EFA6A is a guanine nucleotide exchange factor (GEF) that can activate ADP-ribosylation factor 6 (ARF6) in vitro, with prominent expression in the forebrain including the hippocampal formation. In this section, we describe the neuronal transfection method and show that the overexpression of a catalytically inactive mutant of EFA6A induces a prominent dendritic formation of the primary hippocampal neurons, suggesting the intimate involvement of EFA6A in the regulation of neuronal dendritic development. This reliable and consistent neuronal transfection method will also be applicable for the vector-based RNA interference method.