Fertilized eggs of the ascidian, Ciona intestinalis, were prevented from undergoing cytokinesis but not nuclear division by treatment with cytochalasin B. After appropriate times, such cleavage-arrested multinucleate zygotes developed acetylcholinesterase of larval tail muscle and an alkaline phosphatase ordinarily localized in the larval endoderm tissues. Separate histochemical reactions on one of a pair of samples taken from the eggs of single animals provided examples (6134) in which the numbers of cytochalasin-treated embryos displaying the respective reaction product overlapped sufficiently (15-2996) to indicate that some of the zygotes had developed both enzymes in the same uncleaved single cell. With an actual dual-staining technique that can be applied to single cleavage-arrested zygotes, 62% of those developing a strong alkaline phosphatase reaction also had a strong acetylcholinesterase reaction. In other experiments, quantitative measurements of enzyme activity in homogenates of 114 single cleavage-arrested zygotes confirm directly that 18% of the zygotes produce both enzymes. There was no obligatory mutual exclusion of the potential for simultaneous expression of two tissue-specific characteristics that would ordinarily be segregated into different lineages during early cleavages. The cytoplasmic determinants believed responsible for these histotypic expressions can apparently function independently in the same cell.