Dokument

Summary:
This work provides new insights into the dynamics and structure of cellular aggregation. Starting from cell motility which is necessary to get the cells into close proximity it presents new tools for visualization, analysis and modeling of aggregation processes. While a lot of work has been done in the field of microbial and amoeboid motility, there is a lack in theoretical understanding of mammalian cell motion, especially concerning directed migration stirred by external cues. To close this gap I developed a two-dimensional generic model based on mechanical cell-substrate interactions. This model facilitates the discrete nature of the motion cycle of mammalian cells by a randomized growth of protrusions and their retraction depending on the strength of an external cue. This model is capable of reproducing most experimental observations, especially the behavior at sharp changes in strength of the external cues, and provides an explanation for the attachment of the lagging cell pole as it increases the efficiency of gradient sensing. Furthermore, I introduce new experimental methods to visualize and analytical toolkits to analyze the structure of the highly irregular cell aggregates. These approaches were tested in two example cases: the two dimensional aggregation of mouse embryonic fibroblast (MEF)cells and the flocculation of S. cerevisiae mediated by the sugar-dependent adhesion protein Flo5. While it was possible to achieve temporal information of the MEF cell aggregation, the flocculation of S. cerevisiae is not accessible in this way. The time-lapse microscopy series indicate a subdivision of MEF cell aggregation into a spreading and a contraction phase. In addition, the data shows that there is a dependency of the aggregate’s structure on its size with a sharp transition from a linear dependency to a constant structure. The three-dimensional imaging of immobilized flocs using a confocal laser scanning microscope provided information about the structural properties of yeast flocs. The most important findings are that the flocs are self similar fractal structures and that cheater cells, i.e. cells that do not produce the necessary binding proteins but benefit from the altruistic behavior of producing cells, are largely underprivileged in the process. This indicates that, even though flo5 does not qualify as a “green beard gene” by definition, the benefits of the resulting altruistic behavior are strongly shifted in favor of the producing cells by the aggregation mechanism.

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