Hacking Morphogenesis: integrated, complex microsystems (ICS) for controlling embryonic pattern formation

该方案的重点是1)设计、制造和测试分布式微尺度致动器、细胞内荧光传感器分子和反馈控制的集成系统，该系统能够与斑马鱼胚胎组织进行化学接口；2)直接控制斑马鱼的中胚层构型和胚胎发育。所提出的集成微系统将具有一系列化学致动器，能够控制蛋白质向组织中的扩散。该系统将通过将荧光蛋白序列拼接到两个报告细胞发育状态的基因(GSC-GFP和NTL-RFP)中来检测发育组织中的蛋白质生产水平。该系统将持续监测转录因子的状态，并调整蛋白质的分布剂量。PI将解决发育生物学中一个基本的悬而未决的问题：图灵/迈哈特反应扩散模型是否适合描述斑马鱼的中胚层模式？我们将探索斑马鱼胚胎如何产生导致成熟生物体的复杂分化模式。智力价值这项研究将影响发育的基本生物学，并为与发育中的生物体的接口提供根本上的新技术。虽然许多成熟的方法允许研究人员可视化正在发育的组织，但在正在发育的组织中控制扩散环境的技术还处于初级阶段。集成传感器/执行器组件阵列将使我们能够以亚细胞分辨率探测细胞响应。广泛影响这项提议将促进不同实验室之间的交叉受精。这次交流将包括参与该项目的学生，并将包括午餐和与每个机构的核心研究人员社区的讲座。密歇根纳米制造设施(MNF)将接待范德比尔特公司的研究人员，并向他们介绍先进的微/纳米制造方法。在范德比尔特大学，博兹科教授将招待微系统工程专业的学生，在斑马鱼胚胎的生长、培养和观察方面提供实验室轮换和实验经验。

This proposal focuses on 1) the design, fabrication and testing of an integrated system of distributed microscale actuators, intracellular fluorescent sensor molecules and feedback control capable of chemically interfacing with developing Zebrafish embryonic tissue, 2) direct control of mesodermal patterning and embryonic development in Zebrafish.The proposed integrated microsystem will have an array of chemical actuators capable of controlling the diffusion of proteins into tissue. The system will sense the levels of protein production in the developing tissue by splicing fluorescent protein sequences into two genes that report on the developmental state of the cell (gsc-GFP and ntl-RFP). The system will continuously monitor the state of the transcription factors and adjust distributed dosing of proteins. The PIs will address a fundamental open question in developmental biology: is a Turing / Meinhardt reaction-diffusion model appropriate to describe mesodermal patterning in Zebrafish? We will explore how the Zebrafish embryo generates the complex patterns of differentiation that lead to a mature organism. Intellectual MeritThis research will impact the basic biology of development and provide fundamentally new technologies for interfacing with developing organisms. While a number of mature methods allow the researcher to visualize developing tissue, technologies for controlling the diffusible environment in developing tissue are in their infancy. Integrating arrays of sensor/actuator components will allow us to probe cell responses with sub-cellular resolution. Broad ImpactThis proposal will foster cross-fertilization between the different laboratories. This exchange will include the students working on the project and will include brown bag lunches and talks to the core community of researchers at each institution. The Michigan Nanofabrication Facility (MNF) will host Vanderbilt researchers and introduce them to advanced micro/nano-fabrication methods. At Vanderbilt University, Prof. Boczko will host microsystems engineering students, providing lab rotations and experimental experience in the growth, culture and observation of Zebrafish embryos.