M4 muscarinic acetylcholine receptor signaling as a potent regulator of motor deficits

候选人：我的长期目标是成为一名独立的调查员， 合作研究团队，以了解神经系统疾病的机制和新的治疗方法。 具体来说，我的兴趣围绕着了解不同来源的乙酰胆碱（ACh，即胆碱能 interneurons vers pedunculopontine nucleus）调节运动功能障碍，以及如何不平衡 多巴胺（DA）和M4毒蕈碱乙酰胆碱受体（mAChR）信号传导加剧运动功能障碍。我 在生物化学和电生理学方面有很强的背景。我打算学习行为和伏安法 技术，以完善我的培训，并有必要的技能，以产生高影响力的出版物， 成功提交R 01。我拿到了博士学位。2015年4月，这是我获得K99/R 00资格的最后一年。 培训：除了康恩博士，我还有一个行为药理学专家咨询委员会。 琼斯，肌张力障碍和多巴胺能信号，埃利希博士，并在使用分子探针了解GPCR 信号林斯利博士该委员会将提供必要的培训和指导，以实现这一目标 提议在委员会之外，我们已经确定，无论是在范德比尔特和外部，课程，研讨会， 和会议，以提供进一步的技术培训，演讲经验，负责任的行为研究， 和必要的技能（提供谈判，任期，实验室管理等）过渡到独立。 研究：抗mAChR治疗在减少某些运动障碍的运动症状方面有效， 严重的副作用限制了它们的效用。我们的实验室在阐明mAChR的作用方面取得了突破性进展， 发现M4 mAChR亚型减少DA释放，信号传导和相关运动行为。此外，M4 DA信号传导的抑制在纹状体外紧张性地发生。这让我们想到了一个模型， 当M4释放或信号传导较低时，这使得M4信号传导占主导地位，导致运动功能障碍，并导致 我们的广泛假设是M4拮抗剂将减少运动障碍中的运动缺陷。当M4 活性可能是DA和基底神经节活性的关键调节剂，提供与非选择性 没有副作用的mAChRs，这还没有经过测试。在初步数据中，我们报告了新工具的发现 化合物直接测试我们的假设，证明失去D1 DA受体信号转导减少基础 神经节活动并产生运动缺陷，M4活动双向调节运动缺陷。在目标1中， 使用预测抗帕金森病疗效的动物模型，我们将测试乙酰胆碱如何，以及通过什么来源， M4活动调节运动缺陷。在目标2中，我们将使用与以下相关的D1 DA受体信号转导的损失模型： 肌张力障碍，以测试减少的DA信号，可能允许M4信号占主导地位，如何影响基础 神经节输出和信号传导。在目标3中，我们将测试基底神经节肌张力障碍如何以及在何处与D1丢失相关。 DA受体信号传导导致运动缺陷，以及M4如何调节这些行为。

Candidate: My long term goal is to become an independent investigator running an interdisciplinary and collaborative research team to understand the mechanisms and novel treatments of neurological disorders. Specifically, my interests surround understanding how different sources of acetylcholine (ACh, i.e. cholinergic interneurons verse pedunculopontine nucleus) regulate motor dysfunction, as well as how imbalanced dopamine (DA) and M4 muscarinic acetylcholine receptor (mAChR) signaling exacerbates motor dysfunction. I have a strong background in biochemistry and electrophysiology. I propose to learn behavioral and voltammetry techniques to round out my training, and have the necessary skills to produce high impact publications and successful R01 submissions. I received my Ph.D. in April 2015, this is my last year of eligibility for the K99/R00. Training: In addition to Dr. Conn, I have an advisory committee of experts in behavioral pharmacology, Dr. Jones, dystonia and dopaminergic signaling, Dr. Ehrlich, and in using molecular probes to understand GPCR signaling, Dr. Lindsley. This committee will provide the necessary training and guidance to accomplish this proposal. Outside of the committee, we have identified, both at Vanderbilt and externally, courses, seminars, and meetings to provide further technical training, presentation experience, responsible conduct in research, and the necessary skills (offer negotiations, tenure, lab management, etc.) to transition to independence. Research: Anti-mAChR therapy is efficacious at reducing motor symptoms of some movement disorders, but severe side effects limit their utility. Our lab has made breakthroughs elucidating the roles of mAChRs, and found the M4 mAChR subtype diminishes DA release, signaling, and related motor behaviors. Additionally, M4 inhibition of DA signaling occurs tonically outside the striatum. This has led us to the model that when DA release or signaling is low, this allows M4 signaling to predominate, leading to motor dysfunction, and has led to our broad hypothesis that M4 antagonists will reduce motor deficits in movement disorders. While M4 activity may be a critical modulator of DA and basal ganglia activity, providing similar efficacy to non-selective mAChRs without side effects, this has not been tested. In preliminary data, we report the discovery of novel tool compounds to directly test our hypothesis, demonstrate losing D1 DA receptor signaling diminishes basal ganglia activity and produces motor deficits, and M4 activity bi-directionally modulates motor deficits. In aim 1, using animal models predictive of anti-parkinsonian efficacy, we will test how, and through what ACh sources, M4 activity modulates motor deficits. In aim 2, we will use a model of loss of D1 DA receptor signaling linked to dystonia to test how diminished DA signaling, possibly allowing M4 signaling to predominate, effects basal ganglia output and signaling. In aim 3, we will test how and where in the basal ganglia dystonia linked loss of D1 DA receptor signaling leads to motor deficits, and how M4 modulates these behaviors.