的细胞和遗传分析

• 批准号: 0716280
• 负责人: Greg Hermann
• 金额: $ 36.5万
• 依托单位: Lewis and Clark College
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2011-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0716280&HistoricalAwards=false
• 关键词: Cellular; Genetic; Analysis; Lysosome; Related

Lysosomes are membrane bound compartments found in nearly all animal cells that have well characterized roles in the degradation of intra and extra-cellular derived material. While much has been learned regarding the diverse and important processes that lysosomes carry out in cells, the mechanisms underlying the assembly of lysosomes in complex multicellular animals, especially as they develop during embryogenesis, are relatively unstudied. To aid in understanding of these processes, Dr. Hermann is identifying and characterizing genes necessary for the assembly of lysosomes in the embryonic intestine of the model multicellular organism C. elegans. The Hermann lab has identified gut granules as a cell type specific, lysosome-related organelle that functions in the storage of fat. To gain detailed insight into the mechanisms controlling the formation of gut granules the Hermann lab has isolated and is characterizing a collection of glo mutants that are defective in gut granule biogenesis. Most genes identified to date that function in the biogenesis of lysosome-related organelles are not required for embryonic viability. The Hermann lab has identified 22 embryonic lethal glo mutants that extracellularly mislocalize gut granule contents. The genes necessary for the formation of gut granules whose activity is disrupted in these strains will be molecularly identified. Studies of these genes are likely to provide novel insights into the pathways necessary for lysosome biogenesis in multicellular organisms. Recent studies in C. elegans suggest that yolk platelets and LMP-1::GFP containing compartments in the intestinal primordium also represent lysosome-related organelles. Whether LMP-1::GFP containing organelles, yolk platelets, and gut granules have characteristics of conventional lysosomes, other endosomal compartments, or each other will be investigated using microscopic and genetic approaches. If these three compartments represent distinct lysosome-related organelles, the embryonic intestine would provide a powerful genetic system to investigate how structurally and functionally distinct lysosomal compartments are formed in the same cell.The project will involve 25 to 30 undergraduates a year in independent, hands-on, original research. Undergraduate students working in the PI's lab and upper division Cell Biology course (enrollment 24) will carry out the studies. The work will provide molecular insights into the assembly of lysosomes likely to be conserved in the assembly/maintenance of lysosomes in all multicellular animals.

溶酶体是几乎所有动物细胞中发现的膜结合区室，其在细胞内和细胞外衍生物质的降解中具有明确的作用。 虽然人们对溶酶体在细胞中进行的各种重要过程已经了解很多，但复杂多细胞动物中溶酶体组装的机制，尤其是在胚胎发生过程中的发育，相对而言尚未被研究。 为了帮助理解这些过程，赫尔曼博士正在鉴定和表征模型多细胞生物秀丽隐杆线虫胚胎肠道中溶酶体组装所必需的基因。 赫尔曼实验室已确定肠道颗粒是一种细胞类型特异性、与溶酶体相关的细胞器，具有储存脂肪的功能。 为了详细了解控制肠道颗粒形成的机制，赫尔曼实验室分离并鉴定了一系列在肠道颗粒生物发生方面存在缺陷的 glo 突变体。迄今为止发现的大多数在溶酶体相关细胞器的生物发生中起作用的基因并不是胚胎活力所必需的。 Hermann 实验室已鉴定出 22 个胚胎致死性 glo 突变体，它们会在细胞外使肠道颗粒内容物发生错误定位。 肠道颗粒形成所需的基因在这些菌株中被破坏，这些基因的活性被破坏。 对这些基因的研究可能为多细胞生物中溶酶体生物发生所需的途径提供新的见解。 最近对秀丽隐杆线虫的研究表明，肠原基中含有卵黄血小板和 LMP-1::GFP 的区室也代表了溶酶体相关的细胞器。 将使用显微镜和遗传学方法研究含有 LMP-1::GFP 的细胞器、卵黄血小板和肠道颗粒是否具有传统溶酶体、其他内体区室或彼此的特征。 如果这三个区室代表不同的溶酶体相关细胞器，那么胚胎肠道将提供一个强大的遗传系统来研究结构和功能不同的溶酶体区室是如何在同一细胞中形成的。该项目每年将有 25 至 30 名本科生参与独立、实践、原创的研究。 在 PI 实验室和高年级细胞生物学课程（24 名招生）工作的本科生将进行研究。 这项工作将为所有多细胞动物中溶酶体的组装/维护中可能保守的溶酶体组装提供分子见解。