Gao signaling mechanisms in the striatum

纹状体中的Gao信号传导机制

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要阿片类药物的副作用，包括欣快感、耐受性和依赖性，限制了临床应用，并且越来越多导致致命的阿片类药物镇痛、强化和戒断是由 μ-阿片类药物介导的。受体，在整个大脑的奖励回路中表达，我们的长期目标是识别。 μ-阿片受体与下游途径偶联的潜在机制和调节机制，以及这些机制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。除了其急性神经调节作用外，环磷酸腺苷 (cAMP) 信号传导还密切参与调节与 μ-阿片效应器 Gαi 不同，重复施用阿片类药物可产生细胞、结构和突触可塑性。和 Gβγ，我们对 Gαo 在 MOR 信号转导中的机制作用的理解是有限的。其他人支持 Gαo 通过敲低将 μ-阿片受体激活与 cAMP 信号传导耦合的作用最近，我们有了一些方法或通过操纵 G 蛋白信号蛋白的特定调节因子。确定了 cAMP 水平、阿片类药物强化行为和戒断表型的变化我们认为，纹状体条件性 Gαo 敲除是由于 MOR 受损造成的。通过调节 Gβγ 可用性与 AC 耦合。在目标 1 中，我将确定 Gαo 对阿片类药物的贡献。使用基因编码的光学 cAMP 诱发培养纹状体神经元的 cAMP 反应动力学我们实验室首创的生物传感器将与光遗传学刺激结合进一步应用。脑切片中的纹状体输入，以研究 Gαo 如何促进阿片类药物诱导的纹状体适应在目标 2 中，我将使用电生理学来确定完整的中脑边缘回路中的多巴胺信号整合。 Gαo 如何促进阿片类药物诱导的调节纹状体生理学和突触可塑性的适应。目标 3，我将确定 Gαo 影响阿片类药物强化行为的特定纹状体回路。研究将确定 Gαo 对 μ-阿片受体信号传导的机制贡献，以及随后的适应改变纹状体生理学，以及这些效应最终如何影响纹状体内部的阿片类药物强化行为这项研究有可能确定操纵阿片类药物的新策略。加强治疗应用，以解决持续流行的阿片类药物滥用问题。