Role of GluN2A and MMPs in the CeA in Dependence-Induced Escalation of Etoh Drinking

CeA 中 GluN2A 和 MMP 在 Etoh 饮酒依赖性升级中的作用

基本信息

批准号：
10675691
负责人：
JOHN J. WOODWARD
金额：
$ 17.93万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
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：
2022-08-02 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10675691
关键词：
Acute Adolescent Adult Air Alcohol consumption Alcohol dependence Alcohol withdrawal syndrome Alcohols Amino Acids Amygdaloid structure Animals Anxiety Applications Grants Area Bilateral Brain CRISPR/Cas technology Calcium-Sensing Receptors Cannulas Cell Nucleus Chronic Clinical Research Data Dendritic Spines Dependence Effectiveness Electrophysiology (science)Ethanol Extracellular Matrix FDA approved Female Fiber Future Gelatinase B Gelatinases Genes Glutamates Guide RNA Health Heavy Drinking Human Implant Individual Infusion procedures Intoxication Ion Channel Gating Knock-in Mouse Knock-out Laboratories Learning Legal Status Ligands Light Link Matrix Metalloproteinase Inhibitor Matrix Metalloproteinases Measures Mediating Mediator Memory Molecular Mus N-Methyl-D-Aspartate Receptors N-Methylaspartate NMDA receptor A1 Neurons Outcome Permeability Pharmaceutical Preparations Pharmacological Treatment Phenotype Photometry Play Population Proteins Resistance Rewards Role Signal Transduction Single Nucleotide Polymorphism Slice Synapses Synaptic plasticity Testing Time Transgenic Mice Transmembrane Domain Vertebral column Viral Virus Western Blotting Wild Type Mouse Withdrawal Work Zinc addiction alcohol abuse therapy alcohol exposure alcohol measurement alcohol sensitivity alcohol use disorder binge drinking cell type cohort drinking drinking behavior effective therapy endonuclease in vivo inhibitor knock-down learned behavior male negative affect neurotransmission new growth novel postoperative recovery preclinical study prevent recruit reduce symptoms social

Acute Adolescent Adult Air Alcohol consumption Alcohol dependence Alcohol withdrawal syndrome Alcohols Amino Acids Amygdaloid structure Animals Anxiety Applications Grants Area Bilateral Brain CRISPR/Cas technology Calcium-Sensing Receptors Cannulas Cell Nucleus Chronic Clinical Research Data Dendritic Spines Dependence Effectiveness Electrophysiology (science)Ethanol Extracellular Matrix FDA approved Female Fiber Future Gelatinase B Gelatinases Genes Glutamates Guide RNA Health Heavy Drinking Human Implant Individual Infusion procedures Intoxication Ion Channel Gating Knock-in Mouse Knock-out Laboratories Learning Legal Status Ligands Light Link Matrix Metalloproteinase Inhibitor Matrix Metalloproteinases Measures Mediating Mediator Memory Molecular Mus N-Methyl-D-Aspartate Receptors N-Methylaspartate NMDA receptor A1 Neurons Outcome Permeability Pharmaceutical Preparations Pharmacological Treatment Phenotype Photometry Play Population Proteins Resistance Rewards Role Signal Transduction Single Nucleotide Polymorphism Slice Synapses Synaptic plasticity Testing Time Transgenic Mice Transmembrane Domain Vertebral column Viral Virus Western Blotting Wild Type Mouse Withdrawal Work Zinc addiction alcohol abuse therapy alcohol exposure alcohol measurement alcohol sensitivity alcohol use disorder binge drinking cell type cohort drinking drinking behavior effective therapy endonuclease in vivo inhibitor knock-down learned behavior male negative affect neurotransmission new growth novel postoperative recovery preclinical study prevent recruit reduce symptoms social

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项目摘要

Project Summary The legal status and widespread use of alcohol among the US population is associated with adverse health outcomes in both adolescents and adults. Nearly 88% of the US population have used alcohol at least once during their lifetime and rates of binge drinking and heavy alcohol use continue to be of concern. Pharmacological treatments for alcohol abuse show limited effectiveness and only three medications are FDA approved for treating alcohol dependence. One factor that underlies this problem is a lack of understanding of the mechanisms that contribute to excessive drinking. Recent results from our previous studies show that knock-in mice expressing ethanol resistant GluN2A N-methyl-D-aspartate receptors (NMDARs) drink the same as wild-type mice under baseline conditions but, unlike their wild-type counterparts, fail to escalate their drinking following repeated cycles of chronic intermittent ethanol exposure. These findings suggest that GluN2A NMDARs play a key role for in the loss of control over drinking observed in alcohol dependent subjects. Importantly, a similar lack of escalated drinking is observed following administration of inhibitors of zinc-dependent matrix metalloproteinases (MMPs) either i.c.v. or directly into the central nucleus of the amygdala (CeA). MMPs are key mediators of NMDA-mediated forms of synaptic plasticity where they remodel the extracellular matrix to support new growth and enlargement of glutamatergic dendritic spines. In this proposal we test the overarching hypothesis that MMPs and ethanol-sensitive GluN2A NMDARs in the CeA are critically involved in the transition to heavy drinking that is a hallmark of alcohol dependence. In Aim 1, we use slice electrophysiology, in vivo zymography and western blot analysis to test the hypothesis that CIE-induced changes in CeA neural signaling and MMP activity are absent in the GluN2A knock-in mice. In Aim 2, we use Crispr expressing AAVs and guide RNAs targeting the GluN2A gene to test the hypothesis that the CIE-induced increases in drinking and MMP activity requires functional GluN2A NMDARs in the CeA. The findings from these novel studies will provide critical data to support a future R01 grant application focused on determining which CeA cell types drive the altered drinking phenotype of the GluN2A knock-in mice and whether their expression in other key addiction-related brain areas contribute to this effect.

项目摘要美国人口中的法律地位和广泛使用酒精与不利健康有关青少年和成人的结果。近88％的美国人口至少使用过一次酒精在他们的一生中，饮酒和大量饮酒的发生率仍然令人关注。药理酗酒治疗的有效性有限，只有三种药物获得了FDA的批准治疗酒精依赖。这个问题基础的一个因素是对机制缺乏了解这有助于过度饮酒。我们先前研究的最新结果表明，敲门小鼠表达耐乙醇的Glun2a N-甲基-D-天冬氨酸受体（NMDAR）与野生型相同在基线条件下的小鼠，但与野生型的小鼠不同，他们无法升级饮酒。慢性间歇性乙醇暴露的反复周期。这些发现表明glun2a nmdars播放在酒精依赖受试者中观察到的饮酒失去控制中的关键作用。重要的是，类似的缺乏在给予锌依赖性基质抑制剂后，观察到升级的饮酒金属蛋白酶（MMP）i.c.v.或直接进入杏仁核（CEA）的中央核。 MMP是关键 NMDA介导的突触可塑性形式的介体，它们重塑细胞外基质以支持谷氨酸能树突状刺的新生长和扩大。在此提案中，我们测试了总体 MMP和对CEA中的MMP和乙醇敏感的Glun2a NMDAR与过渡至关重要大量饮酒是酒精依赖的标志。在AIM 1中，我们在体内使用切片电生理学 Zymography和Western印迹分析以检验CIE诱导的CEA神经信号变化的假设 Glun2a敲入小鼠中不存在MMP活性。在AIM 2中，我们使用CRISPR表达AAV和指南靶向Glun2a基因的RNA测试了CIE诱导的饮酒和MMP增加的假设活动需要CEA中的功能性Glun2a NMDAR。这些新研究的发现将提供关键支持未来R01赠款应用程序的数据，重点是确定哪种CEA单元类型驱动已更改 glun2a敲入小鼠的饮用表型，以及它们在其他关键成瘾有关的表达大脑区域有助于这种影响。