Spatial control of membrane traffic by septin GTPases

项目摘要/摘要 膜交通的空间控制对于膜的形态发生和维持至关重要 指极化的细胞类型，如上皮细胞和神经元，其组织是器官不可或缺的 动态平衡和神经传递。在理解方面已经取得了许多进展 起始点(蛋白质分离、囊泡形成)和目的地(囊泡)的膜运输 对接/聚变)。然而，关键的挑战仍然是理解长距离运输是 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，毛细管膜蛋白， 一个多聚体GTP结合蛋白家族，与不同的GTP亚群相关 微管和膜域，组成了一个新的空间调控模块 引导膜上交通。在此，我们请求为此次收购提供行政补充 快速和超分辨率激光扫描共聚焦显微镜，这将改善成像 我们R35资助项目的能力、质量和数据输出(生产率)。在调查中 间隔素和不同的两极化流量路线之间是否有特异性(例如，顶端 与基底侧相比)，新的器械将能够成像固定的和活的上皮片和 有机化合物的速度更快，光损害更小，分辨率前所未有(100 nm 横向，200 nm轴向)。在我们关于溶酶体运输和定位如何受到 在稳定状态和对细胞提示的反应中，新的仪器将使 在2D和3D中以前所未有的时空分辨率追踪内溶体 具有卓越量子效率的探测器。通过使用高级调制，例如 动态增强和自适应图像质量的确定和重建，我们将 能够获得高质量的延时数据，这是目前核心共焦无法实现的 以及有十多年历史的旋转圆盘仪器。结果将产生宝贵的影响 对引导膜小泡运输的机制和 内溶酶体对形态发生和生理信号的反应。建议进行的研究 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， 间隔蛋白异常引发和/或加重的疾病的意义 表情。

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.