Circulating endocannabinoids in rats: Assay development and validation

大鼠循环内源性大麻素：检测方法开发和验证

• 批准号: 9306814
• 负责人: Cecilia J Hillard
• 金额: $ 7.7万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9306814
• 关键词: 2-arachidonylglycerol; Acute; Adipose tissue; Adult; Affect; American; Americas; Animal Model; Animals; Behavior; Behavioral; Biological Assay; Blood; Blood Circulation; Brain; CNR1 gene; Calories; Cardiovascular Diseases; Cessation of life; Circadian Rhythms; Comorbidity; Consumption; Data; Daughter; Detection; Diabetes Mellitus; Drug Addiction; Eating; Eating Disorders; Endocannabinoids; Energy Intake; Epidemic; Ethanolamines; Exertion; Exhibits; Face; Fatty acid glycerol esters; Food; Food Energy; Food Intake Regulation; Food deprivation (experimental); Fright; Future; Glucose; Glycogen; Health; High Prevalence; Human; Hyperphagia; Individual; Intake; Ions; Isotopes; Lead; Life Expectancy; Ligands; Light; Lipids; Malignant Neoplasms; Measurement; Measures; Metabolic; Modernization; Mood Disorders; Muscle; Nutrient; Obesity; Odors; Paracrine Communication; Pattern; Peripheral; Physiological; Plasma; Play; Pre-Clinical Model; Process; Psychological Stress; Public Health; Rattus; Receptor Signaling; Regulation; Relapse; Research Project Grants; Rewards; Role; Scheme; Series; Signaling Molecule; Site; Societies; Source; Stress; System; Testing; Thermogenesis; Time; Tissues; Translational Research; Translations; United States; Validation; Vitamins; Withdrawal; acute stress; assay development; awake; brain circulation; design; emotional factor; experimental study; falls; food consumption; food restriction; human data; innovation; interest; liquid chromatography mass spectrometry; nutrition; response; trend; 2-arachidonylglycerol; Acute; Adipose tissue; Adult; Affect; American; Americas; Animal Model; Animals; Behavior; Behavioral; Biological Assay; Blood; Blood Circulation; Brain; CNR1 gene; Calories; Cardiovascular Diseases; Cessation of life; Circadian Rhythms; Comorbidity; Consumption; Data; Daughter; Detection; Diabetes Mellitus; Drug Addiction; Eating; Eating Disorders; Endocannabinoids; Energy Intake; Epidemic; Ethanolamines; Exertion; Exhibits; Face; Fatty acid glycerol esters; Food; Food Energy; Food Intake Regulation; Food deprivation (experimental); Fright; Future; Glucose; Glycogen; Health; High Prevalence; Human; Hyperphagia; Individual; Intake; Ions; Isotopes; Lead; Life Expectancy; Ligands; Light; Lipids; Malignant Neoplasms; Measurement; Measures; Metabolic; Modernization; Mood Disorders; Muscle; Nutrient; Obesity; Odors; Paracrine Communication; Pattern; Peripheral; Physiological; Plasma; Play; Pre-Clinical Model; Process; Psychological Stress; Public Health; Rattus; Receptor Signaling; Regulation; Relapse; Research Project Grants; Rewards; Role; Scheme; Series; Signaling Molecule; Site; Societies; Source; Stress; System; Testing; Thermogenesis; Time; Tissues; Translational Research; Translations; United States; Validation; Vitamins; Withdrawal; acute stress; assay development; awake; brain circulation; design; emotional factor; experimental study; falls; food consumption; food restriction; human data; innovation; interest; liquid chromatography mass spectrometry; nutrition; response; trend

The mechanisms that regulate the intake of food are vital for survival and health. While too little food consumption can lead to death, an equal or greater problem in 21st century America is the over-consumption of calorie-dense and inadequate consumption of vitamin-rich foods, resulting in obesity and poor nutrition. CB1 cannabinoid receptors (CB1R) in the brain play a vital role in the regulation of food intake, while activation of CB1R in peripheral tissues results in increased fat and glycogen storage; reduced glucose utilization by muscles; and reduced thermogenesis in brown adipose. Endocannabinoids are present in the human circulation and their concentrations are regulated by metabolic state and exhibit a circadian pattern. These previous findings lead us to the overall hypotheses that endocannabinoids in the circulation convey information about the need for additional caloric intake and that sustained, high endocannabinoid concentrations contribute to overeating. The objectives of this proposal are to establish and characterize an assay system in which endocannabinoid concentrations in the circulation of awake, freely behaving rats are determined. We will examine changes in endocannabinoid concentrations across the daily cycle; and following acute food restriction and stress, as these have all been shown to impact circulating endocannabinoid concentrations in humans. We will carry out three specific aims. The first aim is to optimize the endocannabinoid measurement in rat plasma; determining the limits of detection and quantification, and robustness. The second aim is to apply the assay to measure the circadian changes in circulating endocannabinoid concentrations in relationship to other metabolic signaling molecules; and to examine the effects of a brief period of food deprivation. The third aim will examine the time course of the effects of fear-evoking odor presentation on circulating endocannabinoids. Successful completion of these studies will provide the basis for future studies, including exploration of the mechanisms by which changes in energy status affect endocannabinoids in the circulation; the tissue source of the endocannabinoids in the circulation; and, most importantly, whether the endocannabinoids entering the brain from the circulation convey information about metabolic status to brain reward circuit activity. These studies are highly translatable to humans because endocannabinoid concentrations are easily measured in human blood. Indeed, the innovation of this project lies in its "beside to bench" translation of human observations to an animal model that is amenable to mechanistic studies. The studies in this project will increase our understanding of several processes important in the regulation of circulating endocannabinoid concentrations and will set the stage for future human and animal studies of their role in eating disorders, drug addiction and mood disorders.

调节食物摄入量的机制对于生存和健康至关重要。虽然食物太少 消费会导致死亡，在21世纪，美国的同等或更大的问题是过度消费 卡路里致密和富含维生素的食物的消费不足，导致肥胖和营养不良。 CB1 大脑中的大麻素受体（CB1R）在食物摄入调节中起着至关重要的作用，而激活 周围组织中的CB1R导致脂肪和糖原储存增加。减少葡萄糖利用 肌肉；并降低了棕色脂肪的热发生。内源性大麻素存在于人类中 循环及其浓度受代谢状态调节，并表现出昼夜节律模式。这些 先前的发现使我们提出了循环中内源性大麻素的总体假设 关于需要额外的热量摄入量以及持续的高内源性大麻素浓度有助于 暴饮暴食。该提案的目标是建立和表征一个测定系统 内源性大麻素浓度在清醒的循环中，确定自由行为的大鼠。我们将 检查整个周期内内源性大麻素浓度的变化；然后跟随急性食物 限制和压力，因为这些都被证明会影响循环的内源性大麻素浓度 人类。我们将执行三个具体目标。第一个目的是优化内源性大麻素的测量 在大鼠血浆中；确定检测和定量的限制以及鲁棒性。第二个目标是 应用测定法以测量循环内源性大麻素浓度的昼夜节律变化 与其他代谢信号分子的关系；并检查短暂食物的影响 剥夺。第三个目标将检查恐惧诱发的气味呈现对的时间过程 循环内源性大麻素。这些研究的成功完成将为以后的研究提供基础 包括探索能量状态变化影响内源性大麻素的机制 循环;循环中内源性大麻素的组织来源；而且，最重要的是，是否 内源性大麻素从循环中进入大脑，传达有关大脑代谢状态的信息 奖励电路活动。这些研究高度转化为人类，因为内源性大麻素 浓度很容易在人类血液中测量。确实，该项目的创新在于 替补席”将人类观察结果翻译成适合机械研究的动物模型。 该项目中的研究将增加我们对在调节中重要的几个过程的理解 循环内源性大麻素的浓度，并将为未来的人类和动物研究奠定基础 在饮食失调，吸毒和情绪障碍中的作用。