Pathway-specific Fyn signaling in the striatum and ethanol drinking

纹状体和乙醇饮用中的通路特异性 Fyn 信号传导

• 批准号: 8930906
• 负责人: DORIT RON
• 金额: $ 34.59万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-25 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8930906
• 关键词: AMPA Receptors; Address; Agonist; Alcohol abuse; Alcohol consumption; Behavior; Behavioral; Binding; Brain region; Corpus striatum structure; Coupled; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; DRD2 gene; Data; Designer Drugs; Development; Disease; Dopamine; Dopamine D1 Receptor; Dopamine D2 Receptor; Ethanol; Event; Exhibits; Extramural Activities; Fluorescence; GTP-Binding Proteins; Gene Delivery; Genes; Genetic; Genetically Engineered Mouse; Goals; Health; Laboratories; Light; Maintenance; Mediating; Methodology; Methods; Modeling; Molecular; Mus; N-Methyl-D-Aspartate Receptors; Neurons; Neurosciences; Outcome; Pathway interactions; Pharmaceutical Preparations; Phosphotransferases; Physiological; Physiological Adaptation; Play; Proto-Oncogene Proteins c-fyn; Resolution; Rewards; Rodent; Role; Self Administration; Signal Pathway; Signal Transduction; Synapses; Synaptic Membranes; System; Techniques; Testing; Transgenic Mice; United States National Institutes of Health; Viral; addiction; alcohol response; alcohol seeking behavior; base; cell type; drinking; drinking behavior; genetic approach; in vivo; innovation; learned behavior; motivated behavior; neuroadaptation; novel; receptor; recombinase; tool

DESCRIPTION (provided by applicant): This proposal is aimed at studying signaling-dependent neuroadaptations that occur in response to ethanol in subpopulations of neurons within the dorsomedial striatum (DMS). The DMS plays a central role in goal- directed behaviors and addiction, and we found that voluntary and passive exposure of rodents to ethanol activates the tyrosine kinase Fyn specifically in the DMS. We further discovered that in response to ethanol, activated Fyn phosphorylates the NR2B subunit of the NMDARs (GluN2B) resulting in increased synaptic activity of NMDA and AMPA receptors (NMDAR/AMPAR). Finally, we showed that the Fyn-signaling pathway in the DMS plays a crucial role in mechanisms underlying ethanol-drinking behaviors. Neurons in the DMS express either dopamine (DA) D1 or D2 receptors (D1R, D2R) that form the direct "Go" and indirect "No-Go" pathways. Thus, DA is thought to have different and possibly opposite effects on D1R and D2R neurons. D1Rs are coupled to G protein �s (Gs), which activates cAMP/PKA signaling, whereas D2Rs are coupled to Gi, which inhibits cAMP/PKA. We found that Fyn activation depends on cAMP/PKA, and that in vivo administration of a D1R agonist but not D2R agonist activates Fyn signaling in the DMS, and specifically in D1R DMS neurons. Together, these data suggest the DA activates a signaling pathway consisting of D1R/Gs/Fyn/GluN2B/AMPAR in D1R neurons. Ethanol increases DA levels in the striatum, and we hypothesize that ethanol activates Fyn signaling specifically in D1R neurons in the DMS to facilitate neuroadaptations in the direct "Go" pathway that drive ethanol drinking behaviors. We also hypothesize that remote activation of Gs signaling in D1R DMS neurons is sufficient to produce similar Fyn-dependent cellular and behavioral neuroadaptations. These hypotheses will be tested in vivo using transgenic mice that express Cre in D1R or D2R neurons, and will utilize innovative molecular and chemico- genetic approaches to manipulate single genes (Cre-FLEX), and to remotely activate Gs signaling using the Designer Receptors Exclusively Activated by Designer Drugs (DREADD) method in specific subpopulations of neurons. Aim 1 and Aim 2 will determine whether Fyn-dependent adaptations induced by ethanol occur in D1R, but not D2R, neurons in the DMS, and whether Gs DREADD activation in DMS D1R neurons is sufficient to produce similar outcomes. Aim 3 will investigate the contribution of Fyn signaling in D1R DMS neurons to ethanol drinking behaviors, and whether Gs DREADD activation of Fyn in D1R is sufficient to produce ethanol seeking and drinking. Combining Molecular and Systems Neuroscience methodologies, we will unravel cell- type specific signaling adaptations that underlie the development and maintenance of ethanol drinking and seeking behaviors.

描述（由适用提供）：该提案旨在研究信号依赖性神经适应于对乙醇在背侧纹状体（DMS）内神经元亚群中发生的响应。 DMS在目标定向的行为和成瘾中起着核心作用，我们发现啮齿动物对乙醇的自愿和被动暴露会在DMS中专门激活酪氨酸激酶FYN。我们进一步发现，激活的Fyn响应于乙醇，磷酸化NMDARS（GLUN2B）的NR2B亚基，从而导致NMDA和AMPA受体（NMDAR/AMPAR）的突触活动增加。最后，我们表明，DMS中的FYN信号途径在乙醇饮用行为的基础机制中起着至关重要的作用。 DMS中的神经元表达形成直接“ go”和间接“ no-go”途径的多巴胺（DA）D1或D2受体（D1R，D2R）。这就是DA对D1R和D2R神经元具有不同的和可能的相反作用。 D1R与G蛋白（GS）耦合，该蛋白（GS）激活CAMP/PKA信号，而D2RS与GI耦合，GI抑制CAMP/PKA。我们发现FYN激活取决于CAMP/PKA，并且在体内给药D1R激动剂，但不为D2R激动剂激活DMS中的FYN信号，特别是在D1R DMS神经元中。总之，这些数据表明DA激活了由D1R神经元中D1R/GS/FYN/GLUN2B/AMPAR组成的信号通路。乙醇增加了纹状体中的DA水平，我们假设乙醇在DMS的D1R神经元中专门激活FYN信号传导，以促进直接“ GO”途径中的神经照顾，从而驱动乙醇饮用行为。我们还假设D1R DMS神经元中GS信号传导的远程激活足以产生类似的FYN依赖性细胞和行为神经适应。 These hypotheses will be tested in vivo using transgenic mice that express Cre in D1R or D2R neurons, and will utilize innovative molecular and chemico- genetic approaches to manipulate single genes (Cre-FLEX), and to remotely activate Gs signaling using the Designer Receptors Exclusively Activated by Designer Drugs (DREADD) method in specific subpopulations of neurons. AIM 1和AIM 2将确定乙醇在D1R中诱导的FYN依赖性适应性是否发生在D1R中，而不是DMS中的D2R，以及DMS D1R神经元中的GS Dreadd激活是否足以产生类似的结果。 AIM 3将研究D1R DMS神经元中FYN信号传导对乙醇饮用行为的贡献，以及D1R中FYN的GS Dreadd激活是否足以产生寻求乙醇的寻求和饮用。结合了分子和系统神经科学方法，我们将揭示细胞类型的特定信号传导适应，以发展和维持乙醇饮酒和寻求行为。