Contributions of Glial Glutamate Transport and NMDA Receptors in Nicotine Relapse

胶质细胞谷氨酸转运和 NMDA 受体在尼古丁复吸中的作用

基本信息

批准号：
9301729
负责人：
Cassandra D Gipson-Reichardt
金额：
$ 2.1万
依托单位：
ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-01-15 至 2018-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9301729
关键词：
Adult Affect Alkaloids Animals Award Biological Assay Brain Brain region Caliber Ceftriaxone Cells Cessation of life Chemosensitization Cues Data Dendritic Spines Dependency Development Down-Regulation Drug Addiction Electrophysiology (science)Exposure to Glutamate Transporter Glutamates Head Health Intravenous Learning Measures Mediating Mentors Methods Microinjections Morphology N-Methyl-D-Aspartate Receptors N-Methylaspartate National Research Service Awards Neurobiology Neuroglia Neurons Nicotine Nicotine Dependence Nucleus Accumbens Outcome Pharmaceutical Preparations Pharmacotherapy Phase Procedures Property Proteins Rattus Relapse Research Risk Factors Role Saline Self Administration Self-Administered Slice Small Interfering RNA Smoke Smoking Smoking Behavior Synapses Synaptic plasticity Techniques Tissues Tobacco Tobacco Dependence Tobacco smoking Training Treatment Efficacy Up-Regulation Vertebral column Western Blotting cigarette smoking design drug abstinence drug development drug reward innovation knock-down neurobiological mechanism neurochemistry neuropsychiatric disorder nicotine abuse novel novel therapeutics patch clamp prevent programs protein function receptor reduce tobacco use research study skills treatment group uptake

项目摘要

DESCRIPTION (provided by applicant): Nicotine abuse and addiction represents a large health liability. Indeed, cigarette smoking-related illness results in an estimated 6,000,000 deaths per year worldwide, yet 20% of adults currently smoke, and among those who attempt to quit, >90% relapse. Nicotine, the primary active alkaloid in tobacco, is self-administered by animals and produces cellular adaptations in brain regions associated with drug reward, such as the nucleus accumbens. Due to the cue dependency of smoking behavior, exposure to nicotine-associated cues is a risk factor for relapse. Here, I examine the role of glial glutamate transport and NMDA receptors in nicotine relapse vulnerability. I have found that nucleus accumbens core glutamatergic mechanisms are involved in nicotine relapse, including increased synaptic strength (measured as increased spine diameter and AMPA currents) and accompanying protein changes (including a decrease in the glial glutamate transporter, GLT1, and increases in the AMPA subunit GluA1 and NMDA subunit GluN2B). During the proposed award period, I will explore the mechanisms mediating cued nicotine reinstatement and a possible neuron-glia interaction underlying relapse vulnerability. In the K99 aims, I propose to functionally characterize nicotine-mediated down-regulation of GLT1, and to determine its role in reinstated nicotine seeking. To do this, I will learn glutamate uptake and whole cell patch clamp electrophysiology strategies, as well as employ my Western blot and intracranial microinjection skills I acquired during my F32 NRSA. I will also use antisense vivo morpholinos and ceftriaxone to examine the impact of up- or down-regulated GLT1 on reinstatement of nicotine seeking. I will employ these techniques during the R00 period to further characterize the role of GluN2B in cue-reinstated nicotine seeking, and to explore a potential neuron-glia interaction mediating nicotine relapse. During the R00 period, I will electro physiologically determine if the unregulated GluN2B receptors I found in nicotine-extinguished animals are extra synaptic and necessary for cued nicotine seeking. I will accomplish this with an innovative set of techniques including a coagonist degradation procedure using whole cell patch clamp, and administration of siRNA constructs to down regulate GluN2B in nicotine-extinguished animals to determine if normalizing this protein inhibits cued nicotine seeking in nicotine-extinguished animals. This will indicate a key role of glutamate overflow and activation of extra synaptic NMDA receptors in relapse vulnerability. Next, I will determine if restoring GluN2B with siRNA or GLT1 with ceftriaxone indirectly restores GLT1 or GluN2B, respectively, indicating a neuron-glia interaction in cue-induced nicotine relapse. Finally, I will examine if restoring GluN2B or GLT1 prevents the rapid, transient synaptic plasticity I previously found during cued nicotine reinstatement. These experiments have the potential to reveal novel neurobiological mechanisms of nicotine addiction, and could contribute to the development of novel therapeutic options aimed at reversing nicotine-induced neurobiological alterations.

描述（由申请人提供）：尼古丁滥用和成瘾代表了巨大的健康责任。实际上，与吸烟有关的疾病每年估计每年有6,000,000人死亡，但目前有20％的成年人吸烟，而在试图戒烟的患者中，复发> 90％。尼古丁是烟草中的主要活性生物碱，是由动物自我管理的，并在与药物奖励相关的大脑区域（例如伏隔核）中产生细胞适应性。由于吸烟行为的提示依赖性，接触尼古丁相关的提示是复发的危险因素。在这里，我检查了神经胶质谷氨酸转运的作用尼古丁复发脆弱性中的NMDA受体。 I have found that nucleus accumbens core glutamatergic mechanisms are involved in nicotine relapse, including increased synaptic strength (measured as increased spine diameter and AMPA currents) and accompanying protein changes (including a decrease in the glial glutamate transporter, GLT1, and increases in the AMPA subunit GluA1 and NMDA subunit GluN2B).在拟议的奖励期间，我将探索介导的尼古丁恢复原状的机制，以及可能复发脆弱性的神经元 - 胶体相互作用。在K99的目的中，我建议在功能上表征尼古丁介导的GLT1下调，并确定其在恢复尼古丁寻求的作用。为此，我将学习谷氨酸摄取和全细胞斑块夹电生理学策略，并采用我在F32 NRSA中获得的蛋白质印迹和颅内微分注射技能。我还将使用反义的体内形态和头孢曲松来检查上调或下调的GLT1对恢复尼古丁寻求的影响。我将在R00期间采用这些技术来进一步表征Glun2b在提示提示的尼古丁寻求中的作用，并探索介导尼古丁复发的潜在神经元 - 胶体相互作用。在R00期间，我将在生理学上确定我在尼古丁效果的动物中发现的未受监管的Glun2b受体是否是额外的突触，并且对于提示尼古丁寻求的必要。我将通过一套创新的技术来完成此操作，包括使用全细胞贴片夹的辅助降解程序，以及对烟碱效果的siRNA构建体的施用，以降低尼古丁效果的动物中的Glun2b，以确定该蛋白抑制这种蛋白质是否抑制了氨基氨基氨基氨基氨基氨基氨基氨基氨基氨基氨基调的动物。这会表明谷氨酸溢流并激活额外的突触NMDA受体在复发脆弱性中的关键作用。接下来，我将确定用与头孢曲松中的siRNA或GLT1恢复间接恢复GLT1或GLUN2B的GLUN2B，表明在提示诱导的尼古丁复发中神经元 - 葡萄球菌相互作用。最后，我将检查恢复GLUN2B或GLT1是否可以防止我在提示尼古丁恢复原状过程中先前发现的快速，短暂的突触可塑性。这些实验具有揭示尼古丁成瘾的新型神经生物学机制的潜力，并可能有助于发展新型治疗方案，旨在逆转尼古丁引起的神经生物学改变。