Drosophila tracheal cytonemes in long range FGF signaling

描述（由申请人提供）：这是一个K99/R 00奖博士Sougata罗伊，博士后研究员在加州大学，旧金山弗朗西斯科的应用程序。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方面经验丰富，并在他提出的目标中寻求先进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，心血管研究所教授，一位公认的发育生物学家，以其在交织和Hedghog信号通路以及细胞素发现方面的重要贡献而闻名;以及两位顾问：大卫阿加德博士，教授在生物化学和生物物理学系，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，与方法的自动电磁断层扫描，3D重建和结构化照明;以及UCSF细胞和分子药理学系教授罗恩瓦尔博士，他因理解细胞骨架动力学和空间组织以及细胞内运动而闻名。 罗伊博士的长期目标是了解细胞用于长距离通信的细胞和分子机制。他的建议集中在细胞丝的作用，这是一种信号丝状伪足，并在FGF的机制，在后生动物的发展中的关键信号蛋白之一，跨组织移动，并达到果蝇肺或气囊原基。长期以来，人们一直认为信号蛋白如FGF在细胞外空间扩散以产生浓度梯度，并且这些梯度调节响应细胞的生长和图案化。虽然多年来丝状伪足主要与迁移有关，但果蝇细胞丝的发现和表征表明了信号蛋白运动的另一种模式-信号蛋白可以在直接接触点以受控方式传递。控制信号传导水平和靶标对于正常和疾病状态都是至关重要的;说明性实例是肺增生，其在小鼠中可由FGF 10信号传导的降低或升高引起。本申请中提出的工作研究了细胞丝接触的性质，细胞丝接触是信号蛋白从源细胞转移到靶细胞的位点，以及调节FGF转运和活性的机制。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EM断层扫描，结构照明显微镜（SIM）在UCSF可用。 拟议的研究将显着提高对肺发育过程中细胞丝介导的信号传导机制和作用的理解，预计将在该奖项结束前形成R 01资助申请的基础。

DESCRIPTION (provided by applicant): 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.