A contextual framework for characterizing motility and chemotaxis mutants in Dictyostelium discoideum.

In the natural aggregation process, Dictyostelium amoebae relay the cAMP signal outwardly through the cell population as symmetric, nondissipating waves. Each cell in turn responds in a specific manner to the different phases of the wave. In the front of each wave, each cell experiences an increasing temporal gradient and positive spatial gradient of cAMP; at the peak of each wave, each cell experiences a cAMP concentration inhibitory to locomotion; and in the back of each wave, each cell experiences a decreasing temporal and negative spatial gradient of cAMP. Protocols are described to analyze the basic motile behavior of mutant cells in the absence of a chemotactic signal, and to test the responsiveness of mutant cells to the individual temporal, spatial and concentration components of a natural wave. The results of such an analysis can then be used to develop realistic models of cell motility and chemotaxis. Examples are described in which this contextual framework has been applied to mutant cell lines. The results of these mutant studies result in a model in which independent parallel regulatory pathways emanating from different phases of the wave effect different phase-specific behaviors.