CB1 and DA Receptor Deletion by RNAi in Cocaine Self-Administration

可卡因自我给药中通过 RNAi 删除 CB1 和 DA 受体

• 批准号: 7588997
• 负责人: Caroline Evelyn Bass
• 金额: $ 13.81万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-30 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7588997
• 关键词: Ablation; Addictive Behavior; Address; Adult; Area; Bass; Behavior; Behavioral; Behavioral Model; Biology; Brain; Brain region; CNR1 gene; Cannabinoids; Catalepsy; Cocaine; Complex; Condition; Corpus striatum structure; Development; Dopamine; Dopamine D2 Receptor; Dopamine Receptor; Dorsal; Drug Addiction; Drug Regulations; Drug abuse; Endocannabinoids; Faculty; Financial compensation; Foundations; Gene Expression; Goals; Individual; Injection of therapeutic agent; Intake; Investigation; K-Series Research Career Programs; Knockout Mice; Knowledge; Lead; Learning; Memory; Mentors; Messenger RNA; Modeling; Molecular; Monoacylglycerol Lipases; Mus; Neurons; Outcome; Pharmacology; Physiology; Population; Principal Investigator; Process; Protein Analysis; Proteins; Psychological reinforcement; RNA Interference; RNA Viruses; Rattus; Recombinant adeno-associated virus (rAAV); Regulation; Research; Research Project Grants; Resolution; Rodent; Role; Schedule; Scientist; Self Administration; Self-Administered; Site; Social Interaction; Staging; System; Techniques; Therapeutic Intervention; Time; Tissues; Training; Transcript; Transgenes; Tyrosine 3-Monooxygenase; Ventral Striatum; Ventral Tegmental Area; Viral Vector; Virus; addiction; base; cannabinoid receptor; defined contribution; design; dopamine system; dopamine transporter; drug reinforcement; drug seeking behavior; fatty acid amide hydrolase; feeding; forest; improved; in vivo; innovation; interest; knock-down; natural hypothermia; neurotransmission; neurotransmitter release; novel; programs; protein expression; receptor; receptor function; research study; skills; small hairpin RNA; tool; vector

DESCRIPTION (provided by applicant): This K01 Mentored Research Career Development Award is to train Caroline E. Bass, junior faculty in the Dept. Physiology & Pharmacology at Wake Forest Univ., to develop an independent hypothesis-driven research project to investigate addiction processes that are modulated by dopamine and cannabinoid receptors in the mesocorticolimbic system. Dr. Bass will apply her skills in the development of adenoassociated virus (AAV) vectors for the delivery of RNA interference (RNAi) transgenes to develop her own project focused on the knock down of D1, D2 and CB1 receptors in strategically defined brain loci, the impact this has on the reinforcing efficacy of cocaine, and how the cananbiniod and dopamine systems interact in a cocaine self-administration model of drug abuse. Dr. Bass's mentoring in cannabinoid pharmacology and gene expression changes will come from Dr. Allyn C. Howlett, and her mentoring in the behavioral models, particularly rat self-administration, will come from Dr. David C.S. Roberts, who are distinguished scientists in these respective areas. Other mentoring will cover all aspects of professional skills commensurate with academic promotion and national recognition. The following specific aims are proposed to meet the goals of this proposal: 1) Generate and characterize AAV vectors containing RNAi expression cassettes that cause the degradation of D1, D2 and CB1 receptor mRNA in rats and determine the compensatory changes in molecular components of dopamine and endocannabinoid neurotransmission resulting from this manipulation; 2) Investigate the site(s) of action of non-conditioned behavioral effects of THC by injecting the CB1 RNAi-AAV into the striatum and medulla and assessing the efficacy of THC in inducing antinociception, hypoactivity, hypothermia, and catalepsy; 3) delineate and define the contribution of D1 and D2 receptors in the ventral striatum, dorsal striatum and VTA in the reinforcing effects of cocaine by site specific injection of D1 or D2 RNAi-AAV in rats self-administering cocaine; 4) Characterize the impact of CB1 receptor ablation from the ventral striatum, dorsal striatum and VTA on cocaine self-administration. Specifically, the CB1 RNAi-AAV will be injected into these regions and the reinforcing efficacy of cocaine will be assessed. During all stages of these aims the compensatory mechanisms in cannabinoid and dopamine neurotransmission will be assessed to determine how these receptor systems are interacting in modulating cocaine intake. These studies will address fundamental gaps in our knowledge of dopamine receptor function in reinforcement and serve as a basis for understanding how cannabinoid and dopamine receptor systems interact to regulate addictive behaviors. These findings may lead to the development of novel and more effective therapeutic interventions for drug addiction.

描述（由申请人提供）：这个K 01指导研究职业发展奖是培养卡罗琳E。巴斯，系里的初级教员。维克森林大学的生理学和药理学，开发一个独立的假设驱动的研究项目，以调查成瘾过程中，调节多巴胺和大麻素受体在中皮层边缘系统。巴斯博士将运用她在腺相关病毒（AAV）载体的开发中的技能，用于RNA干扰（RNAi）转基因的传递，以开发她自己的项目，重点是在战略性定义的大脑位点中敲除D1，D2和CB 1受体，这对可卡因的增强功效的影响，以及cananbiniod和多巴胺系统如何在可卡因自我管理药物滥用模型中相互作用。Bass博士在大麻素药理学和基因表达变化方面的指导将来自Allyn C. Howlett和她在行为模型方面的指导，特别是大鼠自我管理，将来自大卫博士。罗伯茨，谁是杰出的科学家在这些各自的领域。其他辅导将涵盖与学术晋升和国家承认相称的专业技能的所有方面。1）产生和表征含有RNAi表达盒的AAV载体，所述RNAi表达盒在大鼠中引起D1、D2和CB 1受体mRNA的降解，并确定由这种操作引起的多巴胺和内源性大麻素神经传递的分子组分的补偿性变化; 2）通过将CB 1 RNAi-AAV注射到纹状体和髓质中来研究THC的非条件行为效应的作用位点，并评估THC在诱导抗伤害感受、活动减退、体温过低和僵住症中的功效; 3）通过在可卡因自我给药的大鼠中定位注射D1或D2 RNAi-AAV，描绘并确定腹侧纹状体、背侧纹状体和VTA中的D1和D2受体在可卡因增强效应中的贡献; 4）表征来自腹侧纹状体、背侧纹状体和VTA的CB 1受体消融对可卡因自我施用的影响。具体地，将CB 1 RNAi-AAV注射到这些区域中，并评估可卡因的增强功效。在这些目标的所有阶段中，将评估大麻素和多巴胺神经传递的补偿机制，以确定这些受体系统如何在调节可卡因摄入中相互作用。这些研究将解决我们对多巴胺受体在强化中的功能的认识中的根本差距，并作为理解大麻素和多巴胺受体系统如何相互作用以调节成瘾行为的基础。这些发现可能会导致开发新的和更有效的药物成瘾治疗干预措施。