Multiplex analysis of boundary formation in the Drosophila embryo

该奖项是根据2009年美国复苏和再投资法案（公法111-5）资助的。在这项提议中，研究人员建议自动获取果蝇胚胎中单个细胞核的基因表达数据，并使用这种新的定量工具来分析两个细胞域之间尖锐直边界的形成。在这些研究中，他们的目标是创建计算工具，允许从果蝇胚胎的单个细胞中自动获取和分析多个荧光信号。这些进展应该允许使用基于多探针的原位杂交方法在单核分辨率下定量分析10-20个基因表达模式。提出的自动化多重方法将大大有助于创建全面的基因表达图谱，如艾伦研究所创建的大脑图谱，这对于理解基本的生物过程，如组织和器官的组织以及在发育模式进化过程中这些结构的变化非常重要。这些方法还可以确定在相似区域表达的不同基因的确切相对表达模式，这在目前是不可能的。因此，这种精细的基因表达图谱将使在胚胎内的单个细胞类型中同时重建多个基因的转录谱成为可能。软件和协议将提供给科学界，该项目将包括一名研究生和几名本科生。因此，它也将对社会有重要的教育效益。

This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).In this proposal, the investigators propose to automate the acquisition of gene expression data from individual cell nuclei in Drosophila embryos and use this new quantitative tool to analyze the formation of a sharp straight border between two domains of cells. Their goal in these studies is to create computational tools that will permit the automated acquisition and analysis of multiple fluorescent signals from single cells in the Drosophila embryo. These advances should allow for the quantitative analysis of 10-20 gene expression patterns at single nucleus resolution using a multi-probe based approach to in situ hybridization. The proposed automated multiplex methods should contribute substantially to enabling the creation of comprehensive gene expression atlases such as the Brain Atlases created by the Allen Institute, which are important for understanding basic biological processes such as organization of tissues and organs and changes in these structures during the evolution of developmental patterns. These methods should also make it possible to determine the exact relative expression patterns of different genes expressed in similar regions, which currently is not possible. Such refined gene expression maps will therefore make it possible to reconstruct transcription profiles for multiple genes simultaneously in individual cell types within an embryo. The software and protocols will be made available to the scientific community, and this project will include a graduate student and several undergraduates. Thus, it will also have important educational benefits for society.