Spatial control of membrane traffic by septin GTPases

Septin GTPases 对膜运输的空间控制

• 批准号: 10389249
• 负责人: Elias T Spiliotis
• 金额: $ 15万
• 依托单位: DREXEL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10389249
• 关键词: 10 year old; 3-Dimensional; Administrative Supplement; Alzheimer' s Disease; Apical; Brain Injuries; Cells; Cues; Data; Defect; Destinations; Development; Disease; Docking; Epithelial; Family; Funding; GTP-Binding Proteins; Guanosine Triphosphate Phosphohydrolases; Homeostasis; Image; Knowledge; Laser Scanning Confocal Microscopy; Lateral; Lysosomes; Maintenance; Malignant Neoplasms; Membrane; Membrane Protein Traffic; Metabolic Diseases; Microscope; Microtubules; Morphogenesis; Nerve Degeneration; Neurons; Organ; Organoids; Outcome; Output; Parkinson Disease; Pathogenesis; Physiological; Positioning Attribute; Productivity; Protein Family; Protein Sortings; Proteins; Research; Resolution; Route; Signal Transduction; Specificity; System; Time; Transmembrane Transport; Vesicle; cell type; detector; human disease; imaging capabilities; improved; insight; instrumentation; neurotransmission; novel; polarized cell; programs; quantum; reconstruction; response; spatiotemporal; temporal measurement; trafficking

Project Summary/Abstract Spatial control of membrane traffic is essential for the morphogenesis and maintenance of polarized cell types such as epithelia and neurons, whose organization is integral to organ homeostasis and neurotransmission. Many advances have been made in understanding membrane traffic at points of origin (protein sorting, vesicle formation) and destination (vesicle docking/fusion). However, key challenges remain in understanding how long-range transport is spatially controlled en route to destination. The central hypothesis of our studies is that septins, a family of multimeric GTP-binding proteins, which associate with distinct subsets of microtubules and membrane domains, comprise a novel regulatory module for the spatial guidance of membrane traffic. Here, we request an administrative supplement for the acquisition of fast and super-resolution laser scanning confocal microscopy, which will improve the imaging capabilities, quality and data output (productivity) of our R35-funded projects. In investigating whether there is specificity between septins and distinct routes of polarized traffic (e.g., apical vs. basolateral), the new instrumentation will enable to image fixed and live epithelial sheets and organoids faster and with less photodamage as well as unprecedented resolution (100 nm lateral, 200 nm axial). In our studies of how lysosome trafficking and positioning is regulated by septins at steady state and in response to cellular cues, the new instrumentation will enable to track endolysosomes with unprecedented spatiotemporal resolution in 2D and 3D using detectors with superior quantum efficiency. Through the use of advanced modulations such as dynamic enhancement and adaptive image quality determination and reconstruction, we will be able to acquire high-quality time-lapse data, which currently is not possible with core confocal and spinning disk instrumentation that is over ten years old. Outcomes will shed valuable insights into the mechanisms that direct the transport of membrane vesicles and endolysosomes in response to morphogenetic and physiological signals. The proposed studies will also advance our understanding of septins as spatial regulators of intracellular organization, bearing significance on diseases triggered and/or exacerbated by abnormalities in septin expression.

项目总结/摘要 膜运输的空间控制对于形态发生和维持至关重要 极化细胞类型，如上皮细胞和神经元，其组织是器官的组成部分， 稳态和神经传递。在理解上已经取得了许多进展 膜运输在起点（蛋白质分选，囊泡形成）和终点（囊泡 对接/融合）。然而，在了解远距离迁移如何影响生物多样性方面仍然存在重大挑战。 在前往目的地的途中受到空间控制我们研究的中心假设是， 一个多聚体GTP结合蛋白家族，与不同的 微管和膜结构域，包括一个新的调控模块的空间 膜交通的引导。在此，我们要求对收购进行行政补充 快速和超分辨率激光扫描共聚焦显微镜，这将提高成像 我们的R35资助项目的能力、质量和数据输出（生产力）。在调查 在分隔符和极化业务的不同路由之间是否存在特异性（例如，心尖 vs.基底外侧），新仪器将能够对固定和活体上皮片进行成像， 类器官更快，光损伤更少，分辨率前所未有（100 nm 横向，200 nm轴向）。在我们的研究中，溶酶体的运输和定位是如何被 septins在稳定状态和响应细胞的线索，新的仪器将能够 在2D和3D中以前所未有的时空分辨率跟踪内溶酶体， 具有上级量子效率的探测器。通过使用先进的调制，如 动态增强和自适应图像质量的确定和重建，我们将 能够获取高质量的延时数据，这是目前核心共聚焦无法实现的 和旋转圆盘仪器已经有十多年的历史了结果将失去宝贵的 深入了解直接膜囊泡的运输机制， 内溶酶体对形态发生和生理信号的反应。拟议的研究 也将促进我们对septins作为细胞内组织的空间调节因子的理解， 对由septin异常触发和/或加重的疾病具有重要意义 表情