Minority Predoctoral Fellowship Program

少数族裔博士前奖学金计划

• 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
• 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

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+交换器NHX-2调节线虫C.秀隐杆线虫中的细胞pH和养分吸收，并且功能丧失导致脂肪存储下降和寿命增加。为了询问这些表型在机械上是pH值改变还是热量限制的结果，对改变了改变肠道细胞pH的其他靶标的全局RNAi屏幕。在45个克隆中，几乎四分之一在线粒体中识别出功能；特别是等等，我和线粒体涉及的蛋白质都非常有代表性！核糖体功能和调节线粒体形态。进一步的测试表明，几个克隆至少非自主作用。我们首先在这里提出，以确定使用用于细胞特异性RNAI的新型系统发生的非自主功能。然后，我们将检查相关细胞中RNAi后体内的线粒体氧化还原电位，pH，形态和能量产生，以及目标蠕虫中的脂肪吸收和寿命。最后，我们将靶向培养的线虫胚胎细胞中的每个克隆，以直接测试细胞自治。这些实验的结果将提供有关线粒体如何影响肠道细胞pH的机械见解，以及这种耦合如何影响脂肪代谢和衰老。 在过去的十年中，肥胖已成为紧迫的健康问题，尤其是在年轻人中，并且是发展糖尿病和心脏病的主要危险因素之一。我们的研究旨在定义符合细胞能量需求的线粒体如何影响保守的遗传模型生物体中的pH霍姆斯蒂。由于细胞pH与养分的摄取，脂肪的积累和寿命有关，因此该功能耦合可能代表了细胞通信的独特手段，也代表了抗肥胖治疗剂的新目标。