Gao signaling mechanisms in the striatum

纹状体中的Gao信号传导机制

• 批准号: 10665127
• 负责人: Cody William Kowalski
• 金额: $ 1.65万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-16 至 2022-12-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10665127
• 关键词: Acute; Address; Adenylate Cyclase; Analgesics; Behavior; Behavioral; Biosensor; Brain; Chronic; Clinical; Complex; Corpus striatum structure; Coupling; Cre driver; Cyclic AMP; DRD2 gene; Dependence; Disinhibition; Dopamine; Dopamine Receptor; Electrophysiology (science); Epidemic; Equilibrium; Euphoria; G-Protein-Coupled Receptors; GTP-Binding Proteins; Glutamates; Goals; Guanosine Triphosphate; Heterotrimeric GTP-Binding Proteins; Image; Impairment; Knock-out; Label; Learning; Measures; Mediating; Morphine; N-Methylaspartate; Neurons; Nucleus Accumbens; Opiate Addiction; Opioid; Opioid Analgesics; Optics; Overdose; Pathway interactions; Phenotype; Physiology; Positive Reinforcements; Process; Production; Psychological reinforcement; RGS Proteins; Receptor Activation; Receptor Signaling; Research; Resolution; Rewards; Role; Second Messenger Systems; Self Administration; Signal Pathway; Signal Transduction; Signaling Protein; Slice; Source; Specificity; Structure; Substance Use Disorder; Synapses; Synaptic plasticity; Testing; Therapeutic; Time; Transducers; Ventral Tegmental Area; Withdrawal; addiction; clinical application; effective therapy; experimental study; in vivo; knock-down; mesolimbic system; mu opioid receptors; neuroregulation; new therapeutic target; opiate tolerance; opioid abuse; opioid epidemic; opioid use; optogenetics; postsynaptic; receptor coupling; receptor expression; response; reward circuitry; side effect; signal processing; treatment strategy; two-photon

Project Summary/Abstract Opioid side effects including euphoria, tolerance and dependence limit clinical applications and have increasingly contributed to fatal misuse. Opioid analgesia, reinforcement, and withdrawal are mediated by the μ-opioid receptor, that is expressed throughout the reward circuit of the brain. Our long-term goal is to identify mechanisms underlying and regulating μ-opioid receptor coupling to downstream pathways, and how these respective pathways contribute to cellular and synaptic plasticity that promote addiction and dependence. In addition to its acute neuromodulatory effects, cyclic AMP (cAMP) signaling is intimately involved in modulating cellular, structural and synaptic plasticity from repeated opioid administration. Unlike the μ-opioid effectors Gαi and Gβγ, our understanding of the mechanistic role of Gαo in transducing MOR signals is limited. Our lab and others have supported a role of Gαo in coupling μ-opioid receptor activation to cAMP signaling using knockdown approaches or by manipulating specific Regulators of G-Protein Signaling proteins. More recently, we have identified changes in cAMP levels, opioid reinforcement behavior, and withdrawal phenotypes resulting from striatal conditional Gαo knockout. We propose that these effects of Gαo deletion result from impaired MOR coupling to AC, via modulation of Gβγ availability. In Aim 1, I will determine the contribution of Gαo to opioid- evoked cAMP response dynamics of cultured striatal neurons using a genetically encoded optical cAMP biosensor pioneered by our lab. This approach will be further applied in combination with optogenetic stimulation of striatal inputs in brain slices to investigate how Gαo contributes to opioid induced adaptations of striatal dopamine signal integration in an intact mesolimbic circuit. In Aim 2, I will use electrophysiology to determine how Gαo contributes to opioid-induced adaptations that regulate striatal physiology and synaptic plasticity. In Aim 3, I will determine the specific striatal circuits in which Gαo influences opioid reinforcement behaviors. These studies will identify mechanistic contributions of Gαo to μ-opioid receptor signaling, subsequent adaptations that modify striatal physiology, and how these effects ultimately influence opioid reinforcement behavior within discrete striatal circuits. This research has the potential to identify new strategies for manipulating opioid reinforcement for therapeutic applications, to address the ongoing epidemic of opioid abuse.

项目摘要/摘要 阿片类药物的副作用，包括欣快，耐受性和依赖性限制临床应用，并增加了 导致致命滥用。阿片类镇痛，增强和戒断由μ阿片类药物介导 接收器，在整个大脑的奖励电路中表达。我们的长期目标是确定 基础和调节μ-阿片受体耦合到下游途径的机制，以及如何 各自的途径有助于促进成瘾和依赖性的细胞和突触可塑性。在 除了其急性神经调节作用，环状AMP（CAMP）信号与调节密切相关 反复给药的细胞，结构和突触可塑性。与μ阿片类药物效应GαI不同 和Gβγ，我们对GαO在传输MOR信号中机械作用的理解是有限的。我们的实验室和 其他人则支持GαO在使用敲低的cAMP传导耦合μ-阿片受体激活中的作用 方法或通过操纵G蛋白信号蛋白的特定调节剂。最近，我们有 确定了营地水平的变化，阿片类药物增强行为和戒断表型 纹状体条件GαO敲除。我们提出，GαO缺失的这些影响是由于MOR受损而导致的 通过调节Gβγ的可用性，耦合到AC。在AIM 1中，我将确定GαO对蛋白质的贡献 使用一般编码的光学营的培养纹状体神经元的诱发营地反应动力学 生物传感器由我们的实验室开创。该方法将与光遗传刺激相结合进一步应用 脑切片中的纹状体输入，以研究GαO如何促进OID诱导的纹状体适应 在完整的中边路电路中的多巴胺信号整合。在AIM 2中，我将使用电生理学来确定 GαO如何有助于阿片类药物诱导的适应，以调节纹状体生理和突触可塑性。在 AIM 3，我将确定GαO影响Ooid增强行为的特定纹状体回路。这些 研究将确定GαO对μ阿片受体信号传导的机理贡献，随后的适应 修改纹状体生理学，以及这些影响如何最终影响阿片类药物的增强行为 离散的纹状体电路。这项研究有可能确定操纵Ooid的新策略 用于治疗应用的强化，以解决阿片类药物滥用的持续流行。