Acetylcholinergic Neurotransmission During Aging

衰老过程中的乙酰胆碱能神经传递

• 批准号: 8967731
• 负责人: Hakeem O Lawal
• 金额: $ 13.91万
• 依托单位: DELAWARE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8967731
• 关键词: Acetylcholine; Action Potentials; Address; Adult; Affect; Age; Aging; Alabama; Alleles; Alzheimer' s Disease; Animals; Behavior; Behavioral; Behavioral Mechanisms; Biochemical; Biological Models; Biological Process; Brain; Carrier Proteins; Cell Culture Techniques; Centers of Research Excellence; Cholinergic Agonists; Cognitive deficits; Complex; Coupled; Cultured Cells; Cytoplasm; Defect; Delaware; Development; Dopamine; Drosophila genus; Drosophila melanogaster; Electrophysiology (science); Elements; Endocytosis; Faculty; Fractionation; Functional disorder; Funding; Future; Genes; Genetic; Genetic Techniques; Goals; Human Resources; Impaired cognition; In Vitro; Learning; Longevity; Mammals; Mediating; Melissa; Memory; Mentored Research Scientist Development Award; Mentors; Mentorship; Modeling; Molecular; Mus; Mutation; Myasthenia Gravis; Nerve Degeneration; Nervous system structure; Neurodegenerative Disorders; Neurons; Neurosciences; Neurotransmitters; Nicotine; Nicotine Dependence; Olfactory Learning; Organism; Parkinson Disease; Performance; Pharmacology; Physiological; Physiology; Play; Point Mutation; Positioning Attribute; Postdoctoral Fellow; Predisposition; Process; Research; Resources; Role; Secure; Series; Signal Transduction; Site; Sorting - Cell Movement; Specialist; Staging; Synapses; Synaptic Vesicles; System; Techniques; Testing; Time; Toxic Environmental Substances; Training; United States National Institutes of Health; Universities; Ursidae Family; acetylcholine transporter; addiction; age related; aged; base; brain tissue; career; cholinergic; cholinergic neuron; cholinergic synapse; classical conditioning; cognitive function; dopaminergic neuron; fly; gene environment interaction; graduate student; human disease; improved; in vivo; loss of function mutation; middle age; mutant; nervous system disorder; neurogenetics; neurotransmission; normal aging; novel therapeutics; overexpression; pathological aging; presynaptic; professor; programs; public health relevance; research study; response; skills; small molecule; synaptic function; tool; trafficking; vesicular monoamine transporter

 DESCRIPTION (provided by applicant): My career objectives over the next five years are to obtain R01 funding, secure tenure at Delaware State University and develop a research niche in using Drosophila to elucidate the role of graded changes in acetycholine release on physiology and behavior during aging. As a graduate student with Janis O'Donnell (U. Alabama) and a post-doctoral fellow with David Krantz (UCLA), I received extensive training in Drosophila neurogenetics and on the study of neuronal processes relevant to human diseases. In addition, with Dr. Krantz, an expert in the study of vesicular transporters and their role in neurotransmission, I have developed my own expertise in the function of vesicular neurotransmitter transporters. My most recent projects have involved using Drosophila transporter to test gene-environment interactions that may contribute to the susceptibility to Parkinson's disease (PD). In particular, I determined that overexpression of the vesicular monoamine transporter (VMAT) can protect dopamine neurons from environmental toxins by sequestering cytosolic dopamine away from its site of action. To build on the potential neuroprotective role for VMATs, I conducted a screen for small molecules capable of increasing its function and I successfully identified a potentially new class of aminergic agents. I will continue to study the neurogenetics of Drosophila vesicular neurotransmitter transporters in the early stages of my career as an independent faculty, but change my primary focus from VMAT to the Drosophila vesicular acetylcholine transporter (VAChT). Under Dr. Krantz's excellent mentorship, I expanded my repertoire of skills to include cell culture, behavioral analyses, biochemical fractionation and pharmacology. I am now exploiting this background to determine the biological function for VAChT in my own lab as a PI. I have developed a rigorous training plan that will help me to achieve my career transition into the study of cholinergic neurotransmission during aging. One important aspect of my training during the K01 award will be to learn electrophysiology. I have received basic training from Diane O'Dowd, a specialist in adult Drosophila brain electrophysiology. I learned how to prepare fly brain tissue and identify Drosophila cholinergic neurons. In addition, I received input on general electrophysiological concepts and techniques from Felix Schweizer at UCLA. Under these two mentors, I was able to establish the basics of patching onto a neuron and recording action potentials. I now plan to master these foundational skills and become an expert. Both Drs. Schweizer and O'Dowd have offered to continue to support me with their expertise. Importantly, Dr. Melissa Harrington at Delaware State has offered to enhance my training and will serve as my hands-on primary physiology mentor. She has given me full access to her rig and under her mentorship I will continue to hone my skills. She has also provided substantial input into the current application. I recently started my own lab as an Assistant Professor in the NIH/COBRE Neuroscience program at Delaware State. I received a start-up package of $320,000. This support has afforded me valuable resources/personnel to achieve my goals (see Training Plan). My goal is to become an expert in using electrophysiology to investigate how differences in ACh release effect synaptic function. I will couple this skill set with my current expertise to determine how te changes in the function of cholinergic synapses may correlate with behavioral decline during aging. Moreover, I will be well-positioned to address fundamental questions about ACh release, behavior and aging that have clear translational significance. To study these issues, I will use an allelic series of point mutants in the Drosophila VAChT, the protein that packages and transports ACh for synaptic release. These alleles which range from mild to severe, and will allow me to determine the relationship between graded changes in acetylcholine release and progressive deficits in acetylcholine-mediated behaviors such as learning. The short life span of the fly coupled with additional genetic tools unique to this system will allow me to assess the impact of altered ACh release on behavior and synaptic physiology throughout the lifespan of the animal. In addition, I will test the hypothesis that an increase in cholinergic tone through overexpression of VAChT will improve age-related deficits in cognitive functions. In future aims, I will use the model system I propose here as a platform to identify small molecules capable of increasing ACh release. I believe that these agents could represent a new therapeutic strategy to improve the cognitive decline associated with decreased ACh neurotransmission in both normal and pathological aging.

 描述（由申请人提供）：我在未来五年的职业目标是获得R 01资金，在特拉华州州立大学的任期，并开发一个研究利基在使用果蝇阐明乙酰胆碱释放的分级变化对衰老过程中的生理和行为的作用。作为一个研究生与詹妮斯奥唐纳（美国）。作为亚拉巴马大学的博士后研究员和大卫克兰茨（加州大学洛杉矶分校）的博士后研究员，我接受了果蝇神经遗传学和与人类疾病相关的神经元过程研究的广泛培训。此外，Krantz博士是研究囊泡转运蛋白及其在神经传递中作用的专家，我在囊泡神经递质转运蛋白的功能方面积累了自己的专业知识。我最近的项目涉及使用果蝇转运蛋白来测试可能导致帕金森病（PD）易感性的基因-环境相互作用。特别是，我确定了囊泡单胺转运蛋白（VMAT）的过度表达可以通过将胞质多巴胺从其作用部位隔离来保护多巴胺神经元免受环境毒素的影响。为了建立VMAT的潜在神经保护作用，我进行了一项能够增加其功能的小分子筛选，并成功地鉴定了一类潜在的新型胺能药物。我将继续研究果蝇囊泡神经递质转运蛋白的神经遗传学在我的职业生涯的早期阶段作为一个独立的教师，但改变我的主要重点从VMAT到果蝇囊泡乙酰胆碱转运蛋白（VAChT）。在Krantz博士的出色指导下，我扩展了我的技能，包括细胞培养，行为分析，生化分离和药理学。我现在正在利用这个背景来确定VAChT在我自己的实验室中作为PI的生物学功能。我已经制定了一个严格的培训计划，这将有助于我实现我的职业过渡到衰老过程中胆碱能神经传递的研究。在K 01奖期间，我的培训的一个重要方面将是学习电生理学。我接受过Diane O 'Dowd的基本训练，她是成年果蝇脑电生理学专家。我学会了如何制备苍蝇的脑组织，并识别果蝇的胆碱能神经元。此外，我还收到了来自加州大学洛杉矶分校的Felix Schweizer关于一般电生理学概念和技术的输入。在这两位导师的指导下，我能够建立起连接神经元和记录动作电位的基础。我现在打算掌握这些基本技能，成为一名专家。施韦策博士和奥多德博士都表示愿意继续用他们的专业知识支持我。重要的是，特拉华州的梅丽莎哈灵顿博士提出要加强我的训练，并将担任我的动手初级生理学导师。她给了我充分的机会使用她的钻机，在她的指导下，我将继续磨练我的技能。她还为目前的申请提供了大量投入。我 我最近在特拉华州的NIH/COBRE神经科学项目中担任助理教授，建立了自己的实验室。我收到了32万美元的启动资金。这种支持为我实现目标提供了宝贵的资源/人员（见培训计划）。我的目标是成为一名使用电生理学研究ACh释放差异如何影响突触功能的专家。我将把这些技能与我目前的专业知识结合起来，以确定胆碱能突触功能的变化如何与衰老过程中的行为衰退相关。此外，我将很好地解决有关乙酰胆碱释放，行为和衰老的基本问题，具有明确的翻译意义。为了研究这些问题，我将使用 果蝇VAChT中的等位基因系列点突变体，该蛋白质包装和转运ACh以进行突触释放。这些等位基因的范围从轻度到重度，将使我能够确定乙酰胆碱释放的分级变化和乙酰胆碱介导的行为（如学习）的进行性缺陷之间的关系。果蝇的寿命很短，再加上这个系统特有的额外遗传工具，将使我能够评估改变ACh释放对动物整个生命周期的行为和突触生理学的影响。此外，我将测试的假设，增加胆碱能紧张通过过度表达VAChT将改善与年龄相关的认知功能的缺陷。在未来的目标中，我将使用我在这里提出的模型系统作为一个平台，以确定能够增加乙酰胆碱释放的小分子。我相信这些药物可以代表一种新的治疗策略，以改善与正常和病理性衰老中乙酰胆碱神经传递减少相关的认知能力下降。