GluD1 regulation of structural plasticity in chronic ethanol exposure and protracted withdrawal

GluD1 对慢性乙醇暴露和长期戒断中结构可塑性的调节

• 批准号: 10724599
• 负责人: Daniel Tommis Christian
• 金额: $ 15.2万
• 依托单位: DES MOINES UNIV OSTEOPATHIC MEDICAL CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10724599
• 关键词: AMPA Receptors; Acute; Alcohol consumption; Alcohol dependence; Alcohol withdrawal syndrome; Amygdaloid structure; Automobile Driving; Behavioral; Biotinylation; Brain region; Cell physiology; Cells; Central Nervous System; Chronic; Confocal Microscopy; Data; Dendritic Spines; Development; Drug Exposure; Drug usage; Dyes; Electrophysiology (science); Ethanol; Female; Functional disorder; Future; Glutamate Receptor; Glutamates; Goals; Injections; Knock-out; Knowledge; Learning; Link; Maintenance; Measures; Mediating; Mediator; Membrane; Memory; Messenger RNA; Methods; Mission; Modeling; Morphology; N-Methyl-D-Aspartate Receptors; N-Methylaspartate; Neurobiology; Neurons; Output; Pathway interactions; Persons; Pharmaceutical Preparations; Play; Positioning Attribute; Predisposition; Procedures; Proteins; Rattus; Regulation; Relapse; Research; Role; Scaffolding Protein; Severities; Sprague-Dawley Rats; Substance Use Disorder; Substance abuse problem; Surface; Symptoms; Synapses; Synaptic Transmission; Synaptic plasticity; System; Testing; Time; Treatment outcome; United States National Institutes of Health; Vertebral column; Viral; Western Blotting; Withdrawal; Work; alcohol exposure; burden of illness; density; drug of abuse; drug withdrawal; experimental study; functional plasticity; knock-down; lucifer yellow; mRNA Expression; maladaptive behavior; male; new therapeutic target; novel; patch clamp; preference; receptor; receptor function; sex; small hairpin RNA; stimulant use; substance use treatment; synaptogenesis; targeted treatment; therapeutic development; therapeutic target; trafficking; treatment group; vapor

Project Summary Long-lasting vulnerability to relapse is a major hurdle to achieving successful treatment outcomes in persons with a substance use disorder. The glutamate receptor system is highly implicated in the development and maintenance of functional and structural alterations driving a return to drug use. Increasing our understanding of glutamatergic regulation of neurobiological alterations during drug use and withdrawal, we propose to explore a new target for the development of therapeutic agents. Delta-type ionotropic glutamate receptors (GluD) have been identified as regulators in the formation of synaptic connections, as they serve to regulate the alignment and development of dendritic spines. A recent study has indicated that GluD1 receptor function/expression plays a role in structural plasticity in a model of psychostimulant use. It is currently unknown what role GluD1 plays in altering structural plasticity following chronic ethanol exposure or protracted withdrawal. Using an ethanol exposure model this application seeks to identify the role GluD1 holds in the regulation of dendritic spine density and morphology under ethanol exposure and after protracted withdrawal. We will focus on the basolateral amygdala (BLA), a key region for ethanol action. We hypothesize that GluD1 dynamically controls chronic ethanol- and withdrawal-associated changes in spine morphology in the BLA. We will test this hypothesis in one Specific Aim, comprised of three independent experiments. All experiments will compare male and female rats. First, we will characterize dendritic spine morphology under control conditions, ethanol exposure, and across various withdrawal time points (day 1, 21, 42), using iontophoretic dye injection and confocal microscopy methods. We will also characterize GluD1 expression in surface membrane fractions using a biotinylation procedure followed by western blot analysis at each experimental time point. Finally, we will utilize dye injections and confocal microscopy paired with injection of a shRNA viral construct to knock down GluD1 levels in the BLA prior to ethanol exposure. This experiment will identify the specific role GluD1 plays in dendritic spine expression and morphology across our treatment groups and sex. Together, these studies will reveal the role of GluD1 in regulation of structural plasticity during ethanol exposure and withdrawal. As such, GluD1 regulation of structural plasticity could contribute to well characterized functional alterations of the glutamatergic receptor system during short term withdrawal. This would implicate GluD1 as a key hub, capable of regulating functional and structural plasticity, making it a promising target for therapeutic development. Long term objectives are to identify the overall impact of GluD1 regulation on both structural and functional plasticity across additional brain regions impacted by drug taking and withdrawal.

项目摘要 复发的长期脆弱性是在患者中实现成功治疗结果的主要障碍 患有药物滥用症谷氨酸受体系统高度参与了发育， 维持功能和结构改变，促使重新吸毒。增进我们的了解 药物使用和停药期间神经生物学改变的神经递质调节，我们建议 为治疗药物的开发探索新的靶点。δ型离子型谷氨酸受体 谷氨酸（GluD）已被确定为突触连接形成中的调节剂，因为它们用于调节 树突棘的排列和发育最近的研究表明，GluD1受体 功能/表达在精神兴奋剂使用模型中的结构可塑性中起作用。目前 尚不清楚GluD 1在慢性乙醇暴露或长期乙醇暴露后改变结构可塑性方面发挥什么作用 戒断使用乙醇暴露模型，本申请旨在确定GluD1在 调节树突棘密度和形态下乙醇暴露和长期戒断后。 我们将集中在基底外侧杏仁核（BLA），乙醇行动的关键区域。我们假设GluD1 动态控制BLA中脊柱形态的慢性乙醇和戒断相关变化。我们 我们将在一个特定目标中测试这一假设，该目标由三个独立的实验组成。所有实验将 比较雄性和雌性大鼠。首先，我们将在控制条件下表征树突棘形态， 乙醇暴露，并在不同的停药时间点（第1、21、42天），使用离子电渗染料注射 和共聚焦显微镜方法。我们还将表征表面膜组分中GluD1的表达， 使用生物素化程序，然后在每个实验时间点进行蛋白质印迹分析。最后我们 将利用染料注射和共聚焦显微镜与注射shRNA病毒构建体配对， 降低酒精暴露前BLA中GluD1的水平。本实验将确定GluD1的具体作用 在我们的治疗组和性别中的树突棘表达和形态学中发挥作用。所有这些 研究将揭示GluD1在乙醇暴露过程中调节结构可塑性的作用， 戒断因此，GluD1对结构可塑性的调节可能有助于充分表征功能性神经元的功能。 在短期戒断过程中的多巴胺能受体系统的改变。这将暗示GluD1是一种 关键枢纽，能够调节功能和结构可塑性，使其成为一个有前途的治疗目标 发展长期目标是确定GluD1调节对结构和功能的总体影响。 受药物服用和戒断影响的其他大脑区域的功能可塑性。