Genetic Analysis of C. elegans fat regulatory network

线虫脂肪调节网络的遗传分析

• 批准号: 6876223
• 负责人: Kaveh Ashrafi
• 金额: $ 30.3万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-20 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6876223
• 关键词: Caenorhabditis elegans; RNA interference; animal genetic material tag; appetite regulatory center; bioenergetics; biological signal transduction; carbohydrate metabolism; dietary lipid; fats; fatty acids; gene expression; gene induction /repression; green fluorescent proteins; high throughput technology; homeostasis; insulin; nutrient intake activity; polymerase chain reaction; regulatory gene; serotonin; starvation; sterols

DESCRIPTION (provided by applicant): There is mounting evidence that fat regulation involves a complex interplay between the feeding regulatory centers in the nervous system and the regulated transport, partitioning, storage, and utilization of stored fat. It is thought that this homeostatic system has evolved to ensure survival by maintaining energy reservoirs. However, in times of plentiful nutrient supplies, this homeostatic system can cause obesity in individuals with propensity for maintaining large energy stores. Obesity is associated with major heath epidemics worldwide. C. elegans provides an opportunity to identify and analyze fat regulatory genes. Importantly, these analyses are conducted in intact animals in the context of the homeostatic network. Previously, genome-wide RNAi approaches identified over 400 genes that, when inactivated, affect body fat content in C. elegans. These include components of neuroendocrine signaling, transcription, and metabolism. These findings highlight the shared ancestry of C. elegans and mammalian fat regulation and identify many novel pathways. Based on epistasis analyses and mammalian homology searches, 120 priority candidates have been identified. To understand the modes of function and regulation of these genes in intact organisms, i) feeding and locomotary rates of RNAi exposed animals will be determined, ii) quantitative RT-PCR assays will be used to monitor expression levels of a panel of fatty acid, sterol, and sugar metabolic enzymes in RNAi exposed animals, iii) genes identified to affect similar processes will be grouped into genetic pathways, iv) sites of function of fat genes and their products will be determined by GFP tag fusions, iv) these GFP-tag reporters and RT-PCR assays will be used to identify the fat gene inactivations that affect expression levels and/or localizations of the other fat regulatory genes. Similarly, impact of starvation or defects in insulin and serotonin signaling pathways on the fat regulatory genes will be assessed.

描述（由申请人提供）： 有越来越多的证据表明，脂肪调节涉及神经系统中的喂养调节中心与储存脂肪的调节，分配，储存和利用之间的复杂相互作用。人们认为，这种体内稳态系统已经发展为通过维持能源储层来确保生存。但是，在充足的营养供应时，这种稳态系统可能会导致具有维持大型能量储存倾向的人的肥胖症。肥胖与全球主要的希思流行有关。秀丽隐杆线虫提供了识别和分析脂肪调节基因的机会。重要的是，这些分析是在稳态网络的完整动物中进行的。以前，全基因组RNAi方法鉴定出超过400个基因，这些基因在灭活后会影响秀丽隐杆线虫中的体内脂肪含量。这些包括神经内分泌信号传导，转录和代谢的组成部分。这些发现突出了秀丽隐杆线虫和哺乳动物脂肪调节的共同血统，并确定了许多新颖的途径。根据上毒分析和哺乳动物同源性搜索，已经确定了120个优先级候选者。 To understand the modes of function and regulation of these genes in intact organisms, i) feeding and locomotary rates of RNAi exposed animals will be determined, ii) quantitative RT-PCR assays will be used to monitor expression levels of a panel of fatty acid, sterol, and sugar metabolic enzymes in RNAi exposed animals, iii) genes identified to affect similar processes will be grouped into genetic pathways, iv) sites of function脂肪基因及其产物将由GFP TAG融合确定，iv）这些GFP-TAG记者和RT-PCR分析将用于识别影响其他脂肪调节基因的表达水平和/或局部定位的脂肪基因失活。同样，将评估饥饿或缺陷对胰岛素和5-羟色胺信号通路对脂肪调节基因的影响。