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

基本信息

批准号：
8127798
负责人：
Caroline Evelyn Bass
金额：
$ 14.61万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
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/8127798
关键词：
Ablation Addictive Behavior Address Adult Area Bass Behavior Behavioral Behavioral Model Biology Brain Brain region CNR1 gene Cannabinoids Catalepsy Cocaine Complex Corpus striatum structure Development Dopamine Dopamine D1 Receptor Dopamine D2 Receptor Dopamine Receptor Dorsal Drug Addiction 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 meetings natural hypothermia neurotransmission neurotransmitter release novel programs protein expression receptor receptor function research study skills small hairpin RNA tool vector

项目摘要

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）：该 K01 指导研究职业发展奖旨在培训维克森林大学生理学与药理学系初级教师 Caroline E. Bass，开发一个独立的假设驱动研究项目来调查成瘾过程由中皮质边缘系统中的多巴胺和大麻素受体调节。 Bass 博士将运用她在开发用于传递 RNA 干扰 (RNAi) 转基因的腺相关病毒 (AAV) 载体方面的技能来开发自己的项目，重点是在战略性定义的大脑位点中敲低 D1、D2 和 CB1 受体，这对可卡因增强功效的影响，以及卡纳宾和多巴胺系统如何在可卡因自我给药模型中相互作用。 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 博士，她在行为模型（特别是大鼠自我给药）方面的指导将来自 David C.S. Roberts 博士，他是这些领域的杰出科学家。各自的领域。其他指导将涵盖与学术晋升和国家认可相称的专业技能的各个方面。为了实现本提案的目标，提出了以下具体目标： 1) 生成并表征含有 RNAi 表达盒的 AAV 载体，这些表达盒会引起大鼠 D1、D2 和 CB1 受体 mRNA 的降解，并确定多巴胺和 CB1 分子成分的补偿性变化。这种操作产生的内源性大麻素神经传递； 2) 通过将 CB1 RNAi-AAV 注射到纹状体和髓质中，研究 THC 非条件行为效应的作用位点，并评估 THC 在诱导镇痛、活动减退、体温过低和僵住方面的功效； 3) 通过在大鼠自我给药可卡因中进行位点特异性注射 D1 或 D2 RNAi-AAV，描绘和定义腹侧纹状体、背侧纹状体和 VTA 中的 D1 和 D2 受体在可卡因增强作用中的作用； 4) 表征腹侧纹状体、背侧纹状体和 VTA 的 CB1 受体消融对可卡因自我给药的影响。具体来说，CB1 RNAi-AAV 将被注射到这些区域，并评估可卡因的增强功效。在这些目标的所有阶段，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量中相互作用。这些研究将解决我们对多巴胺受体强化功能知识的根本差距，并作为了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出新颖且更有效的药物成瘾治疗干预措施。