Investigating CaMKII regulation of extracellular vesicle trafficking to promote synaptic plasticity

研究 CaMKII 对细胞外囊泡运输的调节以促进突触可塑性

• 批准号: 10425693
• 负责人: Matthew F. Pescosolido
• 金额: $ 6.36万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2023-03-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10425693
• 关键词: 3-Dimensional; Affect; Alzheimer' s Disease; Biogenesis; Biological Process; Biology; Brain; Ca(2+)-Calmodulin Dependent Protein Kinase; Cell Communication; Cell model; Cells; Clathrin; Communication; Complex; Data; Drosophila genus; Early Endosome; Electrophysiology (science); Endosomes; Fostering; Genetic; Goals; Growth; Image; Image Analysis; In Vitro; Light; Link; Measurement; Mediating; Membrane; Methods; Microscopy; Modeling; Molecular; Multivesicular Body; Nervous system structure; Neurodegenerative Disorders; Neuroglia; Neuromuscular Junction; Neurons; Oranges; Parkinson Disease; Pathologic Processes; Pathway interactions; Pharmacology; Phosphotransferases; Play; Population; Presynaptic Terminals; Process; Proteins; Recycling; Regulation; Research; Resolution; Role; Signal Transduction; Sorting - Cell Movement; Synapses; Synaptic Vesicles; Synaptic plasticity; System; Testing; Training; Vesicle; calmodulin-dependent protein kinase II; dynactin; experimental study; extracellular vesicles; functional plasticity; genetic approach; imaging genetics; in vivo Model; insight; intercellular communication; late endosome; mutant; nervous system disorder; neuronal cell body; neurotransmitter release; postsynaptic; presynaptic; presynaptic neurons; protein aggregation; quantitative imaging; synaptotagmin IV; trafficking; vesicular release

PROJECT SUMMARY Extracellular vesicles (EVs) are small membrane-bound compartments that exchange materials between cells, and play an important role in cell communication in the nervous system. Neuronal EVs mediate activity- dependent synaptic plasticity, as well as the spread and clearance of toxic protein aggregates associated with neurodegenerative disorders such as Alzheimer’s Disease and Parkinson’s Disease. EV biogenesis and secretion occurs via endosomal trafficking, but the mechanisms by which neuronal activity regulates these cell biological processes have not been well characterized. By identifying activity-dependent mechanisms influencing neuronal EV dynamics, we will gain greater insight into basic and pathological processes in the nervous system. The majority of EV research utilizes in vitro non-neuronal cell models to isolate heterogenous EV populations, making direct measurements of neuronal EV biogenesis and trafficking mechanisms challenging. The Drosophila neuromuscular junction (NMJ) serves as a powerful in vivo model that is highly amenable to manipulating and visualizing EV cargo. My preliminary findings in this system suggest that Calcium/calmodulin-dependent protein kinase II (CaMKII), a key synaptic protein involved in structural and functional forms of synaptic plasticity, plays a role in EV trafficking. Specifically, CaMKII null mutants showed significantly decreased levels of the EV cargo Syt4 in both EV precursors in the presynaptic neuron and postsynaptically secreted EVs. Syt4 modulates NMJ synaptic growth and neurotransmitter release in an activity-dependent manner, leading to my hypothesis that CaMKII regulates Syt4 EV signaling to promote structural and functional plasticity. In Aim 1, I will ask if CaMKII controls EV cargo levels in an endocytic trafficking pathway that we previously identified to mediate EV cargo trafficking. In the second aim, I will elucidate the relationship between CaMKII and Syt4 and test if they act in a shared pathway to promote structural and functional plasticity. In Aim 3, I will isolate the specific CaMKII function(s) involved in EV cargo and trafficking regulation. These aims will provide training in advanced imaging, quantitative image analysis, Drosophila genetics, and electrophysiology. The information obtained from this project will provide new insight into how CaMKII regulates EV trafficking to facilitate synaptic plasticity, and identify new activity-dependent mechanisms regulating EVs in the nervous system.

项目概要 细胞外囊泡 (EV) 是小的膜结合区室，可在细胞之间交换物质， 并在神经系统的细胞通讯中发挥重要作用 - 依赖的突触可塑性，以及与相关的有毒蛋白质聚集体的传播和清除 神经退行性疾病，例如阿尔茨海默病和帕金森病，以及 EV 生物发生和。 分泌通过内体运输发生，但神经活动调节这些细胞的机制 生物过程尚未通过识别活动依赖的影响机制得到很好的表征。 神经元 EV 动力学，我们将更深入地了解神经系统的基本和病理过程。 大多数 EV 研究利用体外非神经元细胞模型来分离异质 EV 群体， 使得直接测量果蝇神经元 EV 生物发生和运输机制具有挑战性。 神经肌肉接头（NMJ）作为一种强大的体内模型，非常适合操纵和 我在该系统中的初步发现表明钙/钙调蛋白依赖性蛋白 激酶 II (CaMKII) 是一种关键的突触蛋白，参与突触可塑性的结构和功能形式，在 具体来说，CaMKII 无效突变体显示 EV 货物水平显着降低。 突触前神经元的 EV 前体和突触后分泌的 EV 中的 Syt4 均调节 NMJ。 突触生长和神经递质释放以活动依赖的方式，导致我的假设 CaMKII 调节 Syt4 EV 信号传导以促进结构和功能可塑性 在目标 1 中，我会问 CaMKII 是否如此。 控制我们之前确定的调节 EV 货物的内吞运输途径中的 EV 货物水平 在第二个目标中，我将阐明 CaMKII 和 Syt4 之间的关系并测试它们是否以某种方式起作用。 在目标 3 中，我将分离特定的 CaMKII。 这些目标将提供先进成像、 定量图像分析、果蝇遗传学和电生理学从中获得的信息。 该项目将为 CaMKII 如何调节 EV 贩运以促进突触可塑性提供新的见解，以及 确定神经系统中调节 EV 的新活动依赖性机制。