Amacrine cells are located almost exclusively in the inner nuclear layer (INL) of the retina, but they express a variety of neurotransmitters. To begin to elucidate the relative roles of the local environment and cell lineage in determining the different neurotransmitter subtypes of amacrine cells, we combined lineage tracing and immunocytochemical techniques to map the spatial distribution and clonal origin of dopamine (DA) and neuropeptide Y (NPY) amacrine cells in Xenopus tadpole retina. At the earliest period of neurotransmitter expression, both DA and NPY amacrine cells were distributed preferentially in center and intermediate annular regions, and in anterior and dorsal quadrants. Most of the DA and NPY cells first emerged as scattered cells and later as clusters (of 2 or more cells) that increased in number and size up to premetamorphic stages. These results suggest that DA and NPY amacrine subtypes may be influenced by environmental cues localized to specific regions of the retina. Lineage analysis showed that the percentage of DA or NPY amacrine cells produced by most blastomere progenitors is significantly different from that predicted by the number of cells in the retina produced by those blastomeres. Only two blastomeres produced over 90% of the DA amacrine cells and only four produced 97% of the NPY amacrine cells. Some retinal progenitors did not contribute at all to these two amacrine subtypes. There also is a marked asymmetry in the blastomere origin of DA and NPY amacrine cells. Two retinal progenitors produced significant numbers of NPY but very few DA amacrine cells. This analysis provides evidence that blastomere origin restricts the developmental choices of retinal progenitors.