Acetylcholinergic Neurotransmission During Aging

衰老过程中的乙酰胆碱能神经传递

• 批准号: 9130071
• 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/9130071
• 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; Career Mobility; 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; Foundational Skills; Fractionation; Functional disorder; Funding; Future; Genes; Genetic; Genetic Techniques; Goals; Health; 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 therapeutic intervention; overexpression; pathological aging; presynaptic; professor; programs; research study; response; skills; small molecule; synaptic function; tool; trafficking; vesicular monoamine transporter; Acetylcholine; Action Potentials; Address; Adult; Affect; Age; Aging; Alabama; Alleles; Alzheimer' s Disease; Animals; Behavior; Behavioral; Behavioral Mechanisms; Biochemical; Biological Models; Biological Process; Brain; Career Mobility; 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; Foundational Skills; Fractionation; Functional disorder; Funding; Future; Genes; Genetic; Genetic Techniques; Goals; Health; 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 therapeutic intervention; overexpression; pathological aging; presynaptic; professor; programs; 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（由适用提供）：我未来五年内的职业目标是获得R01资金，在特拉华州立大学获得安全任期，并在使用果蝇来阐明乙烯胆碱释放对生理学和行为的渐变作用方面开发了一个研究利基市场。作为Janis O'Donnell（美国阿拉巴马州）的研究生和David Krantz（UCLA）的博士后研究员，我接受了果蝇神经遗传学的广泛培训以及研究与人类疾病有关的神经过程。此外，与囊泡转运蛋白研究专家Krantz博士及其在神经传递中的作用，我已经开发了自己的专家在囊泡神经递质转运蛋白的功能方面。我最近的项目涉及使用果蝇转运蛋白测试可能有助于帕金森氏病（PD）易感的基因环境相互作用。特别是，我确定囊泡单胺转运蛋白（VMAT）的过表达可以通过隔离胞质多巴胺从其作用部位隔离，可以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，我为能够提高其功能的小分子进行了筛选，我成功地确定了潜在的新类AMINEGIC剂。我将继续研究我职业生涯的早期果蝇神经递质转运蛋白的神经遗传学作为独立的教师，但将我的主要重点从VMAT转变为果蝇乙酰乙酰胆碱胆碱转运蛋白（VACHT）。在Krantz博士的出色心态下，我扩大了技能的曲目，包括细胞培养，行为分析，生化分馏和药理学。现在，我正在利用此背景来确定我自己的实验室中VACHT的生物学功能作为PI。我已经制定了一项严格的培训计划，这将帮助我实现自己的职业过渡到对衰老期间胆碱能神经传递的研究。我在K01奖项期间培训的一个重要方面是学习电生理学。我接受了成人果蝇脑电生理学专家黛安·奥多德（Diane O'Dowd）的基础培训。我学会了如何准备蝇脑组织并鉴定果蝇胆碱能神经元。此外，我收到了UCLA Felix Schweizer的一般电生理概念和技术的输入。在这两个导师的情况下，我能够建立在神经元和记录动作电位上的补丁基础。我现在计划掌握这些基本技能并成为专家。两个博士。 Schweizer和O'Dowd提议继续以他们的专业知识来支持我。重要的是，特拉华州立大学的梅利莎·哈灵顿（Melissa Harrington）博士提出，旨在增强我的培训，并将作为我动手的主要生理学精神。她给了我完全进入钻机的机会，在她的心态下，我将继续磨练自己的技能。她还向当前申请提供了大量投入。我 最近，我在特拉华州立大学的NIH/Cobre神经科学计划的助理教授开始了我自己的实验室。我收到了32万美元的启动套餐。这种支持为我提供了宝贵的资源/人员来实现我的目标（请参阅培训计划）。我的目标是成为使用电生理学的专家，以研究ACH释放效应突触功能的差异。我将把这种技能与当前的专业知识相结合，以确定胆碱能突触功能的变化如何与衰老期间的行为下降相关。此外，我将有充分的态度解决有关ACH释放，行为和衰老的基本问题，这些问题具有明显的翻译意义。为了研究这些问题，我将使用 果蝇中的等位基因突变体系列突变体是包装和运输ACH以进行突触释放的蛋白质。这些等位基因范围从轻度到重度，可以确定乙酰胆碱释放的分级变化与渐进式的分级变化之间的关系，定义了乙酰胆碱介导的行为（例如学习）。苍蝇的短寿命以及该系统独有的其他遗传工具将使我能够评估ACH释放对动物整个生命周期中行为和突触生理学的影响。此外，我将检验以下假设：通过vacht的过表达胆碱能张力的增加将改善与年龄相关的认知功能的缺陷。在将来的目标中，我将在此处使用模型系统作为一个平台，以识别能够增加ACH释放的小分子。我认为，这些药物可以代表一种新的理论策略，以改善正常和病理衰老中ACH神经传递降低相关的认知下降。