Drosophila tracheal cytonemes in long range FGF signaling

项目总结/摘要 这是一份K99/R 00奖的申请书，申请人是大学的博士后Sougata Roy博士。 来自加州的弗朗西斯科。罗伊博士在生物化学、分子生物学、发育 遗传学和细胞生物学，并寻求在他提出的目标先进的成像技术的额外培训。 罗伊博士与他的主要导师托马斯B博士组建了一个指导小组。Kornberg教授 心血管研究所，一位知名的发育生物学家，以其 在网状和Hedghog信号通路的基本贡献，并为发现 和两名顾问：大卫阿加德博士，生物化学和生物物理学系教授， 加州大学旧金山分校，世界著名的，在他的开拓性工作的一部分，与方法的发展，自动EM 断层扫描、3D重建和结构照明;以及加州大学旧金山分校教授罗恩瓦尔博士 细胞和分子药理学系，以其对理解 细胞骨架动力学和细胞内的空间组织和运动。 罗伊博士的长期目标是了解细胞的细胞和分子机制 远距离沟通。他的建议集中在细胞素的作用上，细胞素是一种 信号丝状伪足，并对FGF的机制，在后生动物的关键信号蛋白之一， 在果蝇的发育过程中，细胞穿过组织到达果蝇的肺或气囊原基。人们早就 接受信号蛋白如FGF在细胞外空间扩散以产生浓度梯度， 这些梯度调节响应细胞的生长和模式。虽然多年来丝状伪足 果蝇细胞丝的发现和特征表明， 一种用于信号蛋白移动的替代模型-信号蛋白可以以 在直接接触点的控制方式。控制信号水平和目标是至关重要的， 和疾病状态;一个说明性的例子是肺增生，其在小鼠中可以由以下任一种引起： FGF 10信号传导的降低或升高。本申请中提出的工作研究了 细胞丝接触，其是信号蛋白从源细胞转移到靶细胞的位点，以及 调节FGF运输和活性的机制。这些研究将采用果蝇遗传学， 重组工程，RNAi筛选和最先进的显微镜技术，包括荧光显微镜， 共聚焦显微镜、电子显微镜/三维电磁断层扫描、结构照明显微镜 (SIM)在UCSF。 这项研究将大大提高对这一机制和作用的认识， 细胞丝介导的信号在肺的发展，并预计将形成的基础上，R 01赠款 在裁决结束前提出申请。

PROJECT SUMMARY/ABSTRACT This is an application for a K99/R00 award for Dr. Sougata Roy, a post-doctoral fellow at the University of California, San Francisco. Dr. Roy is experienced in biochemistry, molecular biology, developmental genetics and cell biology and seeks additional training in advanced imaging techniques in his proposed goals. Dr. Roy has assembled a mentoring team with his primary mentor, Dr. Thomas B. Kornberg, Professor in the Cardiovascular Research Institute, a well-recognized developmental biologist and known for his fundamental contributions in the engrailed and Hedghog signaling pathways and for the discovery of cytonemes; and two advisors: Dr. David Agard, Professor in the Department of Biochemistry and Biophysics, UCSF, world-renowned, in part of his pioneering work with the development of methods for automated EM tomography, 3D reconstruction and structured illumination; and Dr. Ron Vale, Professor in the UCSF Department of Cellular and Molecular Pharmacology, world renowned for his contributions to understanding cytoskeletal dynamics and spatial organization and movement within cells. Dr. Roy's long-term goal is to understand the cellular and molecular mechanisms which cells use to communicate over long distances. His proposal focuses on the roles of cytonemes, which are a type of signaling filopodia, and on the mechanisms by which FGF, one of the key signaling proteins in metazoan development, moves across tissues and reaches Drosophila lung or air-sac primordium. It has long been accepted that signaling proteins like FGF diffuse in extracellular space to generate concentration gradients and that these gradients regulate growth and patterning of responding cells. Although for many years filopodia have been implicated principally in migration, the discovery and characterization of Drosophila cytonemes suggested an alternative model for movement of signaling proteins - that signaling proteins may be delivered in a controlled fashion at points of direct contact. Controlling signaling levels and targets is critical to both normal and disease states; an illustrative example is pulmonary hyperplasia, which in the mouse can result from either reduction or elevation of FGF10 signaling. The work proposed in this application investigates the nature of the cytoneme contacts that are the sites of transfer of signaling proteins from source to target cells, as well as the mechanisms that regulate FGF transport and activity. The studies will employ Drosophila genetics, recombineering, RNAi screen and state-of-the-art microscopy techniques, including fluorescent microscopy, confocal microscopy, Electron Microscopy/3-Dimension EM-tomography, structured illumination microscopy (SIM) available in UCSF. The proposed research will significantly improve understanding of the mechanism and role of cytoneme-mediated signaling during lung development and is expected to form the basis for an R01 grant application before the end of the award.