The role of neurpopeptide Y in the arcuate nucleus of the hypothalamus in the regulation of energy expenditure and the involvment of its peripherally expressed receptor type-1 in the regulation of energy homeostasis in mice

下丘脑弓状核神经肽Y在能量消耗调节中的作用及其外周表达的1型受体在小鼠能量稳态调节中的作用

• 批准号: 231922378
• 负责人: Dr. Giacomo Mancini
• 金额: --
• 依托单位: Neurobiology Research Program
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2012
• 资助国家: 德国
• 起止时间: 2011-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/231922378?language=en
• 关键词: role; neurpopeptide; arcuate; nucleus; hypothalamus

Aim 1: Does NPY deletion in different neuron populations of the arc effect energy balance and cellular fuel selection?Body composition (whole body lean and fat mass), energy expenditure and physical activity in the conditional KO and control mice will be examined by indirect calorimetry at 9 weeks of age as well as at the end of the study at 16 weeks of age. Upon completion of the study plasma, brown adipose tissue (BAT) and white adipose tissue (WAT), liver and muscle will be dissected and the expression patterns/activity levels of various key regulators involved in mitochondrial function and thermogenesis in relevant tissues, including mitochondrial respiratory chain complexes (IV), citrate synthase, peroxisome proliferator-activated receptor ý co-activator (PGC-1a), uncoupling protein-1 (UCP-1) will be analyzed by real time PCR, Western blot analysis and/or enzyme activity assay. To verify whether differences in cellular fuel selection are present, we will analyze regulators involved in lipid oxidation such as carnitine palmitoyltransferase-1 (CPT-1), beta-hydroxyacyl CoA dehydrogenase (b-HAD) and medium-chain acyl-CoA dehydrogenase (MCAD) in several organs, such as liver, WAT, muscle. Furthermore, serum will be collected and the analysis of an extensive panel of hormones involved in the regulation of metabolism will be carried out. Does NPY deletion in different neuron populations of the arc affect energy expenditure via regulation of hypothalamic peptides?In order to see if any changes in energy expenditure can be attributed to changes in expression patterns and levels of hypothalamic peptides, brains from these mice will also be collected and mRNA expression levels of different peptides will be evaluated by using in situ hybridization. Particular emphasis will be given to the analysis of the mRNA expression levels in the Arc of POMC and CART, the opposing neurotransmitters to NPY and AGRP. Aim 2. To determine the contribution of peripheral Y1 receptors to the regulation of energy homeostasis.Does peripherally deletion of Y1 receptor effect energy balance and cellular fuel selection? By using a tamoxifen-inducuble mouse line, conditional Y1 knockout mice will be analyzed on SD conditions similarly to what described in Aim1.Does peripherally deletion of Y1 receptor protects against diet-induced obesity? In order to determine the effect of tissue specific Y1-receptor deletion in an obese context, a second set of mice will be analyzed in the same way but under high fat feeding conditions. The same parameters will be analyzed as described above. It is important to test these animals also under these conditions, as it will demonstrate which Y1 expressing tissue is most affected by high calorie and high fat intake.

目的1：神经肽Y在不同神经元中的缺失是否影响能量平衡和细胞燃料选择？在9周龄以及研究结束时16周龄，通过间接热量测定法检查条件性KO和对照小鼠的身体组成（全身瘦体重和脂肪量）、能量消耗和体力活动。研究完成后，将解剖血浆、棕色脂肪组织（BAT）和白色脂肪组织（WAT）、肝脏和肌肉，并检测相关组织中涉及线粒体功能和产热的各种关键调节因子的表达模式/活性水平，包括线粒体呼吸链复合物（IV）、柠檬酸合酶、过氧化物酶体增殖物激活受体β-辅激活因子（PGC-1 a）、解偶联蛋白-1（UCP-1）将通过真实的时间PCR、蛋白质印迹分析和/或酶活性测定进行分析。为了验证细胞燃料选择是否存在差异，我们将分析参与脂质氧化的调节剂，如肉毒碱棕榈酰转移酶-1（CPT-1），β-羟酰辅酶A脱氢酶（b-HAD）和中链酰基辅酶A脱氢酶（MCAD）在几个器官中，如肝脏，WAT，肌肉。此外，将采集血清，并对参与代谢调节的大量激素进行分析。神经肽Y缺失是否通过调节下丘脑肽影响弧的不同神经元群体的能量消耗？为了观察能量消耗的任何变化是否可归因于下丘脑肽的表达模式和水平的变化，还将收集这些小鼠的脑，并通过使用原位杂交来评价不同肽的mRNA表达水平。特别强调将给予POMC和CART的弧中的mRNA表达水平的分析，相反的神经递质，以NPY和AGRP。目标2.确定外周Y1受体对能量稳态调节的作用。外周Y1受体缺失是否影响能量平衡和细胞燃料选择？通过使用他莫昔芬诱导的小鼠系，条件性Y1基因敲除小鼠将在SD条件下进行分析，类似于Aim 1中所述。外周缺失Y1受体是否可以预防饮食诱导的肥胖？为了确定组织特异性Y1受体缺失在肥胖背景下的影响，将以相同的方式但在高脂肪饲养条件下分析第二组小鼠。将如上所述分析相同的参数。在这些条件下测试这些动物也很重要，因为它将证明哪种Y1表达组织受高热量和高脂肪摄入的影响最大。