Hypothalamic Integration of Energy Homeostasis Signals

下丘脑能量稳态信号整合

基本信息

批准号：
7475873
负责人：
Gregory Stefan Barsh
金额：
$ 31.08万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-01 至 2009-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7475873
关键词：
ART protein Acute Acyl Coenzyme A Affect Alleles Amphetamines Animals Anorexia Attention Bacterial Artificial Chromosomes Bacterial Chromosomes Behavior Behavioral Binding Biological Assay Blood Circulation Body Composition Brain Calorimetry Cell membrane Cells Cocaine Cultured Cells Cytoplasm Elevation Energy Metabolism Engineering Environment Escherichia coli Exposure to Feeding behaviors Food deprivation (experimental)Gene Expression Gene Targeting Genes Genetic Crosses Growth Homeostasis Hormonal Hormones Hour Hypothalamic structure Image Individual Injection of therapeutic agent Insulin Insulin Receptor Insulin Signaling Pathway Knock-out Leptin Leptin resistance Measurement Measures Mediating Membrane Messenger RNA Metabolic Metabolism Mitochondrial Carnitine Palmitoyltransferase Pathway Modeling Molecular Mus Neurons Neuropeptides Nutrient Obesity Oleic Acid Oleic Acids PH Domain Pathway interactions Pattern Peripheral Phenotype Phosphatidylinositols Phosphorylation Phosphotransferases Play Preparation Proteins Purpose Receptor Activation Receptor Gene Regulatory Element Reporter Research Personnel Resolution Role Signal Transduction Site Slice Stimulus Stomach Structure of nucleus infundibularis hypothalami Technology Time Tissues Transcript Transferase Transgenic Organisms Work base cell type day design energy balance extracellular feeding ghrelin homologous recombination inhibitor/antagonist insight insulin signaling leptin receptor molecular imaging mutant neuropeptide Y neurotransmission platelet protein P47 pre-pro-opiocortin prevent programs recombinase research study response tool development

项目摘要

Homeostatic control of adiposity depends on the ability of the brain to sense and respond to changes in circulating indicators of body energy stores. This proposal is focused on understanding the molecular and cellular mechanisms by which two groups of neurons in the arcuate nucleus of the hypothalamus, those marked by expression of Agouti-related protein (Agrp) and Preproopiomelanocortin (Pomc), measure and respond to changes in leptin, insulin, and ghrelin. Our approach has been made possible by recent advances in transgenic technology and molecular imaging, and is based on the development of tools that allow us to control gene expression specifically in the Agrp/Npy and Pomc/Cart neurons. Using homologous recombination of bacterial artificial chromosome (BAC) clones in E. coli followed by transgenic introduction of the modified clones into mice, we have placed Cre recombinase under control of regulatory elements for the Agrp or Pomc genes. These Tg.AgrpCre and Tg.PomcCre mice will be applied to better understand how peripheral signals of body fuel stores and meal patterning are perceived and integrated by hypothalamic neurons by constructing and analyzing the phenotype of mice that are deficient for Stat3 or for Irs2 in either Agrp neurons, Pomc neurons, or both. We will also use a newly developed assay for dynamic measurement of phosphoinositide 3-kinase signaling in individual cells to determine how each subpopulation of arcuate nucleus neurons responds to leptin, insulin, or ghrelin in a brain slice preparation. Finally, we will determine if and how agents that increase long chain fatty acyl CoA levels intersect with Stag and PI3K signaling in Agrp and Pomc neurons. Together, these studies will provide fundamental new insight into cellular mechanisms governing the response of key hypothalamic neurons to hormonal and nutrient-related signals.

肥胖的稳态控制取决于大脑感知和响应身体能量存储指标的变化的能力。该建议的重点是理解分子和细胞机制，通过这种机制，下丘脑的两组神经元在下丘脑的核核中，这些神经元以Agouti相关的表达为标志。蛋白质（AGRP）和前甲状腺皮质素（POMC），测量和响应瘦素，胰岛素和生长素蛋白的变化。转基因技术和分子成像的最新进展使我们的方法成为可能，并且基于工具的开发，使我们能够在AGRP/NPY和POMC/CART神经元中专门控制基因表达。使用大肠杆菌中细菌性人造染色体（BAC）克隆的同源重组，然后转基因将改良的克隆引入小鼠中，我们将CRE重组酶置于AGRP或POMC基因的调节元件下。这些TG.AGRPCRE和TG.POMCCRE小鼠将被应用以更好地了解外围下丘脑神经元通过构建和分析AGRP神经元，POMC神经元或两者兼有IRS2的小鼠的表型来感知和整合身体燃料存储和粉状模式的信号。我们还将使用新开发的测定法进行动态测量单个细胞中磷酸肌醇3-激酶信号的动态测量，以确定弧形神经元的每个亚群如何响应瘦素，胰岛素或生长素蛋白在大脑切片准备中。最后，我们将确定在AGRP和POMC神经元中增加长链脂肪酰基COA水平与雄鹿和PI3K信号的相交的药物是如何以及如何相交的。总之，这些研究将提供对负责下丘脑神经元对激素和营养相关信号的反应的细胞机制的基本新见解。