Digital image processing of optical density wave propagation in Dictyostelium discoideum and analysis of the effects of caffeine and ammonia

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Abstract
Waves of chemotactic movement during the early phase of aggregation in Dictyostelium discoideum were analyzed by digital image processing in a manner that immediately shows the following parameters: wave propagation velocity, period length, wave amplitude und wave shape. We have characterized the aggregation of AX-2 and the streamer F mutant NP 377 in terms of these parameters and investigated the influence of caffeine and ammonia. It was found that during normal aggregation oscillation frequency increases while at the same time wave propagation velocity decreases. Caffeine, a known inhibitor of cyclic AMP relay, reduces oscillation frequency and wave propagation velocity in a dose-dependent manner but most notably leads to the appearance of bimodal (harmonic) oscillations. These bimodal waves are also found in streamer F mutants without caffeine during early aggregation. The effect of caffeine is interpreted as an increase in the average chemotactic deadaptation time due to elevated cyclic GMP levels after a cyclic AMP stimulus. This increased deadaptation time results in some cells responding to every chemotactic signal, while others respond only to every second signal, leading to mixed population behavior and hence biphasic optical density waves. Ammonia has no significant influence on oscillation frequency and wave propagation velocity but shows a clear increase in the amplitude of the optical density waves. This may indicate a more vigorous chemotactic response by individual cells or a better synchronization of the responding cell populations due to shortened chemotactic deadaptation times.