Cell Type Specific Control of Neuronal Signaling

神经元信号传导的细胞类型特异性控制

• 批准号: 8322176
• 负责人: Martilias Stephen Farrell
• 金额: $ 2.71万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2013-05-12
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8322176
• 关键词: ADORA2A gene; Acetylcholine; Adenosine; Affect; Animal Behavior; Basal Ganglia; Basal Ganglia Diseases; Behavior; Behavioral; Behavioral Sciences; Bioavailable; Brain; Brain region; Clozapine; Corpus striatum structure; Coupled; Couples; Coupling; Cyclic AMP; DARPP; Designer Drugs; Development; Disease; Drug Delivery Systems; Event; FOS gene; Family; Functional disorder; G Protein-Coupled Receptor Signaling; G(q) Alpha; G-Protein Signaling Pathway; G-Protein-Coupled Receptors; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Genetic; Hand; Human G(i) Alpha Proteins; Huntington Disease; Investigation; Knowledge; Laboratories; Lead; Ligands; MAPK3 gene; Measures; Mediating; Mental disorders; Mus; National Institute of Mental Health; Neurons; Obsessive-Compulsive Disorder; Oxides; Parkinson Disease; Pattern; Pharmaceutical Preparations; Phosphorylation; Physiological; Population; Receptor Activation; Research; Rewards; Scientist; Signal Pathway; Signal Transduction; Specificity; Technology; Testing; Therapeutic; Transgenic Mice; Transgenic Organisms; Work; addiction; cell type; design; in vivo; innovation; insight; neuronal circuitry; neuropsychiatry; promoter; receptor; receptor-mediated signaling; response; small molecule; spatiotemporal; tool

6. PROJECT SUMMARY / ABSTRACT Dysfunctional neuronal circuits are thought to be responsible for neuropsychiatric disorders, including obsessive-compulsive disorder, Parkinson¿s disease and addiction. G protein-coupled receptors (GPCRs) differentially modulate the activity of neuronal circuits by interacting with different G proteins, (e.g. G-alpha-s, G-alpha-i, and G-alpha-q-families), and these receptors are well-established drug targets. Distinct subpopulations of neurons comprising these circuits often express similar receptor subtypes, making it impossible to create drugs that selectively target receptors in specific circuits. In order to determine how receptors might modulate neuronal circuits, it is necessary to selectively activate GPCR signaling in a specific neuronal population in vivo. For the past two decades, efforts have been made to create tools to selectively manipulate GPCR signaling in distinct neuron populations - from small molecule ligands to genetic deletion - but these approaches have off-target effects. These off-target effects effectively eliminate the possibility to determine how GPCR signaling in a specific neuronal population affects circuitry function. A promising advancement to enable this specific manipulation is the development of the Designer Receptors Exclusively Activated by Designer Drug (DREADD) family of receptors. The objective of this proposal is to validate this recently created tool in vivo using a transgenic mouse expressing the G-alpha-s DREADD in a specific population of neurons in the striatum. Specific Aim 1 will validate the tool by determining whether the G-alpha-s DREADD activates canonical G-alpha-s signaling pathways by measuring cAMP accumulation, ERK1/2 phosphorylation, DARPP-32 phosphorylation, and c-Fos expression. Specific Aim 2 will validate the physiological relevance of the G-alpha-s DREADD-induced signaling by assessing its effects on animal behavior. The completion of this proposal will provide neuroscientists a powerful new tool to exclusively modulate GPCR signaling in specific neuronal populations.

6. 项目概要/摘要 功能失调的神经元回路被认为是导致神经精神疾病的原因，包括强迫症、帕金森病和成瘾。 G 蛋白偶联受体 (GPCR) 通过与不同的 G 蛋白（例如 G-alpha-s、G-alpha-i 和 G-alpha-q-家族）相互作用来差异调节神经元回路的活性，这些受体是公认的药物靶点。组成这些回路的不同神经元亚群通常表达相似的受体亚型，因此不可能制造出选择性靶向特定回路中受体的药物。为了确定受体如何调节神经元回路，有必要选择性激活体内特定神经元群中的 GPCR 信号传导。在过去的二十年里，人们一直在努力创造工具来选择性地操纵不同神经元群体中的 GPCR 信号传导——从小分子配体到基因删除——但这些方法具有脱靶效应。这些脱靶效应有效地消除了确定特定神经元群体中 GPCR 信号如何影响电路功能的可能性。实现这种特定操作的一个有前途的进步是设计药物独家激活的设计受体（DREADD）受体家族的开发。该提案的目的是使用在纹状体特定神经元群中表达 G-alpha-s DREADD 的转基因小鼠在体内验证这一最近创建的工具。具体目标 1 将通过测量 cAMP 积累、ERK1/2 磷酸化、DARPP-32 磷酸化和 c-Fos 表达来确定 G-alpha-s DREADD 是否激活经典 G-alpha-s 信号通路，从而验证该工具。具体目标 2 将通过评估 G-alpha-s DREADD 诱导的信号传导对动物行为的影响来验证其生理相关性。该提案的完成将为神经科学家提供一个强大的新工具来专门调节特定神经元群体中的 GPCR 信号传导。