The impact of spatial and temporal patterns on multi-cellular behavior.

摘要

What makes a fruit fly a fruit fly? Essentially this question stems from one of the most fascinating problems in biology: how a single cell (fertilized egg) can give rise to a fully grown animal. To be able to answer this question, the importance to how spatial and temporal patterns of gene and protein expression influence the development of an organism must be understood. After all, fruit fly larvae are segmented, while fertilized eggs are not. Pattern formation is fundamental to establishing this organization of the developing embryo with the ultimate goal being the precise arrangements of specialized cells and tissues within each organ in an adult organism.; The research presented here showcases the examples of studies that assess the impact spatial and temporal protein patterns have on the behavior of a collection of cells. By introducing new experimental, non-traditional techniques we developed model systems that allowed us to examine the dependence of the strength of adhesion of cells on the protein organization on sub-cellular, micron length scales, and to investigate how epithelial cell sheets coordinate their migration incorporating individual cell locomotion, molecular signal propagation and different boundary conditions.; The first part of this dissertation presents a photolithography-based silanization patterning technique that allowed us to homogeneously pattern large areas with high precision. This method is then applied to organizing cell adhesion-promoting proteins on surfaces for the purposes of studying and manipulating cell behavior. We show how the strength of adhesion is dependent on high local density of an adhesive extracellular matrix protein fibronectin. The varied appeal of this technique is exhibited by showing its applicability to pattern stretched DNA, too. The second part of this dissertation focuses on the impact of spatial and temporal propagation of a molecular signal (ERK 1/2 MAPK) in migrating epithelial sheets during wound healing. By tracking the motion of individual cells within the sheet under the three constructed conditions, we show how the dynamics of the individual cells' motion is responsible for the coordinated migration of the sheet in accordance with the activation of ERK 1/2 MAPK.