During gastrulation, germ layers are formed as prospective mesodermal and endodermal cells internalize and come to underlie the ectoderm [1-9]. Despite the pivotal role of gastrulation in animal development, the cellular interactions underlying this process are poorly understood. In zebrafish, mesoderm and endoderm formation requires the Nodal signals Cyclops and Squint and their cofactor One-eyed pinhead (Oep) [10-14]. We found that marginal cells in maternal-zygotic oep (MZoep) mutants do not internalize during gastrulation and acquire neural and tail fates at the expense of head and trunk mesendoderm. The lack of internalization in MZoep embryos and the cell-autonomous requirement for oep in Nodal signaling enabled us to test whether internalization can be achieved by individual cells or whether it depends on interactions within a group of cells. We found that individual MZoep mutant cells transplanted to the margin of wild-type blastula embryos initially internalize with their neighbors but are unable to contribute to the mesendoderm. In the reciprocal experiment, single wild-type cells transplanted to the margin of MZoep mutant embryos autonomously internalize and can express the mesendodermal markers axial/foxA2 and sox17. These results suggest that internalization and mesendoderm formation in zebrafish can be attained autonomously by single cells.