A novel localization based biosensor, SEDAR, visualizes spatio-temporal dynamics of activated Ras in endogenous Drosophila tissues

摘要: Chemokine signaling via growth factor receptor tyrosine kinases (RTKs) regulates development, differentiation, and disease implying that it is involved in a myriad of cellular processes. A single RTK, for example the Epidermal Growth Factor Receptor (EGFR), used repeatedly multitude developmental programs. Quantitative differences magnitude duration RTK can bring about different outcomes. Understanding this complex signals requires real time visualization signal. To visualize spatio-temporal dynamics, biosensor called SEnsitive Detection Activated Ras (SEDAR) was developed. It localization-based sensor binds to activated directly downstream endogenous RTKs. This binding reversible SEDAR expression did not cause any detectable perturbation Using SEDAR, guidance visualized during mediated chemotaxis border cells Drosophila ovary. localized both leading rear end cluster but polarized at edge cluster. Perturbation RTKs led delays forward migration correlated with loss polarization Gliding or tumbling behavior related high low respectively, cell showing signal plasma membrane provided information migratory behavior. Further, localization detected EGFR five distinct contexts. Hence novel could be as valuable tool study dynamics activation RTKs, three-dimensional tissues, an unprecedented spatial temporal resolution.