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.

项目摘要 细胞外蔬菜（EV）是膜结合的小室，可在细胞之间交换材料， 并在神经系统中的细胞通信中发挥重要作用。神经元EV介导活性 - 依赖性突触可塑性，以及与 神经退行性疾病，例如阿尔茨海默氏病和帕金森氏病。 EV生物发生和 分泌是通过内体运输发生的，但是神经元活性调节这些细胞的机制 生物过程尚未得到很好的特征。通过识别与活动有关的机制影响者 神经元EV动力学，我们将对神经系统中的基本和病理过程有更深入的了解。 大多数EV研究都利用体外非神经细胞模型分离出异质的EV种群， 直接测量神经元EV生物发生和运输机制挑战。果蝇 神经肌肉连接（NMJ）是一种强大的体内模型，非常适合操纵和 可视化电动汽车货物。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），一种参与突触可塑性结构和功能形式的关键突触蛋白 在电动汽车贩运中的作用。具体而言，CAMKII无效突变体显示出显着提高的EV货物的水平 在突触前神经元和伴有后分泌的电动汽车中的两个EV前体中的SYT4。 SYT4调节NMJ 突触生长和神经递质以活性依赖的方式释放，导致我的假设是 CAMKII调节SYT4 EV信号，以促进结构和功能可塑性。在AIM 1中，我会问Camkii是否 在我们以前确定的内吞贩运途径中控制EV货物水平 贩运。在第二个目标中，我将阐明Camkii和Syt4之间的关系，并测试它们是否在 共享促进结构和功能可塑性的途径。在AIM 3中，我将隔离CAMKII 涉及EV货物和贩运调节的功能。这些目标将提供高级成像的培训， 定量图像分析，果蝇遗传学和电生理学。从此获得的信息 项目将提供有关CAMKII如何调节电动汽车贩运以促进合成可塑性的新见解，并且 确定调节神经系统中电动汽车的新活动依赖性机制。