The Impact of Beta- and Gamma-synucleins on Alpha-synuclein's Synaptic Function

β 和 γ 突触核蛋白对 α 突触核蛋白突触功能的影响

• 批准号: 10830522
• 负责人: Jacqueline Burre
• 金额: $ 3.66万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10830522
• 关键词: Affect; Age; Alzheimer' s disease related dementia; Amyotrophic Lateral Sclerosis; Animals; Area; Basic Science; Binding; Biochemical; Biophysics; Brain; Cells; Complex; Dementia; Development; Disease; Electrophysiology (science); Exocytosis; Fostering; Functional disorder; Gamma synuclein; Gaucher Disease; Hippocampus; Knockout Mice; Lewy Body Dementia; Link; Medical; Mission; Modeling; Molecular; Molecular Chaperones; Mus; Neurodegenerative Disorders; Neurons; Parkinson Disease; Pathologic; Pathology; Physiological; Play; Public Health; Recombinant Proteins; Research; Role; SNAP receptor; Slice; Synapses; Synaptic Vesicles; Testing; Therapeutic Intervention; Translational Research; United States National Institutes of Health; VAMP-2; Work; alpha synuclein; innovation; insight; interdisciplinary approach; mouse model; nervous system disorder; neurotransmitter release; novel; novel therapeutic intervention; phosphoneuroprotein 14; prevent; synaptic function; synuclein; synucleinopathy; virtual

PROJECT SUMMARY/ABSTRACT αSynuclein (αSyn) plays an important role at the synapse, to maintain neurotransmitter release via clustering synaptic vesicles (SV) and chaperoning SNARE-complex assembly. Aggregation of αSyn is a key pathological feature in multiple age-driven neurodegenerative diseases such as Parkinson’s disease (PD) and Alzheimer’s disease related dementias (ADRD) such as Lewy body dementia. Despite the involvement of βSyn and γSyn in synucleinopathies including Lewy body dementia, Gaucher’s disease, and PD, virtually nothing is known about their physiological functions in the brain. Understanding their function is the first step to finding out how their dysfunction causes PD and Alzheimer’s disease related dementias including Lewy body dementia, and how these diseases can be prevented or delayed. The objective here is to determine the effects of βSyn and γSyn on αSyn’s synaptic function. The central hypothesis is that interaction of βSyn and γSyn with αSyn causes a reduction in αSyn’s activity, leading to reduced SV clusters and SNARE-complex assembly, and altered neu- ronal activity. This hypothesis will be tested in 3 specific aims: 1) Assess the effect of βSyn and γSyn on SNARE-complex assembly; 2) Assess the effect of βSyn and γSyn on SV clustering; and 3) Determine the im- plications of synuclein interactions on SV cycling. Under aim 1, SNARE-complex assembly will be quantified in select brain areas and neurons from mice lacking βSyn and/or γSyn, in heterologous cells and using recombi- nant proteins. Under aim 2, synaptobrevin-2 binding and multimerization of αSyn, and SV pools will be quanti- fied in mice lacking βSyn and/or γSyn and using recombinant proteins. Under aim 3, SV exocytosis and cycling will be quantified in hippocampal brain slices and in neurons from mice lacking βSyn and/or γSyn with or with- out lentivirally increasing βSyn or γSyn levels. This research is innovative because it (1) tests the novel hypoth- esis that βSyn and γSyn affect the synaptic function of αSyn, (2) uses a multidisciplinary approach combining biophysical, biochemical, electrophysiological and whole animal approaches, and (3) analyzes new mouse models lacking βSyn and/or γSyn that were generated from αβγSyn triple knockout mice, that not only enable a direct comparison of the synucleins but may serve as models for synucleinopathies and Alzheimer’s disease related dementias including Lewy body dementia. Our work is significant because it (1) will clarify the im- portance of βSyn and γSyn for neuronal function, (2) will provide new insights into the molecular mechanism underlying SV binding of αSyn, (3) may uncover the contributions of βSyn and γSyn to synucleinopathies and Alzheimer’s disease related dementias including Lewy body dementia, and (4) has translational importance for the targeted development of new treatment strategies for the above-mentioned age-driven dementias aimed at targeting all synucleins instead of focusing solely on αSyn. With synuclein pathology and co-pathologies com- mon to PD and Alzheimer’s disease related dementias including Lewy body dementia, our study has the poten- tial to contribute a mechanistic understanding of the role of the three synucleins in these synucleinopathies.

项目总结/摘要 α-突触核蛋白（α-Syn）在突触中起重要作用，通过聚集维持神经递质的释放 突触囊泡（SV）和伴侣SNARE复合物组装。α-Syn的聚集是一种关键的病理性 在多种年龄驱动的神经退行性疾病，如帕金森病（PD）和阿尔茨海默氏症的特征 疾病相关性痴呆（ADRD），如路易体痴呆。尽管βSyn和γSyn参与了 包括路易体痴呆、戈谢病和PD在内的共核蛋白病，实际上对其一无所知 它们在大脑中的生理功能了解它们的功能是了解它们如何 功能障碍导致PD和阿尔茨海默病相关痴呆，包括路易体痴呆，以及如何 这些疾病是可以预防或延缓的。这里的目的是确定βSyn和γSyn的影响 α-突触的突触功能中心假设是βSyn和γSyn与αSyn的相互作用导致了α Syn与β Syn的相互作用。 αSyn活性降低，导致SV簇和SNARE复合物组装减少，并改变neu- ronal活动该假设将在3个具体目标中进行检验：1）评估βSyn和γSyn对 SNARE-复合物组装; 2）评估βSyn和γSyn对SV聚类的影响;以及3）确定非线性函数。 突触核蛋白相互作用对SV循环的影响。根据目标1，SNARE复合物组装将在 选择来自缺乏βSyn和/或γSyn的小鼠的脑区域和神经元，在异源细胞中并使用重组- nant蛋白在目标2下，将定量分析突触泡蛋白-2结合和αSyn和SV池的多聚化。 在缺乏βSyn和/或γSyn并使用重组蛋白的小鼠中进行。在目标3下，SV胞吐和循环 将在海马脑切片和缺乏βSyn和/或γSyn的小鼠的神经元中定量， 慢病毒增加βSyn或γSyn水平。这项研究是创新的，因为它（1）测试了新的假设- βSyn和γSyn影响αSyn的突触功能，（2）采用多学科结合的方法， 生物物理学、生物化学、电生理学和整体动物方法，以及（3）分析新的小鼠 从αβγSyn三重敲除小鼠产生的缺乏βSyn和/或γSyn的模型， 突触核蛋白的直接比较，但可以作为突触核蛋白病和阿尔茨海默病的模型 相关痴呆症包括路易体痴呆症我们的工作是有意义的，因为它（1）将澄清的im- βSyn和γSyn在神经元功能中的重要性，（2）将为神经元功能的分子机制提供新的见解 αSyn的潜在SV结合，（3）可能揭示βSyn和γSyn对突触核蛋白病的贡献， 阿尔茨海默病相关痴呆，包括路易体痴呆，和（4）具有翻译重要性， 有针对性地为上述年龄驱动的痴呆症制定新的治疗策略， 靶向所有的突触核蛋白而不是只关注αSyn。与突触核蛋白病理学和共同病理学- 帕金森病和阿尔茨海默病相关痴呆包括路易体痴呆，我们的研究有可能 有助于对这三种突触核蛋白在这些突触核蛋白病中的作用机制的理解。