Imaging exocytosis via a combined fluorescence and amperometry approach

通过荧光和电流分析相结合的方法对胞吐作用进行成像

• 批准号: 2203359
• 负责人: Timothy Glass
• 金额: $ 56.02万
• 依托单位: University of Missouri-Columbia
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2203359&HistoricalAwards=false
• 关键词: Imaging; exocytosis; via; combined; fluorescence

With the support of the Chemistry of Life Processes (CLP) Program in the Division of Chemistry, Professors Timothy Glass and Kevin Gillis of the University of Missouri are studying how neurotransmitters are released into synapses. Neurotransmitters are critical to the regulation of the nervous system and control a number of functions such as learning, memory, sleep, and movement. Understanding the machinery of synaptic release, and the chemical activity of neurotransmitters is vital to understanding both normal and atypical neuronal processes. Furthermore, understanding the basic mechanisms of synaptic vesicle fusion and transmitter release via exocytosis is of broad significance because it will not only aid in the development of therapies for diseases where release of neurotransmitters is compromised, but it will also advance our understanding of FDA-approved treatments that modulate transmitter release, such as botulinum toxin A and B. As part of the broader impacts of this work, a new robotics camp for underserved high school students will be held. It has been demonstrated that activities such as these camps enhances the likelihood that participants pursue STEM (science, technology, engineering and mathematics) coursework during their secondary education.This project involves the preparation and evaluation of fluorescent chemical sensors for catecholamines with a view toward the fluorescent detection of neurotransmitters. These sensors are related to the NeuroSensor class of probes developed in PI Glass’s lab, which have already been used to detect norepinephrine in isolated chromaffin cells. Sensors will be developed that produce fluorescence enhancements upon exocytosis. The sensors will be used in combination with novel transparent electrodes developed in PI Gillis’ lab to study mechanisms of exocytosis via fluorescence imaging in combination with amperometric measurements. These combination experiments will measure both release and retention of catecholamine from individual vesicles to test the hypothesis that the amount of catecholamine released from a vesicle is modulated by the stimulus intensity. This project is expected to result in the development and application of a new set of important chemical biology tools for the study of neurochemistry with potentially broad scientific impact.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在化学系生命过程化学（CLP）项目的支持下，密苏里大学的Timothy Glass教授和Kevin Gillis教授正在研究神经递质是如何释放到突触中的。神经递质对神经系统的调节至关重要，并控制许多功能，如学习、记忆、睡眠和运动。了解突触释放机制和神经递质的化学活性对于理解正常和非典型神经元过程至关重要。此外，了解突触囊泡融合和通过胞外作用释放递质的基本机制具有广泛的意义，因为它不仅有助于开发神经递质释放受损的疾病的治疗方法，而且还将促进我们对fda批准的调节递质释放的治疗方法的理解，如肉毒杆菌毒素A和b。一个新的机器人训练营将为缺乏服务的高中生举办。事实证明，这些营地之类的活动提高了参与者在中学教育期间学习STEM（科学、技术、工程和数学）课程的可能性。本项目涉及儿茶酚胺荧光化学传感器的制备和评价，以期对神经递质进行荧光检测。这些传感器与PI Glass实验室开发的神经传感器（NeuroSensor）探针有关，后者已被用于检测分离的染色质细胞中的去甲肾上腺素。在胞吐过程中产生荧光增强的传感器将被开发出来。这些传感器将与PI Gillis实验室开发的新型透明电极结合使用，通过荧光成像和安培测量相结合来研究胞吐的机制。这些组合实验将测量单个囊泡中儿茶酚胺的释放和保留，以验证从囊泡中释放的儿茶酚胺的量受刺激强度调节的假设。该项目有望开发和应用一套新的重要化学生物学工具，用于神经化学研究，具有潜在的广泛科学影响。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。