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

 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）：我未来五年的职业目标是获得 R01 资助、确保在特拉华州立大学的终身教职，并开发利用果蝇的研究领域来阐明衰老过程中乙酰胆碱释放的分级变化对生理和行为的作用。作为 Janis O'Donnell（阿拉巴马大学）的研究生和 David Krantz（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方面的广泛培训。此外，与研究囊泡转运蛋白及其在神经传递中的作用的专家 Krantz 博士一起，我在囊泡神经递质转运蛋白的功能方面发展了自己的专业知识。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) 易感性的基因与环境相互作用。特别是，我确定囊泡单胺转运蛋白（VMAT）的过度表达可以通过隔离胞质多巴胺远离其作用位点来保护多巴胺神经元免受环境毒素的影响。为了发挥 VMAT 潜在的神经保护作用，我对能够增强其功能的小分子进行了筛选，并成功鉴定出了一类潜在的新型胺能药物。在我作为独立教师的职业生涯早期阶段，我将继续研究果蝇囊泡神经递质转运蛋白的神经遗传学，但将我的主要关注点从 VMAT 改为果蝇囊泡乙酰胆碱转运蛋白 (VAChT)。在 Krantz 博士的出色指导下，我扩展了自己的技能范围，包括细胞培养、行为分析、生化分离和药理学。现在，作为 PI，我正在自己的实验室中利用这一背景来确定 VAChT 的生物学功能。我制定了严格的培训计划，这将帮助我实现职业生涯过渡到衰老过程中胆碱能神经传递的研究。 K01 奖期间我训练的一个重要方面是学习电生理学。我接受了成年果蝇大脑电生理学专家黛安·奥多德 (Diane O'Dowd) 的基础培训。我学习了如何制备果蝇脑组织并识别果蝇胆碱能神经元。此外，我还从加州大学洛杉矶分校的 Felix Schweizer 那里获得了关于一般电生理学概念和技术的建议。在这两位导师的指导下，我能够建立修补神经元和记录动作电位的基础知识。我现在计划掌握这些基础技能并成为专家。两位博士。施韦泽和奥多德表示愿意继续用他们的专业知识支持我。重要的是，特拉华州立大学的梅丽莎·哈林顿博士主动提出加强我的培训，并将担任我的初级生理学导师。她让我可以完全使用她的装备，在她的指导下，我将继续磨练我的技能。她还为当前的申请提供了大量意见。我 最近，我在特拉华州立大学的 NIH/COBRE 神经科学项目中担任助理教授，创建了自己的实验室。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）。我的目标是成为利用电生理学研究乙酰胆碱释放差异如何影响突触功能的专家。我会将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下降相关。此外，我将能够很好地解决有关乙酰胆碱释放、行为和衰老的基本问题，这些问题具有明确的转化意义。为了研究这些问题，我将使用 果蝇 VAChT 中的一系列等位基因点突变体，VAChT 是包装和运输 ACh 以进行突触释放的蛋白质。这些等位基因的范围从轻微到严重，使我能够确定乙酰胆碱释放的分级变化与乙酰胆碱介导的行为（例如学习）的渐进性缺陷之间的关系。果蝇的寿命较短，加上该系统特有的额外遗传工具，将使我能够评估乙酰胆碱释放的改变对动物整个生命周期的行为和突触生理学的影响。此外，我将验证以下假设：通过 VAChT 过度表达增加胆碱能张力将改善与年龄相关的认知功能缺陷。在未来的目标中，我将使用我在这里提出的模型系统作为平台来识别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，以改善正常和病理衰老中与乙酰胆碱神经传递减少相关的认知能力下降。