The role of beta-synuclein in synaptic vesicle release

β-突触核蛋白在突触小泡释放中的作用

• 批准号: 10536035
• 负责人: Lauren Komer
• 金额: $ 4.68万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-15 至 2025-06-14
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10536035
• 关键词: Address; Affect; Area; Basic Science; Binding; Biochemical; Biological; Biological Assay; Brain; Brain region; Cells; Complex; Data; Development; Diffuse Lewy Body Disease; Disease; Electron Microscopy; Endocytosis; Exhibits; Exocytosis; Fluorescent Probes; Fostering; Functional disorder; Gaucher Disease; Human; Knock-out; Knockout Mice; Knowledge; Lead; Lewy Bodies; Lewy Body Dementia; Link; Measures; Mediating; Medical; Membrane; Methods; Microelectrodes; Mission; Molecular; Molecular Chaperones; Multiple System Atrophy; Mus; Nervous system structure; Neurodegenerative Disorders; Neurons; PHluorin; Parkinson Disease; Pathogenicity; Pathologic; Pathology; Physiological; Play; Proteins; Public Health; Publications; Publishing; Research; Role; SNAP receptor; Synapses; Synaptic Vesicles; Synaptophysin; Testing; Therapeutic; Therapeutic Intervention; Translational Research; United States National Institutes of Health; Vesicle; Western Blotting; Wild Type Mouse; Work; alpha synuclein; base; brain tissue; combat; experimental study; innovation; insight; interdisciplinary approach; lentivirally transduced; nervous system disorder; neurotransmitter release; novel; novel strategies; novel therapeutic intervention; phosphoneuroprotein 14; protein function; single molecule; synuclein; synucleinopathy; vesicular release

PROJECT SUMMARY/ABSTRACT α-Synuclein (αSyn) and β-synuclein (βSyn) are abundantly expressed proteins found throughout the vertebrate nervous system. αSyn aggregation is pathologically associated with Parkinson’s disease (PD), multiple system atrophy, Lewy body dementia (LBD), and many other synucleinopathies. βSyn is known to be involved in neu- rodegenerative diseases such as LBD, diffuse Lewy body disease, Gaucher’s disease, and PD, but almost nothing is known about its function physiologically. Preliminary data demonstrate that direct interaction of βSyn with αSyn results in reduced synaptic vesicle binding of αSyn, potentially affecting αSyn function in the brain. The objective of this proposal is to delineate the functional consequences of the interaction of βSyn with αSyn for a neuron. The central hypothesis of this proposal, based on strong preliminary data, is that βSyn reduces αSyn’s ability to support SNARE-complex assembly, synaptic vesicle clustering and thereby neuro- transmitter release. The rationale for these studies is that understanding the effects of βSyn on αSyn function may lead to a novel understanding of synuclein function in the brain, and may lie the groundwork for the devel- opment of novel therapeutic strategies for combating synucleinopathies by modulating βSyn instead of αSyn directly. Guided by preliminary data, this hypothesis will be tested in two specific aims: Aim 1) Determine the effect of βSyn on αSyn-mediated SNARE-complex assembly and synaptic vesicle clustering, and Aim 2) Deter- mine how βSyn affects neuronal activity. In the first aim, SNARE-complex levels and synaptic vesicle cluster- ing will be quantified as a function of varied αSyn/βSyn ratios, in P40 WT and βSyn KO mouse brain tissue punches of select brain regions, and in primary neurons with varied βSyn levels. In the second aim, changes to spontaneous and evoked neurotransmitter release in βSyn KO neuronal cultures lentivirally expressing in- creasing amounts of βSyn will be measured, using a microelectrode array and synaptopHluorin experiments. We expect βSyn to reduce αSyn-mediated SNARE-complex assembly and synaptic vesicle clustering. We also expect βSyn to reduce the effect of αSyn on synaptic vesicle release and cycling, due to restraining αSyn away from synaptic vesicles and towards a more soluble and functionally inactive pool. This research is significant because understanding the molecular mechanism of how βSyn functions broadens our understanding of the function of all synucleins, and may provide insight into αSyn dysfunction in synucleinopathies. Previous work focused on eliminating αSyn from neurons was has shown to be detrimental. Altering βSyn instead of aSyn directly may provide a potential therapeutic avenue, but much research remains to be done. This research is innovative because of (1) its novel hypothesis that βSyn affects αSyn function, (2) the proposed study directly addresses the lack of knowledge into the molecular function of βSyn and its effect on αSyn, which could open up avenues of research for new synucleinopathies therapeutics, and (3) its multidisciplinary approach combin- ing biochemical and cell biological approaches to gain insight into the physiological function of βSyn.

项目摘要/摘要 α-突触核蛋白（αSyn）和β-突触核蛋白（βSyn）在整个脊椎动物中绝对表达蛋白质 神经系统。 αSyn聚集在病理上与帕金森氏病（PD），多系统相关 萎缩，刘易体内痴呆（LBD）和许多其他突触核酸病变。已知βSyn参与neu- LBD，弥漫性Lewy身体疾病，Gaucher病和PD等牛仔疾病，但几乎 关于其物理功能一无所知。初步数据表明βSyn的直接相互作用 随着αSyn导致αSyn的合成囊泡结合降低，可能影响大脑中的αSyn功能。 该建议的目的是描述βSyn与αSyn相互作用的功能后果 用于神经元。该提案的中心假设基于强大的初步数据，是βSyn降低了 αSyn支持圈套复合组件，突触囊泡聚类的能力，从而 发射器释放。这些研究的理由是了解βSyn对αSyn功能的影响 可能会导致对大脑突触核蛋白功能的新了解，并可能为Devel- 通过调节βSyn而不是αSyn来对抗突触核虫病的新型热策略的选择 直接地。在初步数据的指导下，该假设将以两个具体的目的进行检验：目标1）确定 βSyn对αSyn介导的圈子复合组装和突触囊泡聚类的影响，而目标2） 我的βSyn如何影响神经元活性。在第一个目标中，圈套复合物水平和突触囊泡簇 - 在p40 wt和βSynKO小鼠脑组织中，将量化为各种αSyn/βSyn比的函数 精选的大脑区域的打孔，在具有不同βSyn水平的原发性神经元中。在第二个目标中，变更 βSynKO神经元培养物中的赞助和诱发神经递质释放 使用微电极阵列和突触荧光素实验，将测量折痕量的βSyn。 我们预计βSyn将减少αSyn介导的圈套复合体和突触囊泡聚类。我们也是 预计βsyn会因限制αSyn而降低αSyn对合成囊泡释放和循环的影响 从突触蔬菜到更固体和功能性的池。这项研究很重要 因为了解βSyn的分子机制如何扩大我们对 所有突触核蛋白的功能，并可能提供有关突触核蛋白病中αSyn功能障碍的见解。以前的工作 已经确定了针对从神经元中消除αSyn的。改变βSyn而不是ASYN 直接可以提供潜在的治疗途径，但仍有许多研究要做。这项研究是 创新性是因为（1）βSyn影响αSyn功能的新型假设，（2）拟议的研究直接 解决了对βSyn分子功能的缺乏及其对αSyn的影响的知识，这可能打开 增加新的突触核病治疗研究的途径，以及（3）其多学科方法结合了 生化和细胞生物学方法可深入了解βSyn的身体功能。