Phosphorylation and the CNS Actions of Ethanol

乙醇的磷酸化和中枢神经系统作用

• 批准号: 8054757
• 负责人: DORIT RON
• 金额: $ 35.71万
• 依托单位: ERNEST GALLO CLINIC AND RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8054757
• 关键词: Accounting; Acute; Address; Affect; Alcohol abuse; Alcohol consumption; Alcohol dependence; Alcohol withdrawal syndrome; Alcoholism; Area; Attenuated; Behavioral; Biochemical; Brain; Brain region; Corpus striatum structure; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Cytoplasmic Tail; Dependence; Development; Disease; Dorsal; Dose; Drug Delivery Systems; Enzymes; Ethanol; Event; Exposure to; FDA approved; Figs - dietary; Funding; Future; Gene Delivery; Glutamates; Goals; Habits; Health; Infusion procedures; Injection of therapeutic agent; Ion Channel; Lateral; Lead; Learning; Long-Term Potentiation; Measurement; Mediating; Mediator of activation protein; Memory; Methods; Modeling; Molecular; Molecular Conformation; Motivation; N-Methyl-D-Aspartate Receptors; NR1 gene; NR2B NMDA receptor; Obsessive compulsive behavior; Pathway interactions; Pharmaceutical Preparations; Phenotype; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Physiological; Play; Population; Post-Translational Protein Processing; Preparation; Process; Protein Tyrosine Phosphatase; Proto-Oncogene Proteins c-fyn; Rattus; Reagent; Relapse; Reporting; Research; Role; Self Administration; Signal Transduction; Site; Slice; Sucrose; Synaptic plasticity; Testing; Tyrosine; Tyrosine Kinase Inhibitor; Up-Regulation; Ventral Striatum; Viral; Withdrawal; alcohol effect; alcohol exposure; alcohol intervention; alcohol response; craving; drinking behavior; drug of abuse; ifenprodil; in vivo; inhibitor/antagonist; interest; nervous system disorder; neural circuit; novel; protein protein interaction; public health relevance; receptor; research study; src-Family Kinases

DESCRIPTION (provided by applicant): The long-term goal of our research is to test the hypothesis that post-translational modifications such as phosphorylation are major contributors to brain region-specific responses to alcohol exposure. In the first round of funding we found that the sensitivity of the N-methyl-D-Aspartate receptor (NMDAR) to ethanol in specific brain regions is determined by the activation state of the non-receptor tyrosine kinases Fyn (activation) and Src (inhibition), leading to changes in the phosphorylation state of the NR2B (Fyn) and NR2A (Src) subunits of the NMDAR. We found that these signaling alterations lead to profound changes in the activity of the NMDAR during and after acute exposure to ethanol in slice preparations and in vivo. More recently, we identified an important role for Fyn and NR2B-NMDAR in the dorsal striatum in the long-lasting facilitation of NR2B-containing NMDAR-mediated activity after ethanol exposure and in ethanol drinking behavior. The NMDAR is an important mediator of ethanol's actions in the brain, and our results suggest that changes in the activation state of the Fyn in the dorsal striatum contribute to aberrant synaptic plasticity that may underlie the development of behavioral phenotypes such as the propensity to consume ethanol. Using molecular, biochemical, electrophysiological and behavioral approaches, we propose to elucidate the mechanism leading to, and are the consequences of, ethanol-mediated Fyn activation in the dorsal striatum of rats. As Fyn activity is both positively and negatively regulated by phosphorylation, we will determine the contribution of the tyrosine phosphatases, PTPalpha and STEP, to ethanol's actions on Fyn activation, NR2B phosphorylation and long-term facilitation of NMDAR activity in vivo. Additionally, we plan to investigate possible physiological consequences of dorsal striatal activation of Fyn. Finally, we will determine the contribution of Fyn, PTPalpha, and STEP to operant ethanol self-administration in rats. Our long-term goal is to understand the molecular mechanisms that contribute to the development of disease states associated with alcohol addiction. Identification of new intracellular targets that are mediators of alcohol addiction is an essential future direction for the development of novel interventions for alcohol related disorders.7 PUBLIC HEALTH RELEVANCE: Alcoholism is a devastating disease that affects approximately 14 million people per year in the USA alone. Unfortunately, only limited numbers of drugs are currently approved by the FDA to treat adverse phenotypes associated with the disease. Therefore, there is a great need to identify novel targets for medication development. The ion channel NMDAR has attracted great interest as a potential drug target to treat alcoholism and other neurological diseases. Results generated from these studies may enable us to develop agents that modulate the activity of a subset population of the channel in specific areas of the brain, and therefore may lead to the development of novel, selective targets to treat alcohol abuse.

描述（由申请人提供）：我们的研究的长期目标是检验以下假设：诸如磷酸化之类的翻译后修饰是对酒精暴露的大脑区域特异性反应的主要因素。 In the first round of funding we found that the sensitivity of the N-methyl-D-Aspartate receptor (NMDAR) to ethanol in specific brain regions is determined by the activation state of the non-receptor tyrosine kinases Fyn (activation) and Src (inhibition), leading to changes in the phosphorylation state of the NR2B (Fyn) and NR2A (Src) subunits of the NMDAR。我们发现，这些信号改变会导致在切片制剂和体内急性暴露于乙醇中NMDAR的活性发生深刻的变化。最近，我们确定了FYN和NR2B-NMDAR在背侧纹状体中的重要作用，在乙醇暴露和乙醇饮用行为中含NR2B的NMDAR NMDAR介导的活性的长期促进中。 NMDAR是乙醇在大脑中作用的重要介体，我们的结果表明，背纹状体中Fyn的激活状态的变化有助于异常的突触可塑性，这可能是行为表型发展的基础，例如倾向，例如消除乙醇。使用分子，生化，电生理和行为方法，我们建议阐明导致乙醇介导的Fyn激活的机制，并且是大鼠背纹状体中Fyn激活的后果。由于FYN活性既受磷酸化的积极和负面调节，因此我们将确定酪氨酸磷酸酶，PTPALPHA和Step对乙醇对FYN激活，NR2B磷酸化的作用的贡献，以及对体内NMDAR活性的长期促进。此外，我们计划研究FYN背侧纹状体激活的可能生理后果。最后，我们将确定FYN，PTPALPHA的贡献，以及对大鼠操作乙醇自我给药的步骤。我们的长期目标是了解有助于与酒精成瘾有关的疾病状态发展的分子机制。鉴定新的细胞内靶标的是酒精成瘾的介体是开发新型酒精相关疾病干预措施的至关重要的未来方向。7 公共卫生相关性：酒精中毒是一种毁灭性的疾病，仅在美国就会影响约1400万人。不幸的是，FDA目前仅批准有限的药物来治疗与该疾病相关的不良表型。因此，迫切需要确定药物开发的新目标。 NMDAR的离子通道引起了人们对治疗酒精中毒和其他神经系统疾病的潜在药物的极大兴趣。这些研究产生的结果可能使我们能够开发用于调节大脑特定区域中该通道子集的活性的药物，因此可能导致发展新颖的选择性靶标以治疗酒精滥用。