Stable Isotope and Metabolomics Core

项目总结(见说明)： 稳定同位素和代谢组学核心的使命是与动物生理学核心合作，在复杂的体内代谢模型中提供有指导的代谢物和底物通量测定。稳定同位素和代谢组学核心提供了一系列体外代谢方法，增强了这些体内研究。这些服务为研究人员提供专门的分析，以确定细胞器、细胞、组织和全身水平的底物通量动态和代谢物分布，从而 阐明糖、蛋白质和脂类代谢紊乱的整合网络。通过与DRTC其他核心的合作努力，明确的药理、饮食、环境和遗传变化的影响被彻底表征为它们对葡萄糖稳态、胰岛素作用和新陈代谢的影响。候选分子在相关组织中的作用(即神经元， 通过在啮齿动物和其他模型中采用循序渐进的方法进行彻底和明确的体内和体外实验，可以具体描述与葡萄糖动态平衡相关的细胞(肝细胞、骨骼肌、脂肪细胞和β细胞)。为了实现这些目标，稳定同位素和代谢组学核心将：1)进行体内稳定同位素底物通量分析，以确定速率 蛋白质合成、脂肪生成、外周葡萄糖处理、肝脏葡萄糖循环、葡萄糖-甘油循环和葡萄糖-乳酸循环；2)使用SeaHorse Bioscience通量分析仪，在分离的细胞、组织外植体或组织培养中测定糖酵解(细胞外酸化率)和线粒体氧耗(线粒体呼吸)，以及更全面的稳定同位素通量评估；3)对血浆和组织代谢物的谱进行有针对性的假设驱动的评估 针对糖酵解/糖异生、磷酸戊糖和三羧酸(TCA)循环途径中的关键代谢物，以及脂肪代谢，包括脂肪酸、脂酰辅酶A和脂酰甘油胺的谱；以及4)在使用质谱仪的通量和代谢物图谱方法方面提供指导和协议制定，以评估与葡萄糖和脂肪酸稳态控制相关的分子生化指标。所有这些服务都提供给糖尿病研究的新手，以及从事糖尿病相关项目的研究人员，这些项目可以通过使用这一核心的专门知识和设施得到丰富和扩展。

PROJECT SUMMARY (See instructions): The mission of the Stable Isotope & Metabolomics Core is to provide guided metabolite and substrate flux determinations in complex in vivo metabolic models in partnership with the Animal Physiology Core. The Stable Isotope & Metabolomics Core provides an array of in vitro metabolic methodologies that augment these in vivo investigations. These services provide investigators with specialized assays to determine substrate flux dynamics and metabolite profiles at the organelle, cellular, tissue and whole body level thereby elucidating the integrative network of disorders in glucose, protein and lipid metabolism. Through these collaborative efforts with the other Cores of the DRTC, the effects of defined pharmacological, dietary, environmental and genetic alterations are thoroughly characterized for their effects on glucose homeostasis, insulin action, and metabolism. The role of candidate molecules in relevant tissues (i.e., neurons, hepatocytes, skeletal muscle, adipocytes and beta cells) that are related to glucose homeostasis can be specifically delineated by thorough and definitive in vivo and in vitro experimentation using a step-by-step guided approach in rodent, and other models. To accomplish these goals, the Stable Isotope & Metabolomics Core will: 1) perform in vivo stable isotope substrate flux assays for the determination of rates of protein synthesis, lipogenesis, peripheral glucose disposal, hepatic glucose recycling, glucose-glycerol cycling and glucose-lactate cycling; 2) detennine glycolysis (extracellular acidification rates) and mitochondrial oxygen consumption (mitochondrial respiration) in isolated cells, tissue explants or tissue culture, using Seahorse Biosciences Flux Analyzers, as well as more comprehensive stable isotope flux assessments; 3) perform targeted hypothesis driven assessments of plasma and tissue metabolite profiles for key metabolites in the glycolytic/gluconeogenic, pentose phosphate, and tricarboxylic (TCA) cycle pathways, and lipid metabolism, including fatty acid, fatty acyl CoA and fatty acyl camitine profiles; and 4) provide mentorship and protocol development in the use of mass spectrometer based flux and metabolite profiling methods for the evaluation of molecular biochemical targets relevant to the control of glucose and fatty acid homeostasis. All these services are available to investigators new to diabetes research, as well as to investigators working on diabetes-related projects that can be enriched and extended by the use of the expertise and facilities of this core.