Microfluidic Temperature Steps to Understand Robustness of Embryonic Development

项目总结。这项提案描述了一个多学科的研究计划，旨在开发， 验证和传播微流控技术，使果蝇活胚胎的发育成为 使用温度步长在空间和时间上进行控制。果蝇胚胎发育的过程是 坚固耐用-即使在温度等不同的环境条件下，它也能准确地工作。而当 果蝇发育过程中存在的许多单个分子的功能是已知的，还不清楚这些分子是如何 分子共同努力，使发育网络变得强大。新的微流控技术可以 通过差异化地控制不同部位的温度，活着的胚胎可能会成为 确定对发展的这种稳健性负责的机制。具体目标1侧重于 开发新的微流控技术，利用层流产生陡峭的温度阶跃 在胚胎周围，胚胎的一部分将在一个层流的温度下发育 胚胎的另一部分将在第二层流的较低温度下发育。 溪流。胚胎表面的温度分布将用 数值模拟和共聚焦显微镜。实时成像胚胎暴露在 温度步骤在识别动态过程中至关重要，例如蛋白质浓度的变化 时间的作用。特定目标2采用在特定目标1中开发的技术来挖掘和双光子 显微镜下为了实时成像胚胎发育的温度步长。建议 对特定目标3的研究将验证在特定目标1和特定目标1中开发的微流控技术 目的2通过回答关于稳健性机制的四个重要问题，这两个问题都是在分子 在核一级和核区一级。 相关性：了解发展对于了解人类疾病和状况至关重要 由发育和分化途径中的缺陷和错误引起的(如癌症和衰老)。 在模式生物中的研究进展？特别是果蝇黑腹果蝇导致了 更好地了解人类发展，因为发展的许多基本机制都是 在有机体中类似。本提案中开发的微流控技术将使人们能够理解 果蝇发育网络无差错运行的机制 这项技术将可扩展到测试和理解开发过程中的错误 其他模式生物

Project summary. This proposal describes a multi-disciplinary research program that aims to develop, validate and disseminate microfluidic technology to allow development of a live Drosophila embryo to be controlled in space and time using temperature steps. The process of Drosophila embryo development is robust - it works precisely even under varying environmental conditions such as temperature. While the function of many individual molecules present in Drosophila development is known, it is unknown how these molecules work together to make developmental network robust. New microfluidic technology that can differentially control temperature around different parts a living embryo could become a powerful tool in determining the mechanisms responsible for this robustness of development. Specific Aim 1focuses on development of new microfluidic technology that will use laminar flow to create a sharp temperature step around the embryo, where one part of the embryo will develop at the warmer temperature of one laminar stream, and the other part of the embryo will develop at the cooler temperature of the second laminar stream. The temperature profile at the surface of the embryo will be quantitatively characterized using numerical simulations and confocal microscopy. Real-time imaging of an embryo being exposed to temperature step is critical in identifying dynamic processes such as changes in protein concentration as a function of time. Specific Aim 2 adapts technology developed in Specific Aim 1to DIG and 2-photon microscopy in order to image embryonic development in the temperature step in real time. Proposed research in Specific Aim 3 will validate the microfluidic technology developed in Specific Aim 1and Specific Aim 2 by answering four important questions concerning the mechanism of robustness, both at the molecular level and at the level of nuclear divisions. Relevance: Understanding development is essential for understanding of human diseases and conditions caused by defects and errors in developmental and differentiation pathways (such as cancer and aging). Studying development in model organisms¿in particular, the fruit fly Drosophila melanogaster¿leads to a better understanding of human development, since many parts of the basic machinery of development are similar among organisms. The microfluidic technology developed in this proposal will enable understanding of the mechanism that provides error-free operation of developmental network in the fruit fly Drosophila melanogaster, and this technology will be extendable to testing and understanding errors in development of other model organisms