Suppression of muscle fate by cellular interaction is required for mesenchyme formation during ascidian embryogenesis

Abstract

The tadpole larva of the ascidian Halocynthia roretzi has several hundred mesenchyme cells in its trunk. Mesenchyme cells are exclusively derived from the B8.5 and B7.7 blastomere pairs of the 110-cell embryo. It has been believed that specification of mesenchyme cells is an autonomous process. In the present study, we have demonstrated that presumptive- mesenchyme blastomeres isolated from early 32-cell embryos did not express mesenchyme-specific features, whereas those isolated after the late 64-cell stage developed mesenchyme markers autonomously. Results of experiments involving coisolation and recombination of blastomeres showed that cellular interaction with adjacent presumptive-endoderm blastomeres during the late 32- and early 64-cell stages is required for mesenchyme formation. When such interaction was absent, the presumptive-mesenchyme blastomeres developed into muscle cells. Therefore, a signal from endoderm precursor blastomeres promotes mesenchyme fate, suppressing the muscle fate that is specified by ooplasmic muscle determinants. In Halocynthia, the muscle actin gene was precociously activated in mesenchyme-muscle precursor blastomeres at the 32- cell stage, and the mesenchyme and muscle fates were separated into two daughter blastomeres at the next cleavage. In presumptive-mesenchyme blastomeres at the 64-cell stage, expression of the muscle actin gene was immediately down-regulated by the signal from the neighboring endoderm precursor blastomeres. Thus, mesenchyme formation involves a novel mechanism of fate specification in ascidians, where formation of mesenchyme cells requires cellular interaction that suppresses muscle fate in the mesenchyme precursor blastomeres.