The Effects of Chronic Manipulation of Proopiomelanocortin Neurons in Animal Models

长期操作阿黑皮素原神经元对动物模型的影响

• 批准号: 10425218
• 负责人: Caitlin Daimon
• 金额: $ 4.85万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10425218
• 关键词: Address; Affect; Animal Model; Animals; Anorexia; Behavioral Assay; Bilateral; Body Weight decreased; Bolus Infusion; Brain; Calcium; Canis familiaris; Chronic; Chronic Disease; Clozapine; Corticotropin; Couples; Cyclic AMP; Dependovirus; Diet; Disease; Doctor of Philosophy; Eating; Eating Disorders; Felis catus; Future; G-Protein-Coupled Receptors; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Goals; Health; Homeostasis; Hour; Human; Hypothalamic structure; Immunohistochemistry; Implant; Individual; Ligands; LoxP-flanked allele; Mediating; Messenger RNA; Mus; Muscarinics; Neurons; Neurotransmitters; Obesity; Oxides; Pathway interactions; Peptides; Persons; Pharmaceutical Preparations; Physiologic Monitoring; Population; Pro-Opiomelanocortin; Pump; Regulation; Research; Research Proposals; Rodent Model; Role; Science; Scientist; Severities; Structure of nucleus infundibularis hypothalami; Technology; Testing; Time; Training; Weight; base; career; desensitization; designer receptors exclusively activated by designer drugs; diet-induced obesity; doctoral student; energy balance; equilibration disorder; feeding; immunoreactivity; inhibitor; inward rectifier potassium channel; mortality risk; neurotransmission; novel therapeutics; obesity treatment; obesogenic; prevent; programs; receptor; reduced food intake; response; success; targeted treatment; therapy development

Project Summary This proposal seeks to train a dual degree, DVM-PhD, student to prepare her for success in a career as an independent clinician-scientist. The applicant will earn a PhD in Biomedical Sciences while simultaneously earning a DVM. The proposed research seeks to determine whether proopiomelanocortin (POMC) neurons in the hypothalamus are a viable target for potential therapies for disorders of energy balance regulation, including obesity and eating disorders. Given that 35-40% of people in the US are obese and 2-5% suffer from eating disorders, it is critically important that we better understand how the brain regulates energy homeostasis. The long-term objective of this proposal is to determine whether chronic manipulation of hypothalamic POMC neurons will correct disturbances in food intake and bodyweight in animal models of energy balance disorders. The following specific aims will be addressed: 1) Determine the impact of chronically stimulating POMC neurons in an animal model of obesity. 2) Determine the impact of chronically inhibiting POMC neurons in an animal model of anorexia. Chronic stimulation of POMC neurons in mice fed an obesogenic diet (Aim 1) and chronic inhibition of POMC neurons in mice undergoing the activity-based anorexia behavioral assay (Aim 2) will be achieved using chemogenetic (Designer Receptors Exclusively Activated by Designer Drugs (DREADD)) technology. Adeno-associated virus technology will be used to deliver either stimulatory hM3Dq (Aim 1) or inhibitory hM4Di (Aim 2) DREADDs specifically to POMC neurons in the hypothalamus. Clozapine-n-oxide, an inert ligand that only activates DREADDs, will be administered to mice in discrete boluses via implantable, programmable microinfusion pumps. In addition to monitoring physiological endpoints such as bodyweight and food intake, RNAscope will be used to determine the degree of POMC neuron activation or inhibition. Immunohistochemistry will be used to verify adequate DREADD expression in POMC neurons. Altogether, this research proposal will determine whether chronic manipulation of POMC neurons will correct the food intake and bodyweight disturbances observed in rodent models of obesity and anorexia. In addition, the results of this proposal will be of value for future studies aimed at developing therapies for energy balance disorders.

项目摘要 这项建议旨在培养一个双学位，DVM-PhD，学生为她在职业生涯中取得成功做好准备， 独立临床科学家申请人将获得生物医学科学博士学位，同时 获得DVM。这项拟议中的研究旨在确定是否阿黑皮素原（POMC）神经元在 下丘脑是能量平衡调节障碍的潜在治疗的可行靶点， 包括肥胖和饮食失调。鉴于美国35-40%的人肥胖，2-5%的人患有肥胖症 饮食失调，这是至关重要的，我们更好地了解大脑如何调节能量 体内平衡这项建议的长期目标是确定长期操纵 下丘脑POMC神经元将纠正动物模型中的食物摄入和体重紊乱， 能量平衡失调将讨论以下具体目标：1）确定长期 刺激肥胖动物模型中的POMC神经元。2)确定长期抑制 厌食症动物模型中的POMC神经元。慢性刺激小鼠的POMC神经元， 肥胖饮食（目标1）和慢性抑制POMC神经元的小鼠进行基于活动的 厌食行为测定（目标2）将使用化学遗传学（Designer Receptors Exclusively 由Designer Drugs（DREADD）技术激活。腺相关病毒技术将用于 将刺激性hM 3Dq（Aim 1）或抑制性hM 4Di（Aim 2）DREADD特异性递送至POMC神经元 在下丘脑氯氮平-n-氧化物，一种仅激活DREADD的惰性配体，将被施用给 小鼠通过可植入的可编程微输注泵进行离散推注。除了监测 生理终点，如体重和食物摄入量，RNAscope将用于确定程度 POMC神经元的激活或抑制。将使用免疫组织化学验证DREADD是否充分 POMC神经元表达。总之，这项研究建议将确定慢性操纵是否 的POMC神经元将纠正在啮齿动物模型中观察到的食物摄入和体重紊乱， 肥胖和厌食症。此外，这一建议的结果将对今后的研究具有价值， 开发能量平衡失调的疗法