Genetic dissection of lateral habenula neuronal circuitry in cocaine seeking

 描述(由申请者提供)：K01指导研究科学家奖将提供支持和指导，以确立候选人Sunila Nair博士为分子和行为神经科学的独立研究员。这个应用程序的培训部分将建立在我的行为神经药理学专业知识的基础上，研究滥用药物和非药物增强剂复发所涉及的神经回路，并进行分子培训 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，2)分子遗传学方面的培训。职业发展活动将由John Neumaier博士(主要导师)和Eric Turner博士(联合导师)指导，这两位研究人员拥有不重叠但相关的专业知识和指导初级教师的杰出记录。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。这是一个智力激发和高度合作的环境，大多数资深教员在指导初级调查人员方面都有过往记录。研究部分将调查外侧缰核，这是一个未被充分研究的上皮核，可能在吸毒和寻找行为中发挥关键作用。作为之前资助的NIH R21应用程序的一部分，我正在绘制控制外侧缰核神经元活动并调节这些行为的神经回路。然而，解剖外侧缰核及其靶点的精确功能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。这一大脑区域在解剖和功能上是不同的，彻底了解外侧缰核在成瘾(和其他障碍)中的作用将需要 完全了解在这个大脑区域表达的神经肽、神经递质、受体、摄取转运体和其他靶标。此外，几乎没有关于大鼠外侧缰核亚区基因表达的特定模式的信息，以及这种模式是否在复杂的行为任务中发生改变。我将使用最近开发的‘Ribotag’技术结合DREADD‘s来询问在吸食和寻找可卡因过程中外侧缰核及其相关神经元电路中信使核糖核酸翻译的变化。这种尖端方法的使用将使我能够检测在复杂的行为任务中投射到关键目标的外侧缰核神经元中正在进行主动翻译的mRNA的变化。K01应用程序的科学目标是：首先，培训高级分子生物学和遗传学，并开发新的工具来回答上述研究问题。其次，为了成功地使用这些工具来剖析外侧缰核在吸食可卡因和寻找可卡因行为中所起的作用的遗传差异。第三，在一个大鼠模型中检查外侧缰核中特定路径的遗传差异，该模型结合了药物使用的负面后果，在临床上与人类的成瘾非常接近。

 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.