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 Merit This 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 Impact This 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.
