CAREER: Engineering principles within cell-cell communication networks in animal development

1254344 Reeves细胞间信号的调节在许多生物学领域都是至关重要的，包括发育、干细胞生物学和成体组织的适当维护。 在发育过程中，组织通过长距离但高度可变的信号形成模式，这些信号指导不同位置的基因表达差异。 在首席研究员的实验室中，首要目标是将工程原理与调控机制联系起来，以确保在面对高度可变的细胞间信号时可靠的基因表达模式。 拟议的工作旨在研究生物调节机制中工程原理的两个具体实例：反馈控制和噪声过滤。果蝇果蝇被用作模式生物，因为它在所需的复杂性（作为多细胞生物）和易于操作之间取得了平衡，具有先进的遗传和转基因工具包。 在早期（1-3小时）果蝇胚胎中，背腹轴由转录因子Dorsal形成。 在以前的工作中，发现Dorsal建立了强大的基因表达模式，即使在面对高内在噪声和外在变异。负反馈相互作用与二级信号传导途径，骨形态发生蛋白（BMP）的途径，被假设为增加基因表达模式的鲁棒性，在背信号的变化。 此外，背侧靶基因在远远超出信号分子的空间梯度预期可靠作用的点的位置表达。 因此，噪音过滤能力的基因表达的尾部的背信号，噪音是最高的，将进行调查。 负反馈相互作用和噪声过滤将使用遗传和转基因操作，定量（实时）成像/基因表达输出模型，定量实时PCR和定量Western Blot进行研究。与拟议的实验研究相关，推广目标是通过突出生物系统中的工程原理的教育材料促进工程和生物学之间的协同作用。 将与负责监督K-12外联活动的NCSU组织“科学之家”合作，为高中生开发和传播一个充实模块。目标是培养未来的生物学专业，在高中初中和高中水平，是舒适的语言和工程和物理的观点。 PI教授的课程的本科生将作为助教参加。 在研究生阶段，将在NCSU远程教育计划的帮助下广泛传播一门侧重于组织图案化工程原理的专题课程。 这些教育材料，连同研究工作，将提供的角度和研究，了解，然后利用？自然工程？存在于生物系统中。

1254344ReevesRegulation of signaling between cells is crucial in many areas of biology, including development, stem cell biology, and proper maintenance of adult tissues. In development, tissues are patterned by long range, yet highly variable signals that direct differences in gene expression at different locations. In the Principal Investigator's lab, the overarching goal is to connect engineering principles to the regulatory mechanisms that ensure reliable gene expression patterns in the face of highly variable intercellular signals. The proposed work aims to study two specific instances of engineering principles in biological regulatory mechanisms: feedback control and noise-filtering. The fruit fly Drosophila melanogaster is used as a model organism, as it strikes a balance between the complexity needed (as a multicellular organism) and ease of manipulation, with advanced genetic and transgenesis toolkits. In the early (1-3 hours old) Drosophila embryo, the dorsal-ventral axis is patterned by the transcription factor Dorsal. In previous work, it was found that Dorsal establishes robust gene expression patterns even in the face of high intrinsic noise and extrinsic variability. Negative feedback interactions with a secondary signaling pathway, the Bone Morphogenetic Protein (BMP) pathway, are hypothesized to increase the robustness of gene expression patterns with respect to variations in Dorsal signaling. Furthermore, dorsal target genes are expressed in a location far beyond the point in which a spatial gradient of a signaling molecule is expected to reliably act. Thus, the noise-filtering capabilities of gene expression in the tail of Dorsal signaling, where noise is the highest, will be investigated. Both negative feedback interactions and noise-filtering will be studied using genetic and transgenic manipulations, quantitative (live) imaging/models of gene expression outputs, quantitative real-time PCR and quantitative Western Blot.Related to the proposed experimental studies, the outreach goal is to facilitate the synergy between engineering and biology through educational materials that highlight the engineering principles found in biological systems. An enrichment module for high school students will be developed and disseminated in partnership with the Science House, an NCSU organization that oversees K-12 outreach. The goal is to train future biology majors, at the high school junior and senior level, to be comfortable with the language and viewpoint of engineering and physics. Undergraduate students in courses taught by the PI will participate as teaching assistants. At the graduate level, a special topics course that focuses on the engineering principles in tissue patterning will be broadly disseminated with the help of the NCSU Distance Education Program. These educational materials, together with the research work, will provide the perspective and research to understand and then utilize the ?natural engineering? present in biological systems.