Pathological AMPA receptor adaptations governing dependence-escalated alcohol self-administration

病理性 AMPA 受体适应控制依赖性升级的酒精自我给药

• 批准号: 10592002
• 负责人: Jessica Lea Hoffman
• 金额: $ 13.06万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10592002
• 关键词: AMPA Receptors; Address; Alcohol abuse; Alcohol consumption; Alcohol dependence; Alcohols; Amygdaloid structure; Automobile Driving; Behavior; Behavior assessment; Behavioral; Binding; Brain; Brain region; Calcium; Calcium Signaling; Chronic; Confocal Microscopy; Data; Dependence; Development; Ethanol; Fiber; Gene Expression; Genetic; Glutamates; Hippocampus; Infusion procedures; Integral Membrane Protein; Knowledge; Learning; Link; Maintenance; Measures; Medial; Mediating; Membrane; Memory; Methods; Modeling; Molecular; Mus; Nucleus Accumbens; Pathologic; Pathology; Pathway interactions; Permeability; Pharmacology; Phase; Phosphorylation; Photometry; Prefrontal Cortex; Process; Property; Prosencephalon; Proteins; Receptor Inhibition; Regulation; Resistance; Rewards; Role; Scientist; Self Administration; Severities; Site; Synaptic Transmission; Synaptic plasticity; Techniques; Testing; Training; Up-Regulation; Withdrawal; Work; alcohol abuse therapy; alcohol effect; alcohol exposure; alcohol reinforcement; alcohol response; alcohol use disorder; behavioral plasticity; brain reward regions; calmodulin-dependent protein kinase II; career; craving; drinking; excitatory neuron; experience; experimental study; genetic regulatory protein; inhibitor; innovation; insight; interdisciplinary approach; multidisciplinary; neural; neuromechanism; novel; pharmacologic; pre-clinical; preclinical study; receptor expression; receptor function; selective expression; trafficking; transmission process; vapor

Project Summary Alcohol dependence and multiple withdrawal experiences are related to increased severity of alcohol use disorder (AUD), craving, and resistance to treatment. Alcohol abuse gains control over behavior, in part, through pathological adaptations of glutamatergic AMPA receptor (AMPAR) mechanisms that regulate synaptic and behavioral plasticity in brain reward pathways. The unique auxiliary protein, transmembrane AMPAR regulatory protein (TARP) γ-8, has been shown to regulate AMPAR trafficking, activity, and CaMKII-dependent plasticity, making it critical for AMPAR mediated neural transmission. An important feature of TARP γ-8 is its highly restricted expression limited to corticolimbic regions known to regulate glutamatergic response to alcohol including the medial prefrontal cortex (mPFC), basolateral amygdala (BLA), and the hippocampus, while noticeably absent from the nucleus accumbens (NAc). Evidence indicates chronic alcohol increases glutamate levels, which in turn promotes the influx of calcium, and initiating a cascade where CaMKII phosphorylates AMPARs to increase and sustain AMPAR activity. Since AMPAR activity is required for the development of new behavior (e.g., learning) and retention of actions (e.g., memory), this fundamental neural process may underlie the development, maintenance, and critically, dependence-escalated self-administration of alcohol. Therefore, this K99/R00 proposal will determine if TARP γ-8 regulates AMPAR mediated transmission in key brain regions during dependence-escalated alcohol self-administration. Aim 1 (K99 phase) of the proposal will investigate the role of TARP γ-8 dependent excitatory Ca2+ signaling in reward-related brain regions during alcohol self- administration in the mPFC, BLA, NAc, and vHPC using a highly novel multi-spectral, four-channel fiber photometry platform. Aim 2 (K99 phase) will examine TARP γ-8 as a mechanism of CIE vapor dependence- induced escalation of alcohol self-administration and the consequential co-localization of TARP γ-8 and AMPAR using confocal microscopy. Aim 3 (R00 phase) combines these techniques to evaluate Ca2+ signaling in key- reward brain regions during dependence-escalated alcohol self-administration. These findings are then extended by taking a circuit-based approach using a selective pharmacological manipulation in combination with fiber- photometry to evaluate site-specific TARP γ-8 bound AMPAR inhibition on “bottom-up” (BLA to NAc) and “top- down” (mPFC to NAc) Ca2+ signaling. This work moves the field forward by providing fundamental mechanistic insights into TARP γ-8 dependence-escalated alcohol self-administration which has high translational value for understanding and treating AUD and has the potential to inform development of new pharmacotherapeutic strategies that target AMPAR function in a highly-selective brain region specific manner.

项目摘要 酒精依赖和多次戒断经验与饮酒的严重程度增加有关 疾病（AUD），渴望和对治疗的抵抗力。酒精滥用会通过部分控制行为，部分地通过 调节突触和 大脑奖励途径中的行为可塑性。独特的辅助蛋白，跨膜AMPAR调节 蛋白质（TARP）γ-8已显示可调节AMPAR运输，活性和CAMKII依赖性可塑性， 使其对于AMPAR介导的神经传播至关重要。 TARPγ-8的一个重要特征是它的高度 限制的表达限制为已知调节谷氨酸剂反应的皮质脂质区域 包括培养基前额叶皮层（MPFC），副杏仁核（BLA）和海马，而 伏隔核（NAC）明显不存在。证据表明慢性酒精会增加谷氨酸 水平又促进了钙的影响，并启动了CAMKII磷酸化的级联反应 AMPAR提高和维持AMPAR活动。由于开发新的AMPAR活动是必需的 行为（例如，学习）和行动的保留（例如记忆），这种基本神经过程可能是基础的 依赖性提出的酒精的自我管理。所以， 该K99/R00建议将确定TARPγ-8是否调节关键大脑区域的AMPAR介导的传播 在依赖性提出的酒精自我管理期间。该提案的目标1（K99阶段）将调查 饮酒期间与奖励相关的大脑区域中TARPγ-8依赖性兴奋性Ca2+信号的作用 使用高度新颖的多光谱，四通道纤维在MPFC，BLA，NAC和VHPC中给药 光度法平台。 AIM 2（K99期）将检查TARPγ-8作为CIE蒸气依赖性的机制 - 诱导酒精自我给药的升级以及TARPγ-8和AMPAR的相应共定位 使用共聚焦显微镜。 AIM 3（R00相）结合了这些技术，以评估钥匙中的Ca2+信号传导 奖励依赖性提出的酒精自我管理期间的大脑区域。这些发现然后扩展 通过采用基于电路的方法，使用选择性药物操纵与纤维结合使用 评估位点特异性TARPγ-8 AMPAR抑制对“自下而上”（BLA至NAC）和“ Top-top- 下降”（MPFC至NAC）CA2+信号传导。这项工作通过提供基本的机制来向前移动该领域 对TARPγ-8依赖性提出的酒精自我给药的见解，该饮酒自给 理解和治疗AUD，并有可能为开发新的药物治疗而告知 以高度选择性的大脑区域特定方式靶向AMPAR的策略。