Genetic dissection of lateral habenula neuronal circuitry in cocaine seeking

 描述（由申请人提供）：这个K01指导研究科学家奖将提供支持和指导，以建立候选人，Sunila Nair博士作为分子和行为神经科学的独立研究人员。这个应用程序的培训部分将建立在我在行为神经药理学方面的专业知识，研究滥用药物和非药物成瘾者复发所涉及的神经回路，并进行分子生物学方面的培训。 神经科学，这将使我能够建立自己的尖端研究计划。我的高级科学培训将集中在两个关键领域：1）病毒介导的基因转移工具的构建和包装，以及2）分子遗传学培训。职业发展活动将由John Neumaier博士（主要导师）和Eric Turner博士（共同导师）指导，他们是拥有非重叠但相关专业知识和指导初级教师的特殊记录的研究人员。拟议的项目将在华盛顿大学精神病学和行为神经科学系进行，该系为高影响力的神经科学研究提供了一个特殊的环境。这是一个智力刺激和高度协作的环境，大多数高级教师都有指导初级调查人员的记录。研究部分将调查外侧缰核，这是一个研究不足的上丘脑核，可能在吸毒和寻求行为中起着关键作用。作为先前资助的NIH R21应用程序的一部分，我正在绘制控制外侧缰神经元活动并介导这些行为的神经回路。然而，解剖外侧缰及其靶点的精确功能性神经回路的一个主要障碍是，对这些神经元的遗传特性知之甚少。这个大脑区域在解剖学和功能上是异质的，要彻底了解外侧缰在成瘾（和其他疾病）中的作用， 对神经肽、神经递质、受体、摄取转运蛋白和其他在该脑区表达的靶点的完整理解。此外，事实上没有关于大鼠外侧缰亚区基因表达的特定模式的信息，以及在复杂的行为任务中是否发生了改变。我将使用最近开发的“Ribotag”技术结合DREADD的询问在可卡因服用和寻求过程中外侧缰及其相关神经元回路的mRNA翻译的变化。使用这种尖端的方法将使我能够检测在复杂的行为任务中，在外侧缰神经元中进行主动翻译的mRNA的变化，这些神经元投射到其关键目标。该K 01应用的科学目标首先是进行先进分子生物学和遗传学培训，并开发新工具来回答上述研究问题。其次，成功地实施这些工具，剖析遗传差异的作用，外侧缰在可卡因服用与寻求行为。第三，在大鼠模型中检查外侧缰中的通路特异性遗传差异，该模型包含药物使用的负面后果，其在临床上非常接近人类成瘾。

 DESCRIPTION (provided by applicant): This K01 Mentored Research Scientist Award will provide support and mentorship to establish the Candidate, Dr. Sunila Nair as an independent researcher in molecular and behavioral neuroscience. The training component of this application will build on my expertise in behavioral neuropharmacology investigating the neural circuits involved in relapse to drugs of abuse and non-drug reinforcers, with training in molecular neuroscience which will enable me to establish my own cutting-edge research program. My advanced scientific training will be focused in two key areas: 1) construction and packaging of viral-mediated gene transfer tools, and 2) training in molecular genetics. The career development activities will be mentored by Dr. John Neumaier (Primary Mentor) and Dr. Eric Turner (Co-Mentor), researchers that possess non-overlapping, but related expertise and exceptional records of mentoring junior faculty. The proposed project will take place in the Department of Psychiatry and Behavioral Neuroscience at the University of Washington, which provides an exceptional environment for high-impact neuroscience research. It is an intellectually stimulating and highly collaborative environment and most senior faculty members have a track record for mentoring junior investigators. The research component will investigate the lateral habenula, an understudied epithalamic nucleus that likely plays a key role in drug taking and seeking behaviors. As part of a previously funded NIH R21 application, I am mapping the neurocircuitry that controls lateral habenula neuronal activity and mediates these behaviors. However, a major roadblock in dissecting the precise functional neurocircuitry of the lateral habenula and its targets is the fact that very little is known about the genetic properties of thes neurons. This brain region is anatomically and functionally heterogeneous and a thorough understanding of the role of the lateral habenula in addiction (and other disorders) will require a complete understanding of the neuropeptides, neurotransmitters, receptors, uptake transporters and other targets that are expressed in this brain region. Further, there is virtually no informatin on the specific patterns of gene expression in lateral habenula sub-regions in rats and if this is altered during complex behavioral tasks. I will use the recently developed `Ribotag' technology in conjunction with DREADD's to interrogate changes in mRNA translation in the lateral habenula and its associated neuronal circuitry during cocaine taking and seeking. The use of this cutting-edge approach will allow me to detect changes in mRNA undergoing active translation in lateral habenular neurons projecting to its key targets during complex behavioral tasks. The scientific goals for this K01 application are firstly, to train in advanced molecular biology and genetics and develop novel tools to answer the above research questions. Secondly, to successfully implement these tools to dissect genetic differences in the role of the lateral habenula in cocaine-taking versus seeking behaviors. Thirdly, to examine pathway-specific genetic differences in lateral habenula in a rat model that incorporates the negative consequences of drug use that closely approximates addiction clinically in humans.