Role of SNARE Interactions in Central Synapse Function

SNARE 相互作用在中枢突触功能中的作用

• 批准号: 7325717
• 负责人: Ege T Kavalali
• 金额: $ 26.63万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-12-01 至 2009-07-07
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7325717
• 关键词: Action Potentials; Address; Amino Acids; Automobile Driving; Binding; Biological; Birth; Cell membrane; Cells; Chromosome Pairing; Cleaved cell; Complex; Dependence; Disease; Distant; Dyes; Endocytosis; Epilepsy; Eukaryota; Eukaryotic Cell; Exhibits; Exocytosis; Genetic; Hippocampus (Brain); Image; In Vitro; Investigation; Knock-out; Knockout Mice; Measures; Mediating; Membrane; Membrane Fusion; Membrane Proteins; Mental Retardation; Molecular; Mus; Mutate; N-terminal; Optics; Pathway interactions; Phenotype; Process; Property; Proteins; Rate; Reaction; Recycling; Research; Residual state; Resolution; Retrieval; Role; SNAP receptor; Schizophrenia; Signal Transduction; Specificity; Sucrose; Synapses; Synaptic Transmission; Synaptic Vesicles; System; Techniques; Testing; Thinking; Time; Toxin; Transfection; Transmembrane Domain; VAMP-2; Vesicle; analog; base; cellubrevin; design; executive function; insight; interest; mutant; nervous system disorder; neurotransmission; neurotransmitter release; novel therapeutics; protein protein interaction; rabphilin; receptor; receptor function; research study; response; soluble NSF attachment protein; synaptic function; syntaxin; syntaxin 1; trafficking; transmission process; vesicle-associated membrane protein

DESCRIPTION (provided by applicant): Synaptobrevin2/VAMP2 (syb2) is a synaptic vesicle associated protein critical for normal synaptic transmission in central synapses. Its interaction with plasma membrane associated proteins syntaxin1 and SNAP-25 is thought to be an important step in synaptic vesicle fusion. These proteins are called SNAREs (acronym for soluble NSF attachment protein receptors). Despite extensive progress on characterization of molecular interactions between SNAREs, their precise role in synaptic transmission remains elusive. Knockout mice lacking syb2 exhibit a lethal phenotype and die at birth. In preliminary experiments, analysis of hippocampal cultures extracted from these mice revealed a persistent form of regulated exocytosis and synaptic vesicle recycling. This result supports a role for SNAREs in regulating the rate and Ca2+-dependence of neurotransmission rather than an executive function in driving fusion. The main objective of this project is to test the hypotheses that emerge from this initial analysis of syb2 deficient mice and elucidate the role of syb2 and its interactions with other SNAREs in synaptic vesicle exocytosis and recycling in mammalian central synapses. 1. To determine the molecular basis for synaptic vesicle fusion and recycling observed in syb2 deficient synapses, the involvement of other SNAREs or vesicle membrane associated proteins in the remaining neurotransmission will be tested. 2. The role of SNARE interactions in vesicle retrieval and recycling will be investigated using styryl dye imaging. 3. To define structural requirements for syb2 function, wild type and mutant syb2 constructs will be introduced to hippocampal cultures obtained from knockout mice to study in vitro rescue. Information attained from this study will also provide new insight to the disease process in psychiatric and neurological disorder such as mental retardation, schizophrenia and epilepsy leading to design of new therapeutic strategies.

描述（由申请人提供）： Synaptobrevin2/VAMP2 (syb2) 是一种突触小泡相关蛋白，对于中枢突触的正常突触传递至关重要。它与质膜相关蛋白 Syntaxin1 和 SNAP-25 的相互作用被认为是突触小泡融合的重要步骤。这些蛋白质被称为 SNARE（可溶性 NSF 附着蛋白受体的缩写）。尽管在 SNARE 之间分子相互作用的表征方面取得了广泛进展，但它们在突触传递中的确切作用仍然难以捉摸。缺乏syb2的基因敲除小鼠表现出致命表型并在出生时死亡。在初步实验中，对从这些小鼠中提取的海马培养物的分析揭示了一种持续形式的受调节的胞吐作用和突触小泡再循环。这一结果支持了 SNARE 在调节神经传递速率和 Ca2+ 依赖性方面的作用，而不是在驱动融合中的执行功能。该项目的主要目的是测试对 syb2 缺陷小鼠的初步分析中提出的假设，并阐明 syb2 的作用及其与其他 SNARE 在哺乳动物中枢突触的突触小泡胞吐作用和再循环中的相互作用。 1. 为了确定在 syb2 缺陷突触中观察到的突触小泡融合和再循环的分子基础，将测试其他 SNARE 或囊泡膜相关蛋白在剩余神经传递中的参与情况。 2. 将使用苯乙烯基染料成像研究 SNARE 相互作用在囊泡回收和再循环中的作用。 3.为了确定syb2功能的结构要求，将野生型和突变型syb2构建体引入从敲除小鼠获得的海马培养物中以研究体外拯救。从这项研究中获得的信息还将为精神障碍、精神分裂症和癫痫等精神和神经疾病的疾病过程提供新的见解，从而设计新的治疗策略。