Lateral propagation of EGF signaling after local stimulation is dependent on receptor density

We analyzed lateral propagation of epidermal growth factor (EGF) signaling in single live COS cells following local stimulation, achieved by the use of laminar flows containing rhodamine-labeled EGF. The spatiotemporal pattern of EGF signaling was visualized by fluorescent indicators for Ras activation and tyrosine phosphorylation. Contrary to the findings in previous reports, both signals were localized to the stimulated regions in control COS cells expressing EGF receptor at the basal level. However, the signals spread over the entire cell when EGF receptors were overexpressed or when receptor/ligand endocytosis was blocked. We thus present evidence that ligand-independent propagation of EGF signaling occurs only when the receptor density on the plasma membrane is high, such as in carcinoma cells.