Formation of mesoderm in Xenopus embryos is the result of an induction event in which peptides such as FGF or activins have been implicated. It was recently demonstrated, by the ectopic expression of a truncated activin receptor, that activin receptor signaling pathways are involved in the processes of mesoderm and axis formation in vivo. However, this approach does not directly address the role of activin itself nor the involvement of activins in the formation of mesoderm in embryos from other vertebrates. In addition, activins are expressed maternally as a protein component of the egg as well as transcribed zygotically, and it is not clear which of the two forms is involved in mesoderm formation. To address those three issues, we analyzed the role of activins in the development of fish embryos by generating two activin dominant-negative variants. One of the variants behaves as an inhibitor of activin protein. The second variant was found to deplete the activin pool when cotranslated with wild-type activin. Injection of RNA encoding these variants into the two-cell embryo of the small teleost fish Oryzias latipes (Japanese medaka) demonstrates that only the maternally provided activin protein is required for mesoderm and axis formation in fish in vivo.