High Content Functional Neuroanatomy of Endogenous GPCRs

内源性 GPCR 的高内涵功能神经解剖学

• 批准号: 10588941
• 负责人: Patrick Ross O'Neill
• 金额: $ 23.4万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-15 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10588941
• 关键词: Acceleration; Agonist; Anatomy; Animal Behavior; Antibodies; Autoradiography; Behavior; Binding; Biochemical; Biological Assay; Biosensor; Biotin; Brain; Brain Diseases; Chronic; Chronic stress; Color; Complex; Data; Development; Drug usage; Equipment; Exhibits; Fc Receptor; Fluorescence; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP-Binding Proteins; Genetic; Goals; Heating; Image; Immunohistochemistry; In Situ; Intervention; Ligands; Ligation; Link; Location; Measurement; Measures; Mediating; Metabotropic Glutamate Receptors; Methods; Modeling; Modification; Molecular; Motivation; Mus; National Institute of Drug Abuse; Neuroanatomy; Neurosciences; Nucleotides; Opioid Receptor; Pain; Pattern; Pharmaceutical Preparations; Pharmacological Treatment; Play; Property; Publishing; Rattus; Reagent; Resolution; Rewards; Role; Sensory; Signal Transduction; Specificity; Stress; Substance Use Disorder; Synaptic Transmission; System; Testing; Therapeutic; Tissue Fixation; Tissues; Withdrawal; addiction; anatomic imaging; biological adaptation to stress; cannabinoid receptor; cell type; chronic pain; cryostat; drug of abuse; drug seeking behavior; experimental study; fluorescence microscope; imaging modality; innovation; insight; interest; laboratory equipment; mu opioid receptors; negative affect; neural circuit; neuronal excitability; neurotransmitter release; nonhuman primate; novel; novel therapeutics; opioid use disorder; optogenetics; pharmacologic; protein activation; ratiometric; receptor; receptor coupling; receptor function; response; success; temporal measurement; tool; virtual

Abstract G protein coupled receptors (GPCRs) modulate neuronal excitability and neurotransmitter release, and exert profound effects on neural circuit functions. They play important roles in regulating motivation, reward, pain, negative affect, and stress responses, which are all important in the context of substance use disorders (SUDs). Exogenous ligands for several GPCRs have been shown to modify drug seeking behavior, which has generated interest in GPCRs as targets for the development of pharmacological therapeutics for substance use disorders. There are over 300 different non-sensory GPCRs expressed in mammalian brains, and fundamental questions remain unanswered for all of them. For example, how does a given receptor's ability to activate different G protein subtypes (Gi/o, Gs, Gq, G12/13) vary across neuroanatomical regions, cell types, and subcellular regions? How are these properties altered in response to chronic drug use, chronic stress, or chronic pain? To help answer these questions, we will develop new imaging methods in tissue sections for neuroanatomical measurement of GPCR-mediated G protein activation, with subcellular resolution and G protein subtype specificity. Our methods will only require common neuroscience equipment (cryostats and fluorescence microscopes) and commercially available reagents. They will generate high-content information on many GPCRs in healthy and diseased brains, and accelerate efforts to target GPCRs in addiction.

摘要 G蛋白偶联受体(GPCRs)调节神经元兴奋性和神经递质释放，并发挥作用 对神经回路功能产生深远影响。它们在调节动机、奖励、痛苦、 消极情绪和应激反应，这在物质使用障碍的背景下都是重要的 (肥皂水)。一些gpcr的外源配体已经被证明可以改变药物寻找行为，这已经 产生对GPCRs的兴趣，将其作为开发药物治疗药物的目标 使用障碍。哺乳动物的大脑中有300多种不同的非感官GPCR表达，并且 对于所有这些人来说，根本的问题仍然没有得到回答。例如，给定受体的能力是如何 激活不同的G蛋白亚型(Gi/o、Gs、Gq、G12/13)因神经解剖区域、细胞类型、 和亚细胞区域呢？这些特性是如何在慢性药物使用、慢性压力或 慢性疼痛?为了帮助回答这些问题，我们将在组织切片中开发新的成像方法 神经解剖学测量GPCR介导的G蛋白激活，亚细胞分辨率和G 蛋白质亚型特异性。我们的方法只需要普通的神经科学设备(低温恒温器和 荧光显微镜)和市场上可获得的试剂。它们将产生高内容的信息 在健康和患病的大脑中对许多GPCRs进行研究，并加快针对成瘾患者的GPCRs的努力。