pH Regulation of Fat Storage in the Nematode Intestine

线虫肠道脂肪储存的 pH 调节

• 批准号: 6952415
• 负责人: Keith Nehrke
• 金额: $ 31.2万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-27 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6952415
• 关键词: Caenorhabditis elegans; RNA interference; Xenopus; acid base balance; acidosis; biological signal transduction; cell line; cholesterol; dietary restriction; fluorescence microscopy; functional /structural genomics; gastrointestinal absorption /transport; gene deletion mutation; genetic mapping; genetic screening; glucose metabolism; lipid metabolism; longevity; membrane transport proteins; microarray technology; mitochondria; oxygen consumption; protein isoforms; protein localization; recombinant proteins; sodium hydrogen exchanger; suppressor mutations

DESCRIPTION (provided by applicant): Deficiencies in Na+/H+ or CI-/HCO3- exchange compromise fluid and electrolyte transport in the mammalian intestine, but the ultimate role of acid-base transport in nutrient uptake processes and lipid metabolism is relatively poorly understood. In addition, little is known regarding how intestinal pHi influences cell signaling events that response to nutrient availability. We hypothesis that pHi acts as a synergistic messenger, and provides a metabolic context through which the actions of diverse cellular and trans-cellular signaling pathways are interpreted. In this application, we offer preliminary evidence linking acid-base transport to fat storage and related signaling events in the nematode C. elegans, and propose the following: Aim 1 involves characterizing the SLC4 and SLC26 CI-/HCO3- homologs in C. elegans by first assessing their distribution, then examining function both in situ, using recombinant protein expression and in vivo, using a genetically encoded pH indicator and RNAi. Aim 2 is designed to test how changes in intestinal pHi affect classic cell signaling processes that regulate fat storage, and conversely what cell signaling processes are involved in conveying the phenotypic effects of cellular acidosis. Aim 3 is designed to utilize genetic, reverse-genetic, and functional genomic assays to identify regulatory proteins that attenuate intracellular pH, as well as the global transcriptional responses to intracellular acidification. The strength of this proposal is that it will use a powerful model system and novel reagents for monitoring pH in living organisms to study fat biology from a unique perspective: that of acid-base homeostasis.

描述（由申请人提供）： Na+/H+或CI-/HCO3-交换中的缺陷会损害哺乳动物肠中的液体和电解质转运，但是酸碱转运在养分摄取过程中的最终作用和脂质代谢相对较众所周知。此外，关于肠道PHI如何影响对养分可用性的响应的细胞信号事件，知之甚少。我们假设PHI充当协同的使者，并提供了一种代谢环境，通过该环境可以解释各种细胞和跨细胞信号传导途径的作用。 In this application, we offer preliminary evidence linking acid-base transport to fat storage and related signaling events in the nematode C. elegans, and propose the following: Aim 1 involves characterizing the SLC4 and SLC26 CI-/HCO3- homologs in C. elegans by first assessing their distribution, then examining function both in situ, using recombinant protein expression and in vivo, using a genetically encoded pH indicator and RNAi. AIM 2旨在测试肠道PHI的变化如何影响调节脂肪储存的经典细胞信号传导过程，相反，在传达细胞型酸中毒的表型效应中涉及哪些细胞信号过程。 AIM 3旨在利用遗传，反遗传学和功能基因组测定方法来鉴定减弱细胞内pH的调节蛋白，以及对细胞内酸化的全局转录反应。该提案的优势在于，它将使用强大的模型系统和新型试剂来监测活生物体中的pH值，从独特的角度研究脂肪生物学：酸碱稳态的pH。