Molecular Mechanism and Regulation of Asynchronous Release

异步释放的分子机制及调控

• 批准号: 10292969
• 负责人: Qiangjun Zhou
• 金额: $ 23.99万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10292969
• 关键词: Action Potentials; Architecture; Area; Atomic Resolution X-Ray Crystallography; Attention; Award; Binding; Biological Assay; Brain; Brain Stem; Collaborations; Communication; Complement; Complex; Cryoelectron Microscopy; Crystallization; Data; Development Plans; Doctor of Philosophy; Electrophysiology (science); Environment; Goals; Grant; Image; In Vitro; Investigation; Kinetics; Knockout Mice; Knowledge; Laboratories; Lead; Lipids; Measures; Mediating; Membrane; Membrane Proteins; Memory; Mental Depression; Mental disorders; Mentors; Mentorship; Methods; Molecular; Mutagenesis; Mutate; Neurons; Neurosciences; Pharmaceutical Preparations; Phase; Physiology; Play; Prevention; Proteins; Regulation; Research; Research Personnel; Role; Running; SNAP receptor; Schizophrenia; Scientific Advances and Accomplishments; Scientist; Sequence Analysis; Series; Slice; Spectrum Analysis; Structure; Synapses; Synaptic Vesicles; Synaptic plasticity; Technical Expertise; Techniques; Training; Training Programs; Vesicle; Work; base; career; career development; cellular imaging; design; learned behavior; medical schools; nanodisc technology; neural circuit; neurotransmission; neurotransmitter release; novel therapeutics; particle; relating to nervous system; sensor; single molecule; structural biology; synaptotagmin; tool

Project Summary/Abstract Neurotransmitter release is influenced by drugs, and certain mental disorders such as depression, but the details are unclear. It is mediated by the SNARE complex, complexin (Cpx), synaptotagmin (Syt), and other proteins. Asynchronous release is a mode of neurotransmitter release. It has received increasing attention as it may generate or compensate abnormal neural activities, and play critical role in pre-synaptic plasticity. While most neuronal communication relies upon synchronous release triggered by the Ca2+ sensors Syt1, 2 and 9 (collectively called “fast Syts”). Asynchronous release has a longer, variable delay after an action potential or series of action potentials, and it is mediated by the Ca2+ sensor Syt7. Fast Syts and Syt7 share a similar domain structure and a high degree of homology in their Ca2+-binding C2A and C2B domains. Recently I have identified the "primary" SNARE-Syt1 interface between Syt1 C2B domain and the SNARE complex, revealing the "primary" interface is specific for synchronous release. In contrast, Syt7 does not form a similar "primary" interface. These results raise the following questions: 1) how does Syt7's cooperate with SNAREs and Cpx to trigger "delayed" asynchronous release; 2) what is the difference between fast Syts and Syt7 that gives rise to the different modes of neurotransmitter release; 3) how is asynchronous release regulated by Syts and Ca2+. The overall goal of this proposal is the elucidation of the molecular mechanism and regulation of asynchronous release. The proposed study will examine how Syt7 mediates asynchronous release in cultured cortical neurons and brain slices from Syt1/7 double knockout mice with re-introduction of mutates designed based on sequence analysis and a newly solved crystal structure of SNARE-Cpx-Syt1 complex. Finally, I will reveal the interaction among Syt1/7, Cpx, SNARE complex and membrane, and investigate the regulation of asynchronous release by Syt1, Syt7 and Ca2+. Combined with my work on synchronous release, such results are expected to provide a better understanding of the roles of neurotransmitter release in neural activity and mental disorders, may lead to new therapeutics for the prevention and treatment of a variety of mental disorders. In addition, the results are also expected to vertically advance the understanding of pre-synaptic plasticity which is believed to be related to memory, learning, and behavior, as well as fundamentally advance the field of neuroscience. My long-term career goal is to lead a world-class laboratory in the forefront of synaptic physiology. I intend to use techniques in structural biology, single molecule spectroscopy and imaging, as well as key techniques in neuroscience as investigation tools. To complement my knowledge of structural biology acquired during my PhD, I will carry out the mentored phase of this Award as a postdoctoral Research Fellow in the laboratories of Drs. Axel Brunger and Thomas Südhof. I have designed an ambitious research career development plan to achieve my immediate goals for the mentored period: 1) expand my advanced scientific and technical knowledge, and 2) prepare my transition to independence. Under the mentorship of Drs. Brunger and Südhof, I will follow a structured training program to enhance my professional abilities to establish and run my own laboratory. Stanford Medical School will provide me with ideal environment to fully benefit from this Award and become a successful independent scientist.

项目概要/摘要 神经递质的释放受到药物和某些精神疾病（例如抑郁症）的影响，但细节是 不清楚。它由 SNARE 复合体、复合蛋白 (Cpx)、突触结合蛋白 (Syt) 和其他蛋白质介导。异步 释放是神经递质释放的一种模式。它受到越来越多的关注，因为它可能产生或补偿 异常的神经活动，并在突触前可塑性中发挥关键作用。虽然大多数神经元通讯依赖于 由 Ca2+ 传感器 Syt1、2 和 9（统称为“快速 Syts”）触发同步释放。异步 在一个动作电位或一系列动作电位之后，释放有较长的、可变的延迟，并且由 Ca2+ 介导 传感器 Syt7。 Fast Syts 和 Syt7 具有相似的结构域结构，并且它们的 Ca2+ 结合 C2A 具有高度的同源性 和 C2B 领域。最近我已经确定了 Syt1 C2B 域和 SNARE复杂，揭示了“主要”接口是专门用于同步释放的。相比之下，Syt7 不形成 类似的“主”界面。这些结果提出了以下问题：1）Syt7 如何与 SNARE 合作以及 Cpx触发“延迟”异步释放； 2）快速Syts和Syt7之间的区别是什么导致了 不同的神经递质释放模式； 3）Syts和Ca2+如何调节异步释放。总体目标 该提案的目的是阐明异步释放的分子机制和调控。拟议的 研究将检查 Syt7 如何介导培养的皮质神经元和 Syt1/7 脑切片的异步释放 重新引入基于序列分析和新解决的晶体设计的突变体的双敲除小鼠 SNARE-Cpx-Syt1 复合物的结构。最后，我将揭示 Syt1/7、Cpx、SNARE 复合体和 膜，并研究 Syt1、Syt7 和 Ca2+ 对异步释放的调节。结合我的工作 同步释放，这样的结果有望更好地理解神经递质释放在 神经活动和精神障碍，可能会带来预防和治疗各种精神疾病的新疗法 失调。此外，该结果也有望纵向推进对突触前可塑性的理解 它被认为与记忆、学习和行为有关，并从根本上推进了这一领域 神经科学。我的长期职业目标是领导突触生理学前沿的世界一流实验室。我打算 使用结构生物学、单分子光谱学和成像技术以及关键技术 神经科学作为研究工具。为了补充我在博士学位期间获得的结构生物学知识，我将 作为博士后研究员在博士实验室中开展该奖项的指导阶段。阿克塞尔·布伦格 和托马斯·苏德霍夫。我设计了一个雄心勃勃的研究职业发展计划，以实现我的近期目标 指导期间：1）扩展我的先进科学和技术知识，2）准备我的过渡 独立。在博士的指导下。 Brunger 和 Südhof，我将遵循结构化的培训计划来提高 我建立和经营自己的实验室的专业能力。斯坦福医学院将为我提供理想的 充分受益于该奖项并成为一名成功的独立科学家的环境。

项目成果

Glia-derived secretory fatty acid binding protein Obp44a regulates lipid storage and efflux in the developing Drosophila brain.

胶质细胞衍生的分泌性脂肪酸结合蛋白 Obp44a 调节发育中的果蝇大脑中的脂质储存和流出。

• DOI: 10.1101/2024.04.10.588417
• 发表时间: 2024
• 期刊: bioRxiv : the preprint server for biology
• 作者: Yin,Jun;Chen,Hsueh-Ling;Grigsby-Brown,Anna;He,Yi;Cotten,MyriamL;Short,Jacob;Dermady,Aidan;Lei,Jingce;Gibbs,Mary;Cheng,EthanS;Zhang,Dean;Long,Caixia;Xu,Lele;Zhong,Tiffany;Abzalimov,Rinat;Haider,Mariam;Sun,Rong;He,Ye;Zh
• 通讯作者: Zh