Quantitative analysis of bicoid degradation and diffusion kinetics in a developing embryo.

DURRIEU, LUCÍA; SAUNDERS, TIMOTHY; KIRRMAIER, DANIEL; KNOP, MICHAEL; HUFNAGEL, LARS

Mont Ste. Odile

Encuentro; Cell Biology and Biophysics meeting; 2014

EMBL

Presentación oral--In Drosophila development, the gradient of the morphogen Bicoid determines the patterning of the embryo across the anterior-posterior axis. Despite the fact that it has been extensively studied, the molecular mechanisms that control the shaping of this gradient are still not understood. For two important factors, the diffusion and degradation, the reaction rates are still controversial and very little is known about the machinery involved. We are currently studying both processes through various biochemical and imaging approaches. Here we present the results of fusing Bicoid to a fluorescent timer -two single color fluorescent proteins with different maturation kinetics-. We show that using this timer coupled with a spatial model allows us to determine both Bicoid´s degradation and diffusion rates from single snapshot images. We combine the fluorescent timers with light-sheet microscopy to get images of the whole embryo with subcellular resolution, high signal to noise ratio, and minimal phototoxicity. This tool will give us a complete map of Bicoid kinetics in space and time, and provide a rich read-out for future studies of its underlying molecular mechanisms.