The Wnts are a family of secreted glycoproteins involved in cell-cell signalling and pattern formation during development, although the extent to which various Wnts are functionally equivalent remains unclear. We have cloned zebrafish (Danio rerio) wnt4, characterized its expression, and compared its activity relative to other Wnts. The wnt4 transcript is first detected early in somitogenesis, in the dorsocaudal region of the forebrain, and then appears in the dorsal and lateral regions of the caudal hindbrain and neural keel. During somitogenesis, wnt4 appears in the floor plate, and this expression is absent in cyclops mutants, which lack floor plate. wnt4 is also expressed in the developing pronephros and gill slit. To characterize the biological activity of wnt4, synthetic zebrafish wnt4 mRNA was injected into embryos of zebrafish and Xenopus laevis. The phenotypic effects of misexpression in the zebrafish include cyclopia, misfolding in the brain, and an anteriorly forking notochord. Comparison of the phenotypes arising from misexpression of wnt4 and Xwnt-5A in both organisms suggests close parallels in the response to these Wnts. Our data suggest that wnt4, like Xwnt- 5A, inhibits cell movements, and that these Wnts define a functional class distinct from the class which includes Wnt-1, Xwnt-3A and Xwnt-8.