Ascidians have played a major role in studies to understand the function of cytoplasmic determinants in animal development. Special qualities, including eggs with colored cytoplasmic regions, an invariant cleavage pattern and cell lineage, embryos with low cell numbers, larvae with typical chordate features and only six different tissues, rapid development, and a small genome, combine to make these animals a unique system for studying cytoplasmic determinants. There is evidence for determinants that specify the cleavage pattern; the differentiation of epidermal, endodermal, and muscle cells; and cell movements associated with gastrulation. The muscle determinants appear to be modified in concert with tail and muscle regression in species that have evolved an anural, or tailless, larva. Several lines of evidence suggest that determinants may be localized maternal mRNAs, which encode transcription factors or signal transduction components responsible for initiating differential gene activity. Different approaches and strategies are being used to isolate and characterize the function of these localized maternal mRNAs.