Minority Predoctoral Fellowship Program

少数族裔博士前奖学金计划

基本信息

批准号：
7636846
负责人：
DAVID W JOHNSON
金额：
$ 4.12万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-01 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7636846
关键词：
Address Affect Aging Animal Model Biological Assay Biological Models Biosensor Caenorhabditis elegans Caloric Restriction Cell physiology Cells Code Communication Complex Coupling Data Defect Deletion Mutation Diabetes Mellitus Disease Dyes Electron Transport Embryo Encephalopathies Fatty acid glycerol esters Fellowship Program Functional disorder Gene Targeting Genes Genetic Genetic Epistasis Genetic Models Health Heart Diseases Human Imaging Techniques In Vitro Intestines Lead Leigh Disease Lethal Genes Life Link Longevity Mammals Measures Metabolic Minority Mitochondria Morphology Muscle Mutation Nematoda Neurons Nutrient Obesity Organelles Orthologous Gene Oxidation-Reduction Oxidative Phosphorylation Pattern Pharmacology Phenotype Physiology Population Process Production Proteins Quantitative Microscopy RNA Interference Regulation Research Ribosomes Risk Factors Role Site Staging Staining method Stains Sudan System Techniques Testing Therapeutic Tissues Transgenic Organisms Translations base biological adaptation to stress blastomere structure disease phenotype early onset feeding human disease in vivo infancy insight lipid metabolism loss of function meetings mutant nile red novel pH Homeostasis pre-doctoral promoter research study uptake

项目摘要

DESCRIPTION (provided by applicant): The intestinal Na+/H+ exchanger NHX-2 regulates both cellular pH and nutrient uptake in the nematode C. elegans, and loss-of-function leads to decreased fat stores and increased lifespan. In order to ask whether these phenotypes are mechanistically a result of altered pH or of caloric restriction, a global RNAi screen for other targets that alter intestinal cellular pH was performed. Almost a quarter of the 45 clones identified function in the mitochondria; in particular, ETC complex I was well-represented, as were proteins involved in mitochondria! ribosome function and in regulating mitochondrial morphology. Further testing suggested that several clones act at least in part cell non-autonomously. We propose here first to identify in which cells the non-autonomous functions occur using a novel system for cell specific RNAi. Then we will examine mitochondrial redox potential, pH, morphology, and energy production in vivo following RNAi in the relevant cells, as well as fat uptake and longevity in the targeted worms. Finally, we will target each clone in cultured nematode embryonic cells to directly test cell autonomy. The results of these experiments will provide mechanistic insights into how mitochondria influence intestinal cellular pH, as well as how this coupling influences fat metabolism and aging. Obesity has become a pressing health concern, particularly in younger people, over the last decade, and is one of the leading risk factors for developing diabetes and heart disease. Our research is aimed at defining how mitochondria, which meets the energy demands of the cell, influences pH homestasis in a well- conserved genetic model organism. Since cellular pH is linked to nutrient uptake, fat accumulation, and longevity, this functional coupling may represent a unique means of cellular communication and a novel target for anti-obesity therapeutics.

描述（由申请人提供）：肠道 Na+/H+ 交换剂 NHX-2 调节线虫的细胞 pH 值和营养吸收，功能丧失会导致脂肪储存减少和寿命延长。为了探究这些表型在机制上是否是 pH 值改变或热量限制的结果，对改变肠道细胞 pH 值的其他靶标进行了全局 RNAi 筛选。 45 个克隆中近四分之一被鉴定出在线粒体中发挥功能；特别是，ETC 复合物 I 以及参与线粒体的蛋白质都得到了很好的体现！核糖体功能和调节线粒体形态。进一步的测试表明，几个克隆至少部分地以细胞非自主方式行动。我们首先建议使用细胞特异性 RNAi 的新系统来识别哪些细胞中发生非自主功能。然后，我们将检查相关细胞中 RNAi 后的线粒体氧化还原电位、pH、形态和体内能量产生，以及目标线虫的脂肪摄取和寿命。最后，我们将针对培养的线虫胚胎细胞中的每个克隆来直接测试细胞自主性。这些实验的结果将为线粒体如何影响肠道细胞 pH 值，以及这种耦合如何影响脂肪代谢和衰老提供机制见解。在过去的十年中，肥胖已成为一个紧迫的健康问题，特别是对于年轻人来说，并且是患糖尿病和心脏病的主要危险因素之一。我们的研究旨在明确满足细胞能量需求的线粒体如何影响保守遗传模型生物体中的 pH 稳态。由于细胞 pH 值与营养吸收、脂肪积累和寿命有关，因此这种功能耦合可能代表了一种独特的细胞通讯方式和抗肥胖治疗的新靶点。