Molecular Regulation of GABAalpha Function in Amygdala

杏仁核 GABAα 功能的分子调控

• 批准号: 7276612
• 负责人: Kerry J Ressler
• 金额: $ 36.92万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2011-08-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7276612
• 关键词: Acute; Adult; Affect; Alcohol or Other Drugs use; Alcohols; Alleles; Amygdaloid structure; Anti-Anxiety Agents; Anticonvulsants; Anxiety; Autoradiography; Behavioral; Benzodiazepines; Binding; Biological; Brain; Cell Nucleus; Chemistry; Chromosome Deletion; Chromosome Pairing; Chronic; Cocaine; Comorbidity; Complex; Cues; Data; Dependence; Diazepam; Disease; Down-Regulation; Electrophysiology (science); Event; Flunitrazepam; Fright; Functional disorder; Gene Proteins; Gene Silencing; Genes; Immunoblotting; In Situ Hybridization; In Vitro; Injection of therapeutic agent; Lead; Measures; Mediating; Molecular; Molecular Genetics; Morphine; Mus; Nicotine; Nucleus Accumbens; Pharmaceutical Preparations; Phase; Property; Regulation; Relapse; Research Personnel; Role; Secondary to; Self Administration; Small Interfering RNA; Stress; Structure; Subfamily lentivirinae; Substance Addiction; Substance abuse problem; Substance of Abuse; Synapses; System; Testing; Withdrawal; addiction; behavior measurement; biological adaptation to stress; conditioned fear; drug craving; drug of abuse; dual diagnosis; gamma-Aminobutyric Acid; gephyrin; insight; lentiviral-mediated; neurotensin mimic 2; novel strategies; programs; receptor; receptor expression; receptor function; recombinase; response; sedative; transmission process; zolpidem

DESCRIPTION (provided by applicant): Comorbid anxiety may predispose addicts to relapse secondary to the biological effects of stress and anxiety on the brain. Furthermore 'hyperexcitability' of the amygdala, as repeatedly seen during stress or fear and substance withdrawal, may contribute to anxiety responses and drug craving. Such increased excitability within the basolateral amygdala (BLA) may be due in part to decreased GABAA activation following stress or substance use. These studies will examine the molecular mechanisms of activity- dependent regulation of GABAA receptors by focusing on fear conditioning and benzodiazepine (BZD) dependence. Both of these manipulations lead to downregulation of GABAA receptors in the BLA. We will compare the molecular mechanisms of GABAA regulation following the stress of fear conditioning with acute, chronic, and withdrawal phases of BZD administration. We hypothesize that: 1) activity-dependent modulation of GABAA within the basolateral amygdala is a common mechanism of both substance dependence and stress response; and 2) GABAergic manipulations that affect BZD response will also affect acquisition and expression of conditioned fear. In this proposal, we will compare the molecular, behavioral, and electrophysiological mechanisms of GABAA regulation following conditioned fear stress with acute, chronic, and withdrawal phases of benzodiazepine administration. We propose that these pharmacological and behavioral events share similar mechanisms involving activity-dependent downregulation of the GABAA complex. A variety of molecular genetic approaches in mice will be used to test this hypothesis including: 1) examination of GABAA complex genes and proteins with in situ hybridization and immunoblotting; 2) manipulation of GABAA receptor expression via amygdala-specific deletion of the GABAA alpha1 gene using the Cre/lox system; 3) manipulation of post-synaptic GABAA alpha2 gene using Lentiviral-mediated gene silencing; 4) manipulation of receptor clustering through silencing of the Gephyrin gene. These studies have direct implications for mechanisms of benzodiazepine dependence, withdrawal, and relapse. Given the role of amygdala GABAergic systems in modulating dopaminergic tone within the nucleus accumbens, amygdala GABAA regulation may have a more general role in addictive disorders. In fact, recent studies have suggested that morphine, alcohol, cocaine and nicotine may also alter GABAA functioning in amygdala, and acutely enhancing GABA levels blocks reinstatement by drug-associated cues or self-administration. This proposal represents a novel approach to understanding shared molecular and cellular mechanisms that may mediate comorbid anxiety and substance abuse disorders. Understanding the mechanisms shared by stress and drug response is critical for understanding pathophysiology and proposing new treatments for these highly comorbid disorders.

描述(由申请者提供)：由于压力和焦虑对大脑的生物影响，共病焦虑可能使吸毒者容易复发。此外，杏仁核的“过度兴奋”，就像在压力或恐惧和药物戒断时反复出现的那样，可能会导致焦虑反应和药物渴望。杏仁基底外侧核(BLA)兴奋性的增加可能部分是由于应激或药物使用后GABAA激活减少所致。这些研究将通过恐惧条件反射和苯二氮(BZD)依赖来研究GABAA受体活性依赖调节的分子机制。这两种操作都导致BLA中GABAA受体的下调。我们将比较恐惧条件反射应激后GABAA调节的分子机制与BZD给药的急性、慢性和戒断阶段。我们假设：1)杏仁基底外侧核内GABAA的活动依赖性调节是物质依赖和应激反应的共同机制；2)影响BZD反应的GABA能操作也会影响条件性恐惧的获得和表达。在这项建议中，我们将比较条件恐惧应激后GABAA调节的分子、行为和电生理机制与苯二氮卓类药物的急性、慢性和停药阶段。我们认为，这些药理和行为事件具有类似的机制，涉及GABAA复合体的活性依赖下调。将使用多种分子遗传学方法在小鼠中验证这一假说，包括：1)用原位杂交和免疫印迹法检测GABAA复合体基因和蛋白质；2)通过CRE/LOX系统通过杏仁核特异缺失GABAAα1基因来操纵GABAA受体的表达；3)通过慢病毒介导的基因沉默来操纵突触后的GABAAα2基因；4)通过沉默geporrin基因来操纵受体聚集。这些研究对苯二氮类药物依赖、戒断和复发的机制有直接的影响。鉴于杏仁核GABA能系统在调节伏隔核多巴胺能张力中的作用，杏仁核GAAA调节在成瘾障碍中可能具有更普遍的作用。事实上，最近的研究表明，吗啡、酒精、可卡因和尼古丁也可能改变杏仁核中GABAA的功能，而急剧提高的GABA水平会阻止药物相关线索或自我给药的恢复。这一建议代表了一种新的方法来理解共同的分子和细胞机制，这些机制可能调节共病的焦虑和物质滥用障碍。了解应激和药物反应的共同机制对于了解病理生理学和为这些高度共存的疾病提出新的治疗方法至关重要。