Wnt signaling regulates embryonic patterning and controls stem cell homeostasis, while aberrant Wnt activity is associated with disease. One Wnt family member, Wnt3, is required in mouse for specification of mesoderm, and later regulates neural patterning, apical ectodermal ridge formation, and hair growth. We have identified and performed preliminary characterization of the zebrafish wnt3 gene. wnt3 is expressed in the developing tailbud and neural tissue including the zona limitans intrathalamica (ZLI), optic tectum, midbrain-hindbrain boundary, and dorsal hindbrain and spinal cord. Expression in these regions suggests that Wnt3 participates in processes such as forebrain compartmentalization and regulation of tectal wiring topography by retinal ganglia axons. Surprisingly, wnt3 expression is not detectable during mesoderm specification, making it unlikely that Wnt3 regulates this process in zebrafish. This lack of early expression should make it possible to study later Wnt3-regulated patterning events, such as neural patterning, by knockdown studies in zebrafish. Developmental Dynamics 238:1768–1795, 2009. © 2009 Wiley-Liss, Inc.