Decipher membrane patterns in situ with super-resolution and dynamic microscopy

使用超分辨率和动态显微镜原位解读膜图案

• 批准号: 8358427
• 负责人: Bjoern F Lillemeier
• 金额: $ 288万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-30 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8358427
• 关键词: Adverse effects; Architecture; Cell membrane; Color; Cytokine Signaling; Defect; Disease; EGF gene; Environment; Fluorescence; Foundations; Hand; Image; In Situ; Life; Membrane; Membrane Proteins; Methods; Microscopy; Molecular; Movement; Optics; Pattern; Plant Roots; Process; Research; Resolution; Signal Pathway; Signal Transduction; Specimen; Spectrum Analysis; Technology; Visual; abstracting; base; fluorophore; insight; new technology; novel; public health relevance; segregation; theories

DESCRIPTION (Provided by the applicant) Abstract: The inability of current super-resolution methods to generate multi color in situ images and dynamic information limits their impact. The challenge at hand is to advance this purely descriptive technology to study the mechanisms of highly dynamic and rare molecular processes. We will approach the task of visualizing two different colors in live specimens through the combination of different fluorophores and new acquisition strategies. Information on the interplay between molecule distributions and movement will be derived from simultaneous analyses of molecule dynamics by fluorescence cross correlation spectroscopy. With this novel technology, we will study the architecture of the plasma membrane and its effects on membrane associated signaling. This research will provide visual and mechanistic insights necessary to develop a unified plasma membrane theory. Specifically, we will explore the segregation of all membrane proteins into domains and the basis of their distinct confinement. In addition, the proposed research wil disect the spatio-temporal mechanisms of different signaling pathways (TCR, EGF and cytokine signaling). We expect to discover novel control principles in signaling that are rooted in the architecture of the plasma membrane. Thus, our studies have the potential to revolutionize our understanding of the molecular underpinnings regulating membrane- associated signaling. This is crucially important in view of the large number of diseases associated with membrane signaling defects. Our studies will verify the potential for the modulation of membrane associated signaling through changes in the plasma membrane organization. Public Health Relevance: Studying the plasma membrane structure and signaling mechanisms rooted in it lays the foundation for insights into a large number of diseases that are caused by defects in membrane signaling. The proposed research aims to establish technology that can determine the spatial and dynamic underpinnings of membraneassociated signaling based on super-resolution and cross-correlation optical microscopy. It will provide new avenues for modulating cellular signaling through changes in its environment, a more 'gentle' adjustment without causing the dramatic side effects of current approaches.

描述（申请人提供） 摘要：当前的超分辨率方法无法生成多色原位图像和动态信息限制其影响。眼前的挑战是推进这种纯粹的描述性技术，以研究高度动态和罕见的分子过程的机制。我们将通过不同的荧光团和新的采集策略的结合来实现在实时标本中可视化两种不同颜色的任务。关于分子分布与运动之间相互作用的信息将通过荧光互相关光谱法对分子动力学的同时分析得出。借助这项新技术，我们将研究质膜的结构及其对膜相关信号的影响。这项研究将提供开发统一的质膜理论所需的视觉和机械见解。具体而言，我们将探索所有膜蛋白对域的隔离及其独特限制的基础。此外，拟议的研究将消除不同信号通路（TCR，EGF和细胞因子信号传导）的时空机制。我们希望发现植根于质膜结构的信号传导中的新颖控制原理。因此，我们的研究有可能彻底改变我们对调节膜相关信号的分子基础的理解。鉴于与膜信号缺陷相关的大量疾病，这至关重要。我们的研究将通过质膜组织的变化来验证调节膜相关信号的潜力。 公共卫生相关性：研究其质膜结构和信号传导机制奠定了对大量疾病的见解，这些疾病是由膜信号传导中缺陷引起的。拟议的研究旨在建立可以根据超分辨率和互相关光学显微镜确定膜相关信号的空间和动态基础的技术。它将为通过环境变化调节细胞信号传导提供新的途径，而不必引起当前方法的显着副作用。