Zebrafish scn8aa sodium channels mediate the majority of sodium conductance, which is essential for the embryonic locomotor activities. Here, we investigated the transcriptional regulation of scn8aa in developing zebrafish embryos by constructing a GFP reporter driven by a 15-kb fragment of scn8aa gene designed as scn8aa:GFP. GFP expression patterns of scn8aa:GFP temporally and spatially recapitulated the expression of endogenous scn8aa mRNA during zebrafish embryonic development, with one exception in the inner nuclear layer of the retina. Three novel elements, along with an evolutionarily conserved element shared with mouse SCN8A, modulated neuronal-specific expression of scn8aa. The deletion of each positive element reduced the expression levels in neurons without inducing ectopic GFP expression in non-neuronal cells. Our results demonstrate that these four regulatory elements function cooperatively to enhance scn8aa expression in the zebrafish nervous system. Developmental Dynamics 237:2554–2565, 2008. © 2008 Wiley-Liss, Inc.