Analysis of PDZ-RGS3 Function in Ephrin-B Reverse Signaling

Transmembrane B ephrins and their Eph receptors signal bidirectionally. However, the molecular and cellular mechanisms of the “reverse signaling” by B ephrins are not fully understood. The identification of several B ephrin cytoplasmic domain-interacting proteins has begun to shed light on the signal transduction mechanisms and the cellular effects of B ephrin reverse signaling. This article describes the use of a Transwell migration assay system to characterize the function of a PDZ domain and a regulator of G-protein signaling (RGS) domain-containing protein, PDZ-RGS3, in mediating B ephrin reverse signaling in cultured neurons. The Transwell system discussed here can provide an effective cell-based assay for elucidating RGS protein-mediated signaling interactions in neurons.