The detection of gradients of chemotactic cues is a common task for migrating cells and outgrowing axons. Eukaryotic gradient detection employs a spatial mechanism, meaning that the external gradient has to be translated into an intracellular signaling gradient, which affects cell polarization and directional movement. The sensitivity of gradient detection is governed by signal amplification and adaptation mechanisms. Comparison of the major signal transduction pathways underlying gradient detection in three exemplary chemotaxing cell types, Dictyostelium, neutrophils, and fibroblasts and in neuronal growth cones, reveals conserved mechanisms such as localized PI3 kinase/PIP3 signaling and a common output, the regulation of the cytoskeleton by Rho GTPases. Local protein translation plays a role in directional movement of both fibroblasts and neuronal growth cones. Ca(2+) signaling is prominently involved in growth cone gradient detection. The diversity of signaling between different cell types and its functional implications make sense in the biological context.